MH_dev_25

Elevated CD3 + and CD8 + tumor-infiltrating immune cells correlate with prolonged survival in glioblastoma patients despite integrated immunosuppressive mechanisms in the tumor microenvironment and at the systemic level . We characterized GBM patients ' tumor and systemic immune contexture with aim to reveal the mechanisms of immunological escape , their impact on patient outcome , and identify targets for immunotherapy . Increased CD3 ( + ) T-cell infiltration was associated with prolonged survival independent of age , P16455 REA promoter methylation and post-operative treatment that implies potential for immunotherapy for GBM . Several mechanisms of escape were identified : within the tumor microenvironment : induced CD8 ( + ) P10747 REA ( - ) Foxp 3 ( + ) Tregs that may tolerize antigen presenting cells , elevated CD73 and P49961 REA ectonucleotidases that suppress T-cell function , and at the systemic level : elevated P22301 REA levels in serum , diminished helper T-cell counts , and upregulated inhibitory P16410 REA .

T cell costimulation : a rational target in the therapeutic armamentarium for autoimmune diseases and transplantation . T cells are central mediators of adaptive immunity . As such , they are involved in both normal immune responses ( e . g . , rejection of a transplanted organ ) and abnormal ones ( e . g . , rheumatoid arthritis ) . T cells require both antigen-specific and costimulatory signals for their full activation . Advances in protein engineering and an increased understanding of the immune response have culminated in the evolution and creation of protein therapeutics that target specific costimulatory molecules . The selective costimulation modulator abatacept ( CTLA - 4Ig ) binds to P33681 REA and P42081 REA , blocking interaction with P10747 REA , and is approved for the treatment of moderate to severe rheumatoid arthritis . DB06681 SUB , currently enrolling phase III trials in renal transplantation , was rationally designed from abatacept to bind with more avidity to P42081 REA , providing the more potent immunosuppressive properties required for immunosuppression in transplantation . This review describes the relevant immunology and summarizes recent clinical findings on these two molecules . Although both inhibit the P10747 REA costimulatory pathway , they are tailored for specific disease states - - abatacept for autoimmune diseases and belatacept for transplantation .

Five-year safety and efficacy of belatacept in renal transplantation . DB06681 SUB is a first-in-class co-stimulation blocker in development for primary maintenance immunosuppression . A Phase II study comparing belatacept with cyclosporine ( DB00091 ) for prevention of acute rejection and protection of renal function in kidney transplant recipients demonstrated similar efficacy and significantly higher measured Q92565 at 1 year for belatacept , but the incidence of posttransplantation lymphoproliferative disorder was higher . Here , we present the results for the extension of this trial , which aimed to assess long-term safety and efficacy of belatacept . Seventy-eight of 102 patients who were receiving belatacept and the 16 of 26 who were receiving DB00091 completed the long-term extension period . Q92565 remained stable in patients who were receiving belatacept for 5 years , and the incidences of death / graft loss or acute rejection were low . The frequencies of serious infections were 16 % for belatacept and 27 % for DB00091 , and neoplasms occurred in 12 % of each group . No patients who were treated with belatacept and one patient who was treated with DB00091 developed posttransplantation lymphoproliferative disorder during the follow-up period . Serious gastrointestinal disorders occurred more frequently with belatacept ( 12 % belatacept versus 8 % DB00091 ) , and serious cardiac disorders occurred more frequently with DB00091 ( 2 % belatacept versus 12 % DB00091 ) . Pharmacokinetic analyses showed consistent exposure to belatacept over time . P42081 REA receptor saturation was higher in patients who were receiving belatacept every 4 weeks ( 74 % ) compared with every 8 weeks ( 56 % ) . In conclusion , this study demonstrated high patient persistence with intravenous belatacept , stable renal function , predictable pharmacokinetics , and good safety with belatacept over 5 years .

Gateways to clinical trials . Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses . The data in the following tables has been retrieved from the Clinical Studies knowledge area of Prous Science Integrity , the drug discovery and development portal , http://integrity.prous.com . This issue focuses on the following selection of drugs : 2F5 , 2G12 , abetimus sodium , ABI - 007 , adalimumab , adefovir dipivoxil , DB05387 , alefacept , altropane , aminolevulinic acid hydrochloride , aminolevulinic acid methyl ester , aminopterin , anakinra , aprinocarsen sodium , atazanavir , atlizumab , atomoxetine hydrochloride ; P33681 REA - 1 vaccine , bevacizumab , DB04851 dicitrate , BMS - 188667 , brasofensine sulfate , bryostatin 1 ; cantuzumab mertansine , Q99698 - 828 , cinacalcet hydrochloride , cipamfylline , creatine , CVT - 3146 ; darbepoetin alfa , DITPA , drotrecogin alfa ( activated ) , duloxetine hydrochloride ; edatrexate , efalizumab , ENMD - 0997 , epoetin , erlosamide , esomeprazole magnesium , etiprednol dicloacetate , etoricoxib , everolimus , ezetimibe ; fampridine , fenretinide , FTY - 720 ; P05019 REA / P17936 REA , IL - 1 cytokine trap , ilodecakin , interferon beta , ISIS - 104838 , ISIS - 2503 , ISIS - 5132 , ivabradine hydrochloride ; lafutidine , lanthanum carbonate , l - DB00125 hydrochloride , DB06681 SUB , lerdelimumab , levetiracetam , levobupivacaine hydrochloride , levosimendan , lopinavir ; melagatran , mibefradil hydrochloride , miglustat , morphine - 6 - glucuronide ; nesiritide ; omalizumab , omapatrilat ; p24 - VLP , parecoxib sodium , peginterferon alfa - 2a , peginterferon alfa - 2b , pegsunercept , pitavastatin calcium , plevitrexed , prasterone , pregabalin , PRO - 2000 , prucalopride ; rapacuronium bromide , rebimastat , RGA - 0853 , rubitecan , ruboxistaurin mesilate hydrate , RWJ - 67657 ; S -16020-2 , sarizotan , SLV - 306 , stiripentol ; DB05809 , tenecteplase , teriparatide , tezacitabine , tipifarnib , trabectedin , troglitazone ; valdecoxib , vardenafil ; Z - 338 , ziconotide .

[ Use of new non-nephrotoxic immunosuppressive drugs in kidney transplantation , especially after ischemia-reperfusion injury ] . Medium - and long-term renal graft survival depends on 4 main factors : the quality of the harvested graft , ischemia-reperfusion injury during harvesting and re-implantation , rejection , and the nephrotoxicity of certain drugs ( especially immunosuppressants ) used in this setting . The most nephrotoxic immunosuppressive drugs are the anticalcineurins ( cyclosporine A and tacrolimus ) , a class discovered in the late 1970s and currently representing a basic component of all immunosuppressive protocols for solid organ graft recipients . The renal tubular and vascular toxicity of anticalcineurins is due to their immunosuppressive mechanism : they block the calcineurin pathway and thereby prevent transmission of the first signal from the T cell receptor to the nucleus , which normally triggers cytokine synthesis , New non-nephrotoxic immunosuppressants are therefore needed , especially for grafts of poor quality or subject to severe ischemia-reperfusion injury . Attention is turning to " old " molecules such as anti-thymocyte globulins , but exciting new immunosuppressants are now appearing . DB00092 is a fusion protein that binds to the immunological synapse-associated molecule P06729 REA , which normally interacts with LFA - 3 . DB06681 SUB , another fusion protein , blocks the T cell second signal CD 28 - P33681 REA . 1 / P33681 REA . 2 . Finally , new chemical agents are being developed , such as sautrasporine , a tyrosine kinase inhibitor , and tofacitinib , a Jak inhibitor .

Serological HLA class I alleles in Senegalese blood donors detected HBs Ag positive . We analysed the HLA class I alleles in 96 blood donors HBs Ag positive compared with 93 healthy control individuals ( HBs negative ) . The most frequent HLA-A , - B , - C alleles found were , A23 ( 33.6 % ) ; A2 ( 25 % ) ; A30 ( 25 % ) ; B8 ( 31.5 % ) ; P33681 REA ( 16.3 % ) ; B58 ( 11.9 % ) ; B35 ( 11.9 % ) ; B49 ( 11.9 % ) ; B53 ( 10.8 % ) ; Cw7 ( Q04695 REA % ) ; Cw3 ( 36.9 % ) ; Cw4 ( 36.9 % ) . Significant differences ( P < 0.001 ) were found between the blood donors and the controls for the following HLA alleles , A1 ; A23 ; B8 and Cw3 . The detection of HBe antigen was positive in 26/84 blood donors . It was observed a significant difference ( P < 0.01 ; odds ratios ( OR )= 6.25 ) between positive and negative HBe antigens blood donors for HLA-A 1 allele .

P29965 REA proinflammatory and profibrotic effects on proximal tubular epithelial cells : role of NF-kappaB and lyn . Chronic allograft nephropathy ( P35658 REA ) is the main cause of renal graft loss , but its pathogenic mechanisms are still unclear . Immune system activation has been suggested as a key event in the development of P35658 REA . P25942 REA is a co-stimulatory protein whose expression is upregulated in proximal tubular epithelial cells ( PTEC ) in acute rejection . This receptor interacts with P29965 REA , expressed by activated T cells . P29965 REA induces the production by PTEC of different proinflammatory cytokines , but very little is known of its profibrotic effects . The aim of this study was to investigate the effect of P25942 REA / P29965 REA interaction on PTEC expression of plasminogen activator inhibitor - 1 ( P05121 REA ) , a powerful profibrotic mediator , and monocyte chemoattractant protein - 1 ( P13500 REA ) , a proinflammatory cytokine , and to investigate the signaling pathways that lead to these effects . Soluble P29965 REA induced a time-dependent increase in both P05121 REA and P13500 REA gene expression and protein production in PTEC . P25942 REA cross-linking on PTEC caused P01375 REA - R-associated factors 2 and 6 membrane translocation . This event led to NF-kappaB activation , through the NF-kappaB-inducing kinase , and to a significant increase in the phosphorylation of lyn , a src-related tyrosine kinase . Lyn , upon phosphorylation , became strictly associated with caveolin - 1 , a scaffolding protein enriched in caveolae . Lyn inhibition did not have any effect on P29965 REA - induced NF-kappaB activation and P13500 REA expression but abolished P05121 REA induction . On the contrary , NF-kappaB inhibition significantly reduced only P13500 REA expression . In conclusion , P29965 REA could play a key role in the pathogenesis of P35658 REA through P05121 REA induction . P29965 REA profibrotic and proinflammatory effects are mediated by different signaling pathways , suggesting that drugs that inhibit inflammation may not be equally effective in reducing fibrosis .

Genetic variants in pre-eclampsia : a meta-analysis . BACKGROUND Pre-eclampsia has a clear familial component , suggesting that the condition may be partly attributable to genetic susceptibility . The search for susceptibility genes has led to a drastic increase in the number of published studies associating genetic factors with pre-eclampsia . However , attempts to replicate these findings have yielded inconsistent results . This meta-analysis assessed the pooled effect of each genetic variant that is reproducibly associated with pre-eclampsia . METHODS Studies that assessed the association between genes and pre-eclampsia were searched in PubMed , Embase and Web of Science . We selected all genetic variants that were significantly associated with pre-eclampsia in an initial study and were subsequently independently reproduced in at least one additional study . All studies that assessed these reproduced variants were then included . The association between genetic variants and pre-eclampsia was calculated at the allele level , and the main measure of effect was a pooled odds ratio in a random-effects model . RESULTS The literature search yielded 2965 articles , of which 542 investigated genetic associations in pre-eclampsia . We identified 22 replicated genetic variants , of which 7 remained significantly associated with pre-eclampsia following meta-analysis . These variants were in or near the following genes : P12821 REA , P16410 REA , F2 , FV , P06858 REA and P05121 REA . CONCLUSIONS This meta-analysis identified seven genetic variants associated with pre-eclampsia . Importantly , many of these variants are also risk factors for developing cardiovascular disease , revealing that pre-eclampsia and cardiovascular disease have shared genetic risk factors . The contribution of the identified genetic variants in the pathogenesis of pre-eclampsia should be the focus of future studies .

Therapeutic factor VIII does not trigger Q15399 REA . 2 and O60603 REA . 6 signalling in vitro . The administration of therapeutic factor VIII ( FVIII ) to patients with haemophilia A induces the development of inhibitory anti-FVIII IgG in a substantial number of patients . For an antigen-specific immune response to develop , antigen-presenting cells ( APCs ) need to mature and procure appropriate co-stimulatory signals to T cells at the time of presentation of the endocytosed antigen . The nature of the danger signals that induce P25054 REA maturation , thus initiating the anti-FVIII immune response , are yet ill-characterized . Contradictory reports on a direct effect of therapeutic FVIII on P25054 REA maturation have been released . Here , we investigated whether FVIII directly triggers O60603 REA ( O60603 REA ) signalling . The capacity of human recombinant FVIII to promote the maturation of a mouse bone marrow macrophage cell line ( BMA ) was investigated by flow cytometry . In parallel , the triggering of Q15399 REA . 2 or O60603 REA . 6 - expressing HEK 293 cells by FVIII was analysed following transfection of the cells with a reporter construct for NFκB activity . In contrast , to zymosan , a known O60603 REA agonist , human recombinant FVIII did not induce the maturation of mouse BMA macrophages , as analysed by the levels of expression of P33681 REA , P42081 REA , P25942 REA and I-Ab at the cell surface . Furthermore , incubation of FVIII with cells expressing O60603 REA paired with Q15399 REA or Q9Y2C9 REA , failed to activate NFκB , whereas NKκB activity was triggered in the presence of zymosan . Our results confirm that FVIII alone is insufficient to trigger the maturation of APCs that is required to initiate an immune response .

DB00278 MEN - coupled Affi-Gel matrix for the purification of thrombin from plasma . Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay . None of the available purification methods easily deals with this subject . The procedure described in the present paper uses a readily available pharmaceutical agent , argatroban , to construct an affinity matrix . DB00278 MEN has a high affinity for thrombin and its thrombin binding is reversible . P00734 REA derived from a Ba ( 2 + ) precipitate of human plasma is used as the starting material . The crude prothrombin can be bulk activated to thrombin using taipan-snake ( Oxyuranus scutellatus ) venom and bound to the argatroban-coupled matrix without further processing steps . The thrombin product eluted from the argatroban matrix is very pure as judged by high specific activity and by electrophoresis . This purification scheme is rapid , yielding purified thrombin within 2 days .

Review : molecular pathology in adult high-grade gliomas : from molecular diagnostics to target therapies . The classification of malignant gliomas is moving from a morphology-based guide to a system built on molecular criteria . The development of a genomic landscape for gliomas and a better understanding of its functional consequences have led to the development of internally consistent molecular classifiers . However , development of a biologically insightful classification to guide therapy is still a work in progress . Response to targeted treatments is based not only on the presence of drugable targets , but rather on the molecular circuitry of the cells . Further , tumours are heterogeneous and change and adapt in response to drugs . Therefore , the challenge of developing molecular classifiers that provide meaningful ways to stratify patients for therapy remains a major challenge for the field . In this review , we examine the potential role of P16455 REA methylation , O75874 REA / 2 mutations , 1p / 19q deletions , aberrant epidermal growth factor receptor and PI3K pathways , abnormal p53 / Rb pathways , cancer stem-cell markers and microRNAs as prognostic and predictive molecular markers in the setting of adult high-grade gliomas and we outline the clinically relevant subtypes of glioblastoma with genomic , transcriptomic and proteomic integrated analyses . Furthermore , we describe how these advances , especially in epidermal growth factor receptor / PI3K / P42345 REA signalling pathway , affect our approaches towards targeted therapy , raising new challenges and identifying new leads .

Concise prediction models of anticancer efficacy of 8 drugs using expression data from 12 selected genes . We developed concise , accurate prediction models of the in vitro activity for 8 anticancer drugs ( DB00544 MEN , DB00515 , DB00305 , DOX , CPT - 11 , SN - 38 , TXL and TXT ) , along with individual clinical responses to DB00544 MEN using expression data of 12 genes . We first performed cDNA microarray analysis and MTT assay of 19 human cancer cell lines to sort out genes which were correlative in expression levels with cytotoxicities of the 8 drugs ; we selected 13 genes with proven functional significance to drug sensitivity from a huge number of potent prediction marker genes . The correlation significance of each was confirmed using expression data quantified by real-time RT-PCR , and finally 12 genes ( P08183 REA , Q9UNQ0 , P10632 REA , P08684 REA , Q12882 REA , P09211 REA , P16455 REA , P15559 REA , P16435 REA , P11388 REA , P07437 REA and P04818 REA ) were selected as more reliable predictors of drug response . Using multiple regression analysis , we fixed 8 prediction formulae which embraced the variable expressions of the 12 genes and arranged them in order , to predict the efficacy of the drugs by referring to the value of Akaike ' s information criterion for each sample . These formulae appeared to accurately predict the in vitro efficacy of the drugs . For the first clinical application model , we fixed prediction formulae for individual clinical response to DB00544 MEN in the same way using 41 clinical samples obtained from 30 gastric cancer patients and found to be of predictive value in terms of survival , time to treatment failure and tumor growth . None of the 12 selected genes alone could predict such clinical responses .

DB06643 MEN - - an emerging treatment for postmenopausal osteoporosis . IMPORTANCE OF THE FIELD : Osteoporosis is a common skeletal disease that is associated with an imbalance in bone remodeling . DB06643 MEN is an investigational fully human monoclonal antibody to receptor activator of NF-kappaB ligand ( O14788 REA ) , a cytokine member of the P01375 REA family that is the principal mediator of osteoclastic bone resorption . AREAS COVERED IN THIS REVIEW : The efficacy and safety of denosumab in the management of postmenopausal osteoporosis is evaluated by reviewing the published literature and presentations at scientific meetings through 2009 . WHAT THE READER WILL GAIN : This review focuses on the data on fracture risk reduction and safety endpoints of denosumab in the treatment of postmenopausal osteoporosis . TAKE HOME MESSAGE : In postmenopausal women with osteoporosis , denosumab ( 60 mg by subcutaneous injection every 6 months ) increased bone mineral density , reduced bone turnover markers , and reduced the risk of vertebral , hip and non-vertebral fractures . DB06643 MEN was well tolerated with a safety profile generally similar to placebo . It is a promising emerging drug for the prevention and treatment of postmenopausal osteoporosis .

Mechanisms of ganglioside inhibition of P25054 REA function . Gangliosides shed by tumor cells exert potent inhibitory effects on cellular immune responses . Here we have studied ganglioside inhibition of P25054 REA function . When human monocytes were preincubated in 50 micro M highly purified ganglioside G ( D1a ) , pulsed with tetanus toxoid ( TT ) , and washed , the expected Ag-induced proliferative response of autologous normal T cells added to these monocytes was inhibited by 81 % . Strikingly , there was also almost complete ( 92 % ) and selective inhibition of the up-regulation of the monocyte costimulatory molecule P33681 REA , while I - P62158 - 1 , LFA - 3 , HLA-DR , and P42081 REA expression were unaffected . Purified LPS-stimulated monocytes that had been preincubated in G ( D1a ) likewise showed inhibition of P33681 REA up-regulation ( 59 % ) as well as down-regulation of P25942 REA ( 54 % ) and impaired release of IL - 12 and P01375 REA ( reduced by 59 and 51 % ) . G ( D1a ) - preincubated human dendritic cells ( DC ) were also affected . They had reduced constitutive expression of P25942 REA ( 33 % ) and P33681 REA ( 61 % ) , but not P42081 REA , and marked inhibition of release of P05231 REA ( 72 % ) , IL - 12 ( 70 % ) , and P01375 REA ( 46 % ) . Even when pulsed with TT , these ganglioside-preincubated DC remained deficient in costimulatory molecule expression and cytokine secretion and were unable to induce a normal T cell proliferative response to TT . Finally , significant inhibition of nuclear localization of NF-kappaB proteins in activated DC suggests that disruption of NF-kappaB activation may be one mechanism contributing to ganglioside interference with P25054 REA expression of costimulatory molecules and cytokine secretion , which , in turn , may diminish antitumor immune responses .

Development of a chimeric anti - P25942 REA monoclonal antibody that synergizes with DB06681 SUB to prolong islet allograft survival . In recent years , reagents have been developed that specifically target signals critical for effective T cell activation and function . Manipulation of the P10747 REA / P33681 REA /8 6 and P25942 REA / CD154 pathways has exhibited extraordinary efficacy , particularly when the pathways are blocked simultaneously . Despite the reported efficacy of anti-CD 154 in rodents and higher models , its future clinical use is uncertain due to reported thromboembolic events in clinical trials . To circumvent this potential complication , we developed and evaluated a chimeric Ab targeting P25942 REA ( Chi 220 , BMS - 224819 ) as an alternative to CD154 . Although Chi 220 blocks CD154 binding , it also possesses partial agonist properties and weak stimulatory potential . The anti - P25942 REA was tested alone and in combination with a rationally designed , high affinity variant of P16410 REA - Ig , DB06681 SUB ( belatacept ) , in a nonhuman primate model of islet transplantation . Although either agent alone only modestly prolonged islet survival ( Chi 220 alone : 14 , 16 , and 84 days ; DB06681 SUB alone : 58 and 60 days ) , their combination ( DB06681 SUB and Chi 220 ) dramatically facilitated long term survival ( 237 , 237 , 220 , > 185 , and 172 days ) . We found that the effects of Chi 220 treatment were not mediated solely through deletion of P11836 - bearing cells and that the combined therapy did not significantly impair established antiviral immunity .

Bayesian analysis and the GUSTO trial . Global Utilization of DB00086 MEN and Tissue P00747 REA Activator in Occluded Arteries .

DB06643 MEN for joints and bones . DB06643 MEN is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 REA ) , a cytokine member of the tumor necrosis factor family . O14788 REA , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 MEN suppresses bone turnover by inhibiting the action of O14788 REA on osteoclasts . DB06643 MENMAX DB06643 MEN reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo .

P00734 REA in normal human cerebrospinal fluid originates from the blood . In spite of the fact that prothrombin is produced by cells within the central nervous system , its presence in the cerebrospinal fluid ( P04141 REA ) has not been investigated . We determined the concentration of prothrombin in P04141 REA with reference to the concentration in plasma in paired samples from 18 " normal " control patients and 4 patients with relapsing-remitting type of multiple sclerosis ( MS ) . The newly developed ELISA was very specific ( no cross-reactivity with thrombin ) and sensitive ( detection limit - - 0.7 ng / ml ) with an imprecision of CV = 8.3 % ( intraseries ) and 7.0 % ( interassay ) . The mean prothrombin concentration in normal P04141 REA was 0.55 mg / l ( CV + / - 33 % , range : 0.28- 0.93 mg / l ) , in normal plasma 121.8 mg / l + / - 21 % , resulting in a mean P04141 REA / plasma concentration quotient ( Q ( Proth ) - - 4.5 x 10 ( - 3 ) ( CV + / - 35 % , range : 2.1- 8.3 x 10 ( - 3 ) ) corresponding to a mean albumin quotient in this group of subjects of Q ( Alb ) = 5.8 x 10 ( - 3 ) . Due to the Q ( Proth ) and the molecular weight of prothrombin ( 72 kDa ) - - similar to that of albumin - - we conclude that prothrombin in normal human P04141 REA originates predominantly ( > 95 % ) from blood . The enzymatic activity in P04141 REA is conserved . Comparable results obtained in MS patients with only few small Q9BWK5 lesions suggest that local chronic inflammatory disease of the central nervous system does not influence prothrombin concentration in the P04141 REA if the blood - P04141 REA barrier function is normal .

P00797 REA inhibition reduces atherosclerotic plaque neovessel formation and regresses advanced atherosclerotic plaques . OBJECTIVE : The interaction between the renin-angiotensin system and toll-like receptors ( TLRs ) in the pathogenesis of advanced atherosclerotic plaques is not well understood . We studied the effects of the renin inhibitor aliskiren on the progression of advanced atherosclerotic plaque in apolipoprotein E-deficient ( ApoE ( - / - ) ) mice with a special focus on plaque neovessel formation . METHODS AND RESULTS : Four-wk-old ApoE ( - / - ) mice were fed a high-fat diet for 8 wks , and the mice were randomly assigned to one of three groups and administered a vehicle , hydralazine , or aliskiren for an additional 12 wks . DB09026 MEN reduced the atherosclerotic plaque area and plaque neovessel density . It increased the plaque collagen and elastin contents , and reduced plasma angiotensin II levels and plaque macrophage infiltration and cathepsin S ( CatS ) protein . DB09026 MEN also decreased the levels of AT1R , gp91phox , O60603 REA , monocyte chemotactic protein - 1 , and CatS mRNAs in the aortic roots . DB01275 had no beneficial vascular effects , although its administration resulted in the same degree of blood pressure reduction as aliskiren . CatS deficiency mimicked the aliskiren-mediated vasculoprotective effect in the ApoE ( - / - ) mice , but aliskiren showed no further benefits in ApoE ( - / - ) CatS ( - / - ) mice . In vitro , O60603 REA silencing reduced CatS expression induced by angiotensin II . Moreover , aliskiren or the inhibition of CatS impaired the endothelial cell angiogenic action in vitro or / and ex vivo . CONCLUSION : P00797 REA inhibition appears to inhibit advanced plaque neovessel formation in ApoE ( - / - ) mice and to decrease the vascular inflammatory action and extracellular matrix degradation , partly by reducing AT1R / O60603 REA - mediated CatS activation and activity , thus regressing advanced atherosclerosis .

P00797 REA angiotensin system modulates P42345 REA pathway through AT2R in HIVAN . P42345 REA ( P42345 REA ) has been reported to contribute to the development of HIV-associated nephropathy ( HIVAN ) . We hypothesized that HIV may be activating renal tissue P42345 REA pathway through renin angiotensin system ( DB01367 ) via Angiotensin Receptor Type II receptor ( AT2R ) . Renal tissues of Vpr transgenic and Tg26 ( HIVAN ) mice displayed enhanced phosphorylation of P42345 REA and p70S6K . DB09026 MEN , a renin inhibitor attenuated phosphorylation of both P42345 REA and p70S6K in renal tissues of HIVAN mice . Interestingly , Angiotensin Receptor Type I ( AT1R ) blockade did not modulate renal tissue phosphorylation of P42345 REA in HIVAN mice ; on the other hand , AT2R blockade attenuated renal tissue phosphorylation of P42345 REA in HIVAN mice . In vitro studies , both renin and Ang II displayed enhanced mouse tubular cell ( P04629 REA ) phosphorylation of p70S6K in a dose dependent manner . HIV / P04629 REA also displayed enhanced phosphorylation of both P42345 REA and p70S6K ; interestingly this effect of HIV was further enhanced by losartan ( an AT1R blocker ) . On the other hand , AT2R blockade attenuated HIV-induced tubular cell phosphorylation of P42345 REA and p70S6K , whereas , AT2R agonist enhanced phosphorylation of P42345 REA and p70S6K . These findings indicate that HIV stimulates P42345 REA pathway in HIVAN through the activation of renin angiotensin system via AT2R .

P49771 REA promotes myeloid dendritic cell differentiation of human hematopoietic progenitor cells : possible application for cancer immunotherapy . Current in vitro culture systems allow the generation of human dendritic progenitor cells ( CFU-DCs ) . The aim of this study was to assess the effect of P49771 REA ( FL ) on the proliferation of human peripheral blood-derived myeloid CFU-DCs and their differentiation into more committed precursor cells ( pDCs ) using in vitro culture systems . Immunomagnetically separated P28906 REA + cells were cultured in serum-free , as well as in serum-containing , liquid suspension cultures to investigate the expansion and / or proliferation / differentiation of CFU-DCs , pDCs , and more mature dendritic cells ( DCs ) . FACS-sorted P28906 REA + Flt 3 + / - cells were cultured in methylcellulose to assay hematopoietic progenitors , including CFU-DCs . In the clonal cell culture supplemented with granulocyte / macrophage ( GM ) colony-stimulating factor ( P04141 REA ) , interleukin - 4 , and tumor necrosis factor alpha , the frequency of CFU-DCs was significantly higher in the P28906 REA + Flt 3 + fraction than in the P28906 REA + Flt 3 - population , thus suggesting functional Flt 3 expression on CFU-DCs . Serum-free suspension culture of P28906 REA + cells revealed the potent effect of FL on the expansion of CFU-DCs in synergy with GM - P04141 REA and thrombopoietin ( P07202 REA ) . In addition , FL strongly induced the maturation of CFU-DCs into functional CD1a + pDCs in serum-containing liquid suspension culture . Moreover , these FL-generated pDCs showed remarkable potential to differentiate into mature DCs with surface Q01151 / P42081 REA expression , which induced a distinct allogeneic T-cell response . These results clearly demonstrate that FL supports not only the proliferation of early hematopoietic progenitor cells , but also the maturation process of committed precursor cells along with the DC-lineage differentiation . Therefore , it is possible to develop a more efficient DC-based cancer immunotherapy using this specific cytokine combination , GM - P04141 REA + P07202 REA + FL in vitro in the near future .

Quantitative analysis predicts the relative therapeutic efficacy of different forms of DB01281 . Modulating the activities of costimulatory molecules controlling immune responses holds considerable promise for immunotherapy . DB01281 ( abatacept ) , a soluble version of the T cell-expressed membrane receptor P16410 REA , is approved for the treatment of rheumatoid arthritis . Like natural P16410 REA molecules , DB01281 ligates P33681 REA - 1 and P33681 REA - 2 on antigen presenting cells , preventing P10747 REA - mediated costimulation of T cells . However , DB01281 can also prevent ligation of P16410 REA , potentially blocking vital inhibitory signals , thereby augmenting immunity . There have been no quantitative analyses of the likely effects of DB01281 on costimulatory interactions at the immunological synapse . We present a mathematical model , based on rigorous biophysical and expression data , for simulating the effects of abatacept and a mutated derivative , DB06681 SUB , on the synaptic interactions of P10747 REA and P16410 REA . The simulations reveal an unexpectedly large window within which P10747 REA , but not P16410 REA , ligation is blocked by DB01281 , perhaps explaining the efficacy of abatacept at the recommended therapeutic dose ( 10mg / kg ) and its relative safety . However , the simulations suggest that the present dosing regimen is close to the maximum theoretically safe dose . The simulations also show that , within the therapeutic window , DB06681 SUB enhances the interaction of P16410 REA with the more potent of its two native ligands , P33681 REA - 1 . They also suggest that P16410 REA ligation by P33681 REA - 1 could , in principle , be enhanced by further decreasing the off-rate of DB01281 for binding to P33681 REA - 2 . Our findings therefore offer molecular explanations for why DB06681 SUB might prove to be more effective than abatacept in a clinical setting , and suggest ways in which its therapeutic efficacy could be further optimised .

T-cell phenotype in protocol renal biopsy from transplant recipients treated with belatacept-mediated co-stimulatory blockade . BACKGROUND : DB06681 SUB is thought to disrupt the interaction between P33681 REA /8 6 and P10747 REA , thus preventing T-cell activation by blocking the co-stimulatory second signal . However , the consequences on the T-cell profile in human renal transplant cases have not been determined . METHODS : In this study , we analysed intra-graft levels of the mRNAs for Treg ( Q9BZS1 ) , cytotoxic CD8 T cells ( P10144 REA ) , Th1 ( INFγ , Tbet ) , Th2 ( P23771 REA ) and Th17 ( RORγt and Q16552 REA ) in protocol biopsies obtained 12 months after renal transplantation in recipients treated with DB06681 SUB or calcineurin inhibitor ( CNI ) . RESULTS : Only the intra-graft abundance of Q9BZS1 mRNA was significantly lower ( P < 0.001 ) in the DB06681 SUB group than the CNI group . Conclusions . These results are in agreement with in vitro data suggesting that P10747 REA is a major co-stimulatory signal of both Tregs development and peripheral homeostasis but contrast with clinical trials showing a better 1 - year graft function and a lower incidence of chronic allograft nephropathy in patients receiving DB06681 SUB than patients treated with CNI . They suggest that immune benefits induced by DB06681 SUB are not mediated by Treg expansion and that Q9BZS1 is not by itself a prognostic marker of long-term graft function in a non-inflammatory context . These results have to be , however , considered as preliminary since the size of our study population is limited .

Maturation of dendritic cells by recombinant human P29965 REA - trimer leads to a homogeneous cell population with enhanced surface marker expression and increased cytokine production . Dendritic cells ( DC ) have been shown to be potent inducers of specific cytotoxic T-cell responses both in vivo and in vitro . Furthermore , exposure to cytokines such as tumour necrosis factor ( P01375 REA ) - alpha or P25942 REA triggering changes DC phenotype and cytokine production and may enhance the T-cell activating capacity of the DC . We studied DC phenotype and cytokine production as well as the T-cell proliferation and cytotoxic T lympocyte ( CTL ) activation induced by DC generated in vitro . In addition , the effect of exposure to recombinant human P29965 REA - trimer ( huCD 40LT ) on these parameters was investigated . Effective differentiation of monocytes derived from freshly isolated peripheral blood mononuclear cells ( PBMC ) was obtained with granulocyte macrophage-colony stimulating factor ( GM - P04141 REA ) and interleukin ( IL ) - 4 . The DC expression of human leucocyte antigen ( HLA ) molecules , P33681 REA , Q01151 , and P42081 REA was markedly enhanced by exposure to huCD 40LT even compared to P01375 REA exposure . Only a moderate cytokine production was observed initially , while P01375 REA addition or P25942 REA triggering , especially , induced enhanced production of P05231 REA and IL - 12 p40 . Surprisingly , comparable induction of T-cell proliferation by a DC allostimulus or through the presentation of PPD , and influenza M1 - peptide specific CTL activity was obtained with nonmaturated ( Q01151 - ) and maturated ( Q01151 + ) DC . In conclusion , a final maturation of monocyte-derived DC through huCD 40LT resulted in a highly homogeneous cell population with enhanced surface marker expression and high production of pro-inflammatory cytokines . In addition , the induction of responses to allo or recall antigens presented by huCD 40LT maturated DC was comparable to the responses obtained with the DC maturated through P01375 REA exposure .

[ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 MEN ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 REA ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC / MS / MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r = 0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC / MS / MS analysis ( r = 0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations .

NSA 9 , a human prothrombin kringle - 2 - derived peptide , acts as an inhibitor of kringle - 2 - induced activation in EOC 2 microglia . In neurodegenerative diseases , such as Alzheimer ' s and Parkinson ' s , microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds . P00734 REA and kringle - 2 increase levels of NO and the mRNA expression of P35228 REA , IL - 1beta , and P01375 REA in microglial cells . In contrast , the human prothrombin kringle - 2 derived peptide NSA 9 inhibits NO release and the production of pro-inflammatory cytokines such as IL - 1beta , P01375 REA , and P05231 REA in LPS-activated EOC 2 microglia . In this study , we investigated the anti-inflammatory effects of NSA 9 in human prothrombin - and kringle - 2 - stimulated EOC 2 microglia . Treatment with 20-100 muM of NSA 9 attenuated both prothrombin - and kringle - 2 - induced microglial activation . NO production induced by MAPKs and NF-kappaB was similarly reduced by inhibitors of P29323 REA ( PD98059 ) , p38 ( SB203580 ) , NF-kappaB ( DB06151 ) , and NSA 9 . These results suggest that NSA 9 acts independently as an inhibitor of microglial activation and that its effects in EOC 2 microglia are not influenced by the presence of kringle - 2 .

Plasmin-induced proteolysis of tenascin-C : modulation by T lymphocyte-derived urokinase-type plasminogen activator and effect on T lymphocyte adhesion , activation , and cell clustering . Proteolysis and remodeling of the extracellular matrix occur physiologically in processes such as tissue morphogenesis and repair and may participate in the regulation of complex cell functions , including proliferation and differentiation . While matrix degradation appears to be relevant to T lymphocyte migration through tissues , little is known about whether degraded matrix affects T lymphocyte function . We have studied the interaction between T lymphocytes and tenascin-C ( TN-C ) , a matrix protein we have previously reported to inhibit T lymphocyte activation , in the context of plasmin-induced degradation . Here we report that plasmin efficiently cleaves TN-C . Peripheral blood T lymphocytes stimulated with phorbol ester , anti - P10747 REA , or anti-CD 3 Ab , induce , within 24 to 48 h , a strong plasminogen-dependent proteolysis of TN-C . We demonstrate that stimulated T lymphocytes activate plasminogen by secreting the urokinase-type plasminogen activator ( u-PA ) . P00747 REA activation by T lymphocyte-derived u-PA occurs efficiently in fluid phase in the absence of cells . We investigate the consequences of plasmin-induced proteolysis on three of the effects of TN-C in relation to lymphocyte functions . Plasmin proteolysis converts TN-C from a nonadhesive into an adhesive substrate for T lymphocytes and abolishes its aggregating activity on PBMC . In contrast , the inhibitory effect of TN-C on T lymphocyte activation remains unaffected . These observations demonstrate that stimulated T lymphocytes induce plasminogen-dependent proteolysis of TN-C by secreting u-PA and suggest that proteolysis of TN-C may represent a mechanism by which to regulate some of its effects on T lymphocyte functions .

DB00203 MEN inhibits calcineurin / Q13469 REA - mediated cyclin A expression in pulmonary artery smooth muscle cells . AIMS : To examine whether calcineurin / NFAT signaling pathway leads to proliferation of pulmonary artery smooth muscle cells ( PASMCs ) by regulating cell cycle proteins and whether the phosphodiesterase - 5 ( O76074 REA ) inhibitor sildenafil affects calcineurin / NFAT-induced cell proliferation . MAIN METHODS : A [ ( 3 ) H ] thymidine incorporation assay was used to examine DNA synthesis ( cell proliferation ) ; cyclin A and Q13469 REA expressions were determined by Western blot . P24941 REA ( P24941 REA ) activity was measured with an in vitro kinase activity assay , and calcineurin and NFAT activity were evaluated using a calcineurin assay kit and a luciferase activity assay , respectively . A chemical inhibitor or siRNA transfection was used to inhibit calcineurin / NFAT signaling pathway . KEY FINDINGS : Serotonin dose-dependently stimulated cyclin A expression in PASMCs . This effect was accompanied by dose-dependent increases in P24941 REA activity and the rate of DNA synthesis . At the same time , PASMCs treated with serotonin showed dose-dependent activation of calcineurin / NFAT signaling pathway . Inhibition of calcineurin activity by cyclosporine A or loss of Q13469 REA protein by siRNA transfection abolished serotonin-induced cyclin A expression and consequent P24941 REA activation and DNA synthesis . We further found that pretreatment of cells with sildenafil suppressed serotonin-triggered activation of calcineurin / Q13469 REA signaling pathway and resultant cyclin A expression , P24941 REA activation and cell proliferation , while the presence of DT - 3 [ a specific protein kinase G ( PKG ) peptide inhibitor ] reversed the effects of sildenafil on PASMCs . SIGNIFICANCE : Our study suggests that enhanced PKG activity suppresses calcineurin / Q13469 REA cascade-mediated cyclin A expression , P24941 REA activation and PASMC proliferation to contribute to the overall effects of sildenafil in the treatment of pulmonary hypertension .

Synergistic proliferative action of insulin-like growth factor I and 17 beta-estradiol in MCF - 7S breast tumor cells . We have analyzed the mechanism by which the combination of insulin-like growth factor I ( P05019 REA ) and 17 beta-estradiol ( E2 ) induces cell cycle progression in MCF - 7S cells . This cell line differs from many other breast cancer-derived cell lines in that E2 ( 1 nM ) does not induce cell cycle progression , whereas the combination of submitogenic concentrations of P05019 REA ( 2 ng / ml ) and E2 does . We find that addition of P05019 REA to MCF - 7S cells leads to a dose-dependent activation of the IGF type I receptor and of the Q96HU1 kinase and P19957 REA - kinase signaling pathways . No synergy of P05019 REA and E2 was detected in the activation of these signaling cascades . In terms of cell cycle-related molecules , we find that P05019 REA dose-dependently raises cyclin D1 levels in serum-starved cells . Subsequent activation of cyclin E / P24941 REA , hyperphosphorylation of P06400 REA , and DNA synthesis are only induced by mitogenic concentrations of P05019 REA ( > or = 20 ng / ml ) . Treatment of the cells with E2 also results in the induction of cyclin D1 , but in the absence of P05019 REA the cells remain arrested in P55008 phase . We conclude that in MCF - 7S cells , the synergistic action of E2 and P05019 REA derives from the ability of both hormones to induce cyclin D1 expression . The action of P05019 REA is required in these cells to induce activity of the cyclin D1 / P11802 REA complex , which triggers progression through the cell cycle .

Mesenchymal stem cells control alloreactive CD8 ( + ) P10747 REA ( - ) T cells . P10747 REA / P33681 REA co-stimulation blockade with belatacept prevents alloreactivity in kidney transplant patients . However , cells lacking P10747 REA are not susceptible to belatacept treatment . As CD8 ( + ) P10747 REA ( - ) T-cells have cytotoxic and pathogenic properties , we investigated whether DB05914 ( O60682 ) are effective in controlling these cells . In mixed lymphocyte reactions ( P08235 REA ) , O60682 and belatacept inhibited peripheral blood mononuclear cell ( PBMC ) proliferation in a dose-dependent manner . O60682 at O60682 / effector cell ratios of 1:160 and 1:2 · 5 reduced proliferation by 38 · 8 and 92 · 2 % , respectively . DB06681 SUB concentrations of 0 · 1 μg / ml and 10 μg / ml suppressed proliferation by 20 · 7 and 80 · 6 % , respectively . Both treatments in combination did not inhibit each other ' s function . Allostimulated CD8 ( + ) P10747 REA ( - ) T cells were able to proliferate and expressed the cytolytic and cytotoxic effector molecules granzyme B , interferon ( IFN ) - γ and tumour necrosis factor ( P01375 REA ) - α . While belatacept did not affect the proliferation of CD8 ( + ) P10747 REA ( - ) T cells , O60682 reduced the percentage of P10747 REA ( - ) T cells in the proliferating CD8 ( + ) T cell fraction by 45 · 9 % ( P = 0 · 009 ) . CD8 ( + ) P10747 REA ( - ) T cells as effector cells in P08235 REA in the presence of P01730 REA ( + ) T cell help gained P10747 REA expression , an effect independent of O60682 . In contrast , allostimulated P10747 REA ( + ) T cells did not lose P10747 REA expression in P08235 REA - O60682 co-culture , suggesting that O60682 control pre-existing P10747 REA ( - ) T cells and not newly induced P10747 REA ( - ) T cells . In conclusion , alloreactive CD8 ( + ) P10747 REA ( - ) T cells that remain unaffected by belatacept treatment are inhibited by O60682 . This study indicates the potential of an O60682 - belatacept combination therapy to control alloreactivity .

Assessment of belatacept-mediated costimulation blockade through evaluation of P33681 REA /8 6 - receptor saturation . BACKGROUND : The selective inhibitor of T-cell costimulation , belatacept , blocks P10747 REA - mediated T-cell activation by binding P33681 REA and P42081 REA on antigen-presenting cells . Understanding the extent to which belatacept binds to its targets in patients may enable correlation of belatacept exposure to receptor saturation as a pharmacodynamic measure of costimulation blockade . METHODS : Flow cytometry-based receptor competition assays were developed to monitor concentration-dependent occupancy of P33681 REA and P42081 REA receptors in whole blood and dendritic cell cultures in vitro . Receptor occupancy was correlated with inhibition of mixed leukocyte reactions and clinical validation was obtained by comparing receptor saturation in whole blood from healthy volunteers and in de novo renal transplant recipients participating in studies comparing cyclosporine and belatacept-based immunosuppression . RESULTS : DB06681 SUB saturated P33681 REA and P42081 REA receptors in whole blood and dendritic cell cultures , although the belatacept concentrations required for P42081 REA - receptor saturation were approximately 10 - fold higher than those required for P33681 REA saturation ( IC50 = 0.102 microg / mL vs . 0.009 microg / mL ) . Primary alloresponses were inhibited at the belatacept concentration required for P42081 REA - receptor saturation , but not at the lower concentration needed to saturate P33681 REA . Whole blood from belatacept-treated patients had significantly lower levels of free P42081 REA receptors versus pretransplant levels , healthy volunteers , or cyclosporine-treated patients . P42081 REA - receptor saturation correlated with belatacept dose / dose frequency and remained consistently more than 80 % . CONCLUSIONS : These results suggest that belatacept-mediated inhibition of alloresponses involved in transplant rejection correlates with P42081 REA saturation , indicating that P42081 REA - receptor occupancy may be a valid pharmacodynamic measure of costimulation blockade and provide the first direct clinical evidence that belatacept binds to one of its targets .

Upregulation of cell-surface-associated plasminogen activation in cultured keratinocytes by interleukin - 1 beta and tumor necrosis factor-alpha . Keratinocytes synthesize and secrete urokinase-type plasminogen activator ( uPA ) which is bound in an autocrine manner to a specific receptor ( uPA-R ) at the keratinocyte surface . P00747 REA that is also bound to specific membrane binding sites is readily activated by uPA-R-bound uPA . Thus , plasmin is provided for proteolysis of pericellular glycoproteins . The expression of uPA and the uPA-R is confined to migrating keratinocytes during epidermal wound healing , rather than to keratinocytes of the normal epidermis . The regulatory factors of uPA / uPA-R expression in keratinocytes remained largely elusive . Proinflammatory cytokines , such as tumor necrosis factor-alpha ( P01375 REA ) or interleukin - 1 beta ( P01584 REA ) , are present in epidermal wounds . We have therefore tested P01584 REA and P01375 REA for their influence on surface-associated plasminogen activation in a human keratinocyte cell line ( HaCaT ) as well as in primary cultures of normal human epidermal keratinocytes . Both cytokines induced the secretion of uPA into the culture supernatants and a concomitant increase in uPA activity as well as in uPA and uPA-R antigen at the cell surface . The increase was preceded by an increase in specific mRNA . The induction was accompanied by an accelerated uPA-dependent and plasmin-mediated detachment of HaCaT cells from the culture substratum . Taken together , the proinflammatory cytokines P01584 REA and P01375 REA induced a coordinated increase in uPA and uPA-R as well as increased pericellular plasmin-mediated proteolysis in human epidermal keratinocytes . This function might be an element of the molecular cell biological events during epidermal wound healing .

Interactions of tumor necrosis factor ( P01375 REA ) and P01375 REA receptor family members in the mouse and human . Ligands of the tumor necrosis factor superfamily ( TNFSF ) ( P41273 REA , APRIL , Q9Y275 REA , P32970 REA , P32971 REA , P29965 REA , EDA 1 , Q92838 REA , P48023 REA , Q9UNG2 , O43557 REA , lymphotoxin alpha , lymphotoxin alphabeta , P23510 REA , O14788 REA , TL1A , P01375 REA , O43508 REA , and P50591 REA ) bind members of the P01375 REA receptor superfamily ( TNFRSF ) . A comprehensive survey of ligand-receptor interactions was performed using a flow cytometry-based assay . All ligands engaged between one and five receptors , whereas most receptors only bound one to three ligands . The receptors O75509 REA , Q969Z4 , Q9NS68 , P08138 REA , and mouse TNFRH 3 did not interact with any of the known TNFSF ligands , suggesting that they either bind other types of ligands , function in a ligand-independent manner , or bind ligands that remain to be identified . The study revealed that ligand-receptor pairs are either cross-reactive between human and mouse ( e . g . Tweak / Fn14 , Q9Y6Q6 REA / O14788 REA ) , strictly species-specific ( Q9Y5U5 REA / Q9UNG2 ) , or partially species-specific ( e . g . OX40 / P23510 REA , P25942 REA / P29965 REA ) . Interestingly , the receptor binding patterns of lymphotoxin alpha and alphabeta are redundant in the human but not in the mouse system . Ligand oligomerization allowed detection of weak interactions , such as that of human P01375 REA with mouse P20333 REA . In addition , mouse APRIL exists as two different splice variants differing by a single amino acid . Although human APRIL does not interact with Q96RJ3 , the shorter variant of mouse APRIL exhibits weak but detectable binding to mouse Q96RJ3 .

High LET ( 56 ) Fe ion irradiation induces tissue-specific changes in DNA methylation in the mouse . DNA methylation is an epigenetic mechanism that drives phenotype and that can be altered by environmental exposures including radiation . The majority of human radiation exposures occur in a relatively low dose range ; however , the biological response to low dose radiation is poorly understood . Based on previous observations , we hypothesized that in vivo changes in DNA methylation would be observed in mice following exposure to doses of high linear energy transfer ( LET ) ( 56 ) Fe ion radiation between 10 and 100 cGy . We evaluated the DNA methylation status of genes for which expression can be regulated by methylation and that play significant roles in radiation responses or carcinogenic processes including apoptosis , metastasis , cell cycle regulation , and DNA repair ( P53355 REA , Q9UI08 , 14.3 . 3 , p16 , P16455 REA , and P17936 REA ) . We also evaluated DNA methylation of repeat elements in the genome that are typically highly methylated . No changes in liver DNA methylation were observed . Although no change in DNA methylation was observed for the repeat elements in the lungs of these same mice , significant changes were observed for the genes of interest as a direct effect and a delayed effect of irradiation 1 , 7 , 30 , and 120 days post exposure . At delayed times , differences in methylation profiles among genes were observed . DNA methylation profiles also significantly differed based on dose , with the lowest dose frequently affecting the largest change . The results of this study are the first to demonstrate in vivo high LET radiation-induced changes in DNA methylation that are tissue and locus specific , and dose and time dependent .

Dopamine agonist-induced hypothermia and disruption of prepulse inhibition : evidence for a role of D3 receptors ? The dopamine D3 / D2 receptor agonists 7 - OH-DPAT , quinpirole , quinelorane , and PD128907 , the mixed dopamine agonist apomorphine , the D2 agonist bromocriptine , and the D1 / D5 agonist SKF 38393 were examined in models of hypothermia and prepulse inhibition ( PPI ) in Wistar rats . As dopamine agonist-induced hypothermia has been proposed as a model of D3 receptor function , and dopamine agonists are known to disrupt PPI , drug potencies to induce hypothermia were established and compared with doses necessary to disrupt PPI . 7 - OH-DPAT , quinpirole , quinelorane , PD128907 , and apomorphine , reduced body temperature and disrupted PPI with a similar rank order of potency ( quinelorane > quinpirole = 7 - OH-DPAT > PD128907 = apomorphine ) . DB01200 MEN and SKF 38393 were ineffective in both models . In a separate study , the dopamine reuptake inhibitors cocaine and GBR 12909 had no effect on PPI . In a final set of studies , the D2 / D3 antagonist raclopride blocked both 7 - OH-DPAT-induced hypothermia and 7 - OH-DPAT-induced PPI disruption . The P08908 REA antagonist WAY 100,135 , and the peripheral D2 - like antagonist domperidone had no effect . These findings suggest that the hypothermia and PPI disruptions seen with some of these dopamine agonists may be mediated by central D3 receptors ; however , only studies using more selective dopamine receptor ligands can definitively rule out effects at the D2 or D4 receptors .

P08908 REA receptor activation : short-term effects on the mRNA expression of the P08908 REA receptor and galanin in the raphe nuclei . Systemic administration of the P08908 REA receptor agonist 8 - OH - 2 - ( di-n-propylamino ) - tetralin ( 8 - OH-DPAT ; 0.3 mg / kg , s . c . ) was used to explore the effects of activation of P08908 REA receptors on expression of mRNA coding for P08908 REA receptor , tryptophan hydroxylase ( P17752 REA ) and galanin in the ascending raphe nuclei . 8 - OH-DPAT increased the hybridization signal of the P08908 REA receptor by 105 % in the dorsal raphe nucleus ( P33681 REA ) 30 min after the injection . No effects were seen at the later time points ( 2-8 h ) . In the median raphe nucleus ( B8 ) and the B9 cell group in the medial lemniscus , 8 - OH-DPAT induced a marked decrease in labeling 30 min after injection . At 8 h following 8 - OH-DPAT injection , the effect had shifted to an increase in P08908 REA receptor labeling by 68 % in the B8 area . Importantly 8 - OH-DPAT had no significant effects on the expression of mRNA coding for P17752 REA and galanin . The results suggest an important and differential mechanism for the regulation of P08908 REA receptor mRNA levels in the dorsal and median raphe nuclei . This regulation may be of importance for the differential control of the activity of the ascending 5 - HT neurons , and hence for mood regulation . The results also indicate a dissociation between the effects mediated by P08908 REA receptor functions and those regulating the coexisting peptide galanin in the dorsal raphe .

Production and characterization of DB06681 SUB , a variant of cytotoxic T-lymphocyte antigen 4 - immunoglobulin , in Pichia pastoris . Blocking the P10747 REA / P33681 REA costimulatory pathway is a promising strategy in the treatment of graft rejection , graft-versus-host disease and autoimmune diseases . DB06681 SUB , a high-affinity variant of cytotoxic T-lymphocyte antigen 4 - immunoglobulin ( DB01281 ) , is a more potent inhibitor of the interaction between P10747 REA and P33681 REA than is DB01281 . In a previous study , DB06681 SUB was produced in a mammalian cell system , which is time-consuming and expensive . To obtain DB06681 SUB more efficiently and cost effectively , we attempted to produce DB06681 SUB using a Pichia pastoris expression system . The gene encoding DB06681 SUB , with an additional 6 - DB00117 tag at the N-terminus , was cloned into the yeast vector pPIC 9K and expressed in the P . pastoris strain GS115 . Under the optimized induction conditions for protein expression ( inoculum density , OD ( 600 ) = 80 ; methanol concentration added daily , 1.0- 3.0 % ; induction time point , 72-96 h ; culture medium pH = 6.0 ) , the yield of purified DB06681 SUB was approximately 30 mg l ( - 1 ) by one-step Ni-agarose affinity chromatography . Q96IV0 F treatment showed the purified DB06681 SUB to be post-translational modified by N-linked glycosylation . In biological function assays , DB06681 SUB expressed in P . pastoris demonstrated specific binding to P33681 REA - 1 / P33681 REA - 2 - positive Raji cells and also suppressed lymphocyte proliferation in a dose-dependent manner . These results suggest that DB06681 SUB produced in P . pastoris is biologically active and will be useful for experimental therapy on immunotherapy for transplant rejection and autoimmune diseases .

What ' s in the pipeline ? New immunosuppressive drugs in transplantation . In the pipeline , there are a number of novel immunosuppressive drugs in preclinical development or in early clinical trials . The major target of new agents are cell-surface molecules important in immune cell interactions ( especially the costimulatory pathway ) , signaling pathways that activate T cells , T-cell proliferation and trafficking and recruitment of immune cells responsible for rejection . The most promising biologic agents include a humanized anti-CD 11a ( anti-LFA 1 ) , humanized anti - P33681 REA . 1 / P33681 REA . 2 , a second-generation DB01281 ( DB06681 SUB ) and a humanized antibody to anti - P08575 REA RB . Inhibitors of T-cell activation and signaling are still in preclinical development . The most interesting inhibitors of T-cell proliferation include inhibitors of the Janus protein tyrosine kinase , P52333 REA , and FK778 , a leflunomide analog . Chemokines play an important role in rejection by virtue of their critical role as regulator of trafficking and activation of lymphocytes . Early trials of FTY 720 , a synthetic small molecule with functional homology to sphingosine - 1 phosphate leading to lymphocyte sequestration , appear very promising ; however , enthusiasm for this drug is mitigated by its potential cardiac side-effects . Antagonists to several chemokine receptors , including P32246 REA , P49682 REA and P51681 REA , have been shown to be effective in experimental transplantation and are likely to be considered for clinical development .

Suppression of IL - 12 production by soluble P29965 REA : evidence for involvement of the Q8TCB0 / 42 mitogen-activated protein kinase pathway . IL - 12 is a key cytokine in skewing immune responses toward Th1 - like reactions . Human monocytes / macrophages produce high amounts of bioactive IL - 12 when a priming signal ( P01579 REA or GM - P04141 REA ) precedes a second signal ( e . g . , LPS ) . We and others have previously shown that preincubation with LPS before this stimulation procedure can efficiently and selectively suppress the production of IL - 12 by human monocytes . In this study , we show that an almost complete suppression of IL - 12 production can also be observed after preincubation of monocytes with costimulatory cell surface molecules that bind to members of the TNFR superfamily ( P29965 REA , O14788 REA ( O14788 REA ) ) . The suppression of IL - 12 was observable on the mRNA and protein levels and was not due to endogenous production of known IL - 12 antagonists ( i . e . , P22301 REA , P05112 REA , and PGE ( 2 ) ) , to an increased number of cells undergoing apoptosis , nor to down-regulation of the P01579 REA or P25942 REA receptor . Cell surface expression of the costimulatory molecules P33681 REA and P42081 REA was not reduced by the preincubation procedure , and only a moderate reduction of P05231 REA production was observed . Several studies have identified signal transduction pathways that are activated by P25942 REA signaling , including activation of mitogen-activated protein kinases . The presence of the extracellular signal-related kinase-specific mitogen-activated protein kinase kinase 1/2- specific inhibitors PD98059 and U0126 abrogated suppression induced by sCD 40 ligand or other second signals . This indicates that activation of extracellular signal-regulated kinase 1/2 contributes to the underlying mechanism of IL - 12 suppression . This mechanism may be relevant in other inflammatory responses and may help to develop therapeutic strategies in Th1 - mediated diseases .

Aging impairs induction of cyclin-dependent kinases and down-regulation of p27 in mouse P01730 REA ( + ) cells . To define the link between the early activation defects and the impaired proliferation response of cells from old mice , we characterized the influence of age on expression and activity of proteins that participate in cell-cycle regulation . We found that aging led to significant declines in the ability of mouse P01730 REA ( + ) T cells to respond to CD3 and P10747 REA stimuli by induction of the cyclin-dependent kinases P24941 REA , P11802 REA , and Q00534 REA , whether the defect was assessed by protein level or functional activity . Induction of P24941 REA activity was also impaired in cells from old mice that were activated with PMA plus ionomycin , stimuli that bypass the TCR / CD3 complex , or by CD3 / P10747 REA in the presence of P60568 REA , indicating that the age-related changes lie , at least in part , downstream of the enzymes activated by these stimuli . We also noted an impairment in the ability of P01730 REA ( + ) cells from old mice to down-regulate the CDK inhibitor p27 after activation , but we found no change in induction of P38936 REA , an inhibitor of CDK that may also play other roles in cell-cycle control . Altered CDK activation is likely to mediate the age-related decline in T cell proliferation to polyclonal stimulation .

Agonism at P41595 REA receptors is not a class effect of the ergolines . Restrictive cardiac valvulopathies observed in Parkinson patients treated with the ergoline dopamine agonist pergolide have recently been associated with the agonist efficacy of the drug at 5 - hydroxytryptamine 2B ( P41595 REA ) receptors . To evaluate whether agonism at P41595 REA receptors is a phenomenon of the class of the ergolines , we studied P41595 REA receptor-mediated relaxation in porcine pulmonary arteries to five ergolines which are used as antiparkinsonian drugs . DB01186 and cabergoline were potent full agonists in this tissue ( pEC 50 8.42 and 8.72 ) . DB01200 MEN acted as a partial agonist ( pEC 50 6.86 ) . Lisuride and terguride , however , failed to relax the arteries but potently antagonized 5 - HT-induced relaxation ( pKB 10.32 and 8.49 ) . Thus , agonism at P41595 REA receptors seems not to be a class effect of the ergolines .

DB06681 SUB : a new era of immunosuppression ? Q8N1N2 T-cell activation in alloimmunity requires the engagement of several costimulatory molecules . P16410 REA - Ig and its commercially available fusion proteins , belatacept and abatacept , are used to block P33681 REA /8 6 and promote T-cell tolerance . DB06681 SUB , a higher binding affinity molecule , is currently approved for clinical use in renal transplantation . The results of two Phase III clinical trials showed a similar patient / graft survival , with better renal function at a 3 - year follow-up compared with conventional immunosuppression . There was a higher risk of early rejection and post-transplant lymphoproliferative disorder , especially with EBV-negative patients receiving kidneys from EBV-positive donors . DB06681 SUB - treated groups had a better cardiovascular and metabolic profile . The authors review both preclinical and human studies of P16410 REA - Igs .

Chronic induction . What ' s new in the pipeline . Induction therapy with biological agents was introduced the in the 1970s and the rationale , concepts and approach have remained almost unchanged for 30 years . However , the novel biological agents being developed for induction therapy are being designed for chronic rather than short-term therapy with several objectives : reduce dependence on toxic and nephrotoxic agents , improve outcome and ultimately facilitate the emergence of tolerance . The biological agents include efalizumab , a humanized anti-CD 11a ( anti-LFA 1 ) , anti - CD154 , anti - P25942 REA , a number of agents targeting P40933 REA and its receptor , and costimulation blockade with humanized antibodies to P33681 REA / P42081 REA and the fusion receptor protein DB06681 SUB , a second generation DB01281 . The past decade has witnessed an unprecedented number of small molecules / oral drugs that have been developed and approved for renal transplantation ; the next decade , however , may witness the emergence of a new class of biological induction agents that may displace some of the currently used drugs .

Granulocyte-macrophage colony stimulating factor up-regulates P32246 REA in human neutrophils . Neutrophils ( polymorphonuclear leukocytes ; PMN ) are phagocytic cells instrumental in the clearance of infectious pathogens . Human PMN are commonly thought to respond primarily to chemokines from the CXC family . However , recent findings suggest that under specific cytokine activation conditions , PMN can also respond to some CC chemokines . In this study , the effect of GM - P04141 REA , a well-characterized PMN priming and maturation factor , on CC-chemokine receptor ( CCR ) expression in PMN was investigated . Constitutive expression of P32246 REA and P51677 REA mRNA in PMN was detected by ribonuclease protection assay . Following incubation of PMN with GM - P04141 REA ( 0.01- 10 ng / ml ; 6 h ) P32246 REA mRNA expression was rapidly ( approximately 1 h ) up-regulated . In contrast , no significant induction of P41597 REA , P51677 REA , CCR 4 , or P51681 REA mRNA was observed . P32246 REA protein was also up-regulated by GM - P04141 REA stimulation . GM - P04141 REA - induced up-regulation of P32246 REA showed functional consequences because GM - P04141 REA - treated PMN , but not control cells , responded to the CC chemokines macrophage inflammatory protein - 1alpha , monocyte chemoattractant protein - 3 , and RANTES in assays of chemotactic migration and intracellular calcium mobilization . These results suggest that PMN activated by the proinflammatory cytokine GM - P04141 REA can change their receptor expression pattern and become responsive to CC chemokines .

Reactive oxygen species-dependent P01375 REA converting enzyme activation through stimulation of P41595 REA and alpha 1D autoreceptors in neuronal cells . A major determinant of neuronal homeostasis is the proper integration of cell signaling pathways recruited by a variety of neuronal and non-neuronal factors . By taking advantage of a neuroectodermal cell line ( 1C11 ) endowed with the capacity to differentiate into serotonergic ( 1C115 - HT ) or noradrenergic ( 1C11NE ) neurons , we identified serotonin ( 5 - hydroxytryptamine , 5 - HT ) - and norepinephrine ( NE ) - dependent signaling cascades possibly involved in neuronal functions . First , we establish that P41595 REA receptors and 1D adrenoceptors are functionally coupled to reactive oxygen species ( ROS ) synthesis through NADPH oxidase activation in 1C115 - HT and 1C11NE cells . This observation constitutes the prime evidence that bioaminergic autoreceptors take part in the control of the cellular redox equilibrium in a neuronal context . Second , our data identify P78536 REA ( P01375 REA - Converting Enzyme ) , a member of a disintegrin and metalloproteinase ( ADAM ) family , as a downstream target of the P41595 REA and 1D receptor-NADPH oxidase signaling pathways . Upon P41595 REA or 1D receptor stimulation , ROS fully govern P01375 REA - shedding in the surrounding milieu of 1C115 - HT or 1C11NE cells . Third , P41595 REA and 1Dreceptor couplings to the NADPH oxidase - P78536 REA cascade are strictly restricted to 1C11 - derived progenies that have implemented a complete serotonergic or noradrenergic phenotype . Overall , these observations suggest that P41595 REA and 1D autoreceptors may play a role in the maintenance of neuron - and neurotransmitter-associated functions . Eventually , our study may have implications regarding the origin of oxidative stress as well as up-regulated expression of proinflammatory cytokines in neurodegenerative disorders , which may relate to the deviation of normal signaling pathways .

Rational development of DB06681 SUB ( belatacept ) , a high-affinity variant of P16410 REA - Ig with potent immunosuppressive properties . Current success in organ transplantation is dependent upon the use of calcineurin-inhibitor-based immunosuppressive regimens . Unfortunately , current immunotherapy targets molecules with ubiquitous expression resulting in devastating non-immune side effects . T-cell costimulation has been identified as a new potential immunosuppressive target . The best characterized pathway includes P10747 REA , its homologue P16410 REA and their ligands P33681 REA and P42081 REA . While an immunoglobulin fusion protein construct of P16410 REA suppressed rejection in rodents , it lacked efficacy in primate transplant models . In an attempt to increase the biologic potency of the parent molecule a novel , modified version of P16410 REA - Ig , DB06681 SUB ( belatacept ) , was constructed . Two amino acid substitutions ( L104E and A29Y ) gave rise to slower dissociation rates for both P42081 REA and P33681 REA . The increased avidity resulted in a 10 - fold increase in potency in vitro and significant prolongation of renal allograft survival in a pre-clinical primate model . The use of immunoselective biologics may provide effective maintenance immunosuppression while avoiding the collateral toxicities associated with conventional immunsuppressants .

Advances in immunosuppression for kidney transplantation : new strategies for preserving kidney function and reducing cardiovascular risk . The development of new immunosuppressants for renal transplantation is aimed not only at improving & nbsp ; short-term outcomes , but also at achieving better safety , cardiovascular , and metabolic profiles and at decreasing & nbsp ; nephrotoxicity . DB06681 SUB is a fusion protein that inhibits T cell activation by binding to P33681 REA and P42081 REA antigens . Clinical trials , particularly the BENEFIT and BENEFIT-EXT studies , have shown that belatacept preserves function and structure in renal grafts . The effects of belatacept provide long-term , sustained results , and the safety and efficacy of this drug have been demonstrated in cases of renal transplantation from expanded criteria donors . Compared to calcineurin inhibitors , belatacept is associated with a lower incidence of chronic allograft nephropathy and a more favourable cardiovascular and metabolic profile .

Novel immunosuppression : small molecules and biologics . Kidney transplantation today has excellent short-term outcomes that have paralleled the use of new immunosuppressive agents introduced in the 1990s . In addition to reducing acute rejection , the goals for developing new agents is to improve long-term outcome , minimize nephrotoxicity , and reduce infectious , cardiovascular , and malignancy-related complications . Novel small molecules and biological agents currently in clinical development may help to minimize the use of calcineurin inhibitors and steroids . These small molecules include FTY 720 , a sphingosine phosphate-receptor modulator , FK778 , an inhibitor of pyrimidine synthesis , CP - 690550 , a P52333 REA inhibitor , and AEB - 071 , a protein kinase C inhibitor . The biological agents include drugs targeting interleukin - 15 , anti - P25942 REA , belatacept ( DB06681 SUB ) , a second-generation CTLY 4Ig that blocks the interaction between P33681 REA /8 6 and P10747 REA costimulatory pathways , and efalizumab , a humanized anti-LFA 1 monoclonal antibody . These new agents currently in preclinical and clinical trials appear promising and may represent the emergence of novel immunosuppressive agents that can deliver immunosuppression without long-term toxicity .

DB06681 SUB in clinical and experimental transplantation - progress and promise . DB06681 SUB is a fusion protein composed of the Fc fragment of a human IgG ( 1 ) immunoglobulin linked to the extracellular domain of cytotoxic T-lymphocyte-associated antigen 4 ( P16410 REA ) . P16410 REA is a molecule crucial for T-cell costimulation , selectively blocking the process of T-cell activation . DB06681 SUB binds surface costimulatory ligands ( P33681 REA and P42081 REA ) of antigen-presenting cells . Studies on nonhuman primates , as well as phase II and III clinical trials are here reviewed . DB06681 SUB is a promising therapy in organ transplantation and in the future can be used to induce tolerance .

Cell cycle arrest induced by engagement of Q7Z7D3 on Epstein-Barr virus-positive B-cell lymphoma cell lines . Q7Z7D3 is a recently discovered P33681 REA family member that has inhibitory effects on T-cell immunity . However , the reverse signalling mechanism of the Q7Z7D3 - expressing cells remains unclear . Previous work has shown that Q7Z7D3 expression was enhanced on B cells following Epstein-Barr virus ( EBV ) infection , and engagement of cell-surface-expressed Q7Z7D3 induces cell death of EBV-transformed B cells . Here we found that Q7Z7D3 was constitutively expressed on EBV-positive lymphoma cells , Raji and IM - 9 cells , but was not expressed on EBV-negative lymphoma cells ( Ramos ) . Engagement of Q7Z7D3 significantly reduced cell growth of Raji and IM - 9 cells and resulted in cell cycle arrest at G0 - P55008 phase in a dose - and time-dependent manner . To clarify the mechanism of cell cycle arrest via activation of Q7Z7D3 , cell cycle regulatory factors were examined by reverse transcription-polymerase chain reaction and immunoblotting . We found that Q7Z7D3 triggered down-regulation of P11802 REA / 6 and up-regulation of P38936 REA expression at both protein and RNA levels . Furthermore , P24941 REA and cyclin E / D expression was down-regulated by Q7Z7D3 triggering . Additionally , the down-regulation of phospho-AKT and phospho-cyclin E were clearly detected in Q7Z7D3 - activated Raji cells , but the phosphorylation of p53 was constitutively maintained . These results indicate that Q7Z7D3 - mediated signalling on EBV-positive B-cell lymphoma cells modulates the cell cycle through down-regulation of the AKT pathway . Consequently , Q7Z7D3 may be a new potential target for use in EBV-positive lymphoma therapy .

Costimulation blockade in autoimmunity and transplantation . Signaling through the costimulation receptors is a critical pathway in the regulation of T-cell activation . The selective costimulation inhibitor abatacept ( cytotoxic T lymphocyte-associated antigen 4 - Ig ) binds to P33681 REA and P42081 REA on antigen-presenting cells , blocking interaction with P10747 REA on T cells , and is approved for the treatment of moderate to severe rheumatoid arthritis . DB06681 SUB ( DB06681 SUB ) , currently enrolling phase III trials in renal transplantation , was rationally designed from abatacept to bind with more avidity to P42081 REA , providing the more potent immunosuppressive properties required for immunosuppression in transplantation . This review describes the relevant preclinical studies and summarizes recent clinical findings on these 2 molecules in autoimmune diseases and organ transplantation . Although both inhibit the P10747 REA costimulatory pathway , they are tailored for specific disease states - - abatacept for autoimmune diseases and belatacept for transplantation .

Introduction to the use of belatacept : a fusion protein for the prevention of posttransplant kidney rejection . The development of new immunosuppressive drugs for kidney transplantation resulted both in better short-term outcomes and in decreased metabolic , cardiovascular , and nephrotoxicity risk . DB06681 SUB belongs to a new class of immunosuppressive drugs that selectively inhibits T-cell activation by preventing P10747 REA activation and by binding its ligands P33681 REA - 1 and P33681 REA - 2 . The result is an inactivation of costimulatory pathways . A comparative analysis of the BENEFIT and BENEFIT-EXT datasets showed belatacept regimens resulted in better cardiovascular and metabolic risk profiles than did cyclosporin A ( DB00091 ) regimens : belatacept likewise outperformed DB00091 in terms of lower blood pressure and serum lipids and less new onset diabetes after transplantation . About 20 % of belatacept-treated patients developed adverse effects which included anemia , pyrexia , neutropenia , diarrhea , urinary tract infection , headache , and peripheral edema . At present , belatacept does not seem to predispose patients to a higher rate of infection than DB00091 maintenance immunosuppression . The risk of posttransplant lymphoproliferative diseases was higher in Epstein-Barr virus ( EBV ) - seronegative patients than in EBV-seropositive patients , but the risk may be reduced by use of a less intensive regimen and avoidance of EBV-negative patients and of patients whose pretransplant EBV serology is unknown . DB06681 SUB provides a new option for immunosuppressive therapy in kidney transplantation , but needs further evaluation in terms of the late effects that may derive from prolonged blockage of the costimulatory system and the induction of tolerance status .

P04818 REA genotype-directed chemotherapy for patients with gastric and gastroesophageal junction cancers . BACKGROUND : Retrospective studies indicate associations between TSER ( thymidylate synthase enhancer region ) genotypes and clinical outcomes in patients receiving DB00544 MEN based chemotherapy , but well-controlled prospective validation has been lacking . METHODS : In this phase II study ( NCT 00515216 registered through ClinicalTrials.gov , http://clinicaltrials.gov/show/NCT00515216 ) , patients with " good risk " TSER genotypes ( at least one TSER * 2 allele ) were treated with FOLFOX chemotherapy to determine whether prospective patient selection can improve overall response rates ( ORR ) in patients with gastric and gastroesophageal junction ( GEJ ) cancers , compared with historical outcomes in unselected patients ( estimated 43 % ) . RESULTS : The ORR in genotype-selected patients was Q04695 REA % ( 9 partial responses out of 23 evaluable patients , 95 % CI , 22.2 to 59.2 ) , not achieving the primary objective of improving ORR . An encouraging disease control rate ( DCR , consisting of partial responses and stable diseases ) of 95.7 % was noted and patients with homozygous TSER * 2 genotype showed better tumor response . CONCLUSIONS : In this first prospective , multi-institutional study in patients with gastric or GEJ cancers , selecting patients with at least one TSER * 2 allele did not improve the ORR but led to an encouraging DCR . Further studies are needed to investigate the utility of selecting patients homozygous for the TSER * 2 allele and additional genomic markers in improving clinical outcomes for patients with gastric and GEJ cancers . TRIAL REGISTRATION : ClinicalTrials.gov NCT 00515216 .

Challenges and opportunities in targeting the costimulation pathway in solid organ transplantation . Signaling through the costimulatory pathway is critical in the regulation of T cell activation . DB01281 , a selective costimulatory antagonist FDA approved for the treatment of moderate to severe rheumatoid arthritis , binds to P33681 REA and P42081 REA on antigen presenting cells , blocking the interaction with P10747 REA on T cells . DB06681 SUB , a second generation P16410 REA - Ig with 2 amino acid substitutions , has shown considerable promise in clinical transplantation as part of a maintenance immunosuppression regimen . This review will summarize the role of costimulation in T cell activation , detail the development of costimulation antagonists and highlight the pertinent clinical trials completed and ongoing utilizing belatacept as part of an immunosuppressive regimen in organ transplantation .

DB06681 SUB and sirolimus prolong nonhuman primate islet allograft survival : adverse consequences of concomitant alefacept therapy . Calcineurin inhibitors ( CNI ) and steroids are known to promote insulin resistance , and their avoidance after islet transplantation is preferred from a metabolic standpoint . DB06681 SUB , a P33681 REA - specific mediator of costimulation blockade ( CoB ) , is clinically indicated as a CNI alternative in renal transplantation , and we have endeavored to develop a clinically translatable , belatacept-based regimen that could obviate the need for both CNIs and steroids . Based on the known synergy between CoB and P42345 REA inhibition , we studied rhesus monkeys undergoing MHC-mismatched islet allotransplants treated with belatacept and the P42345 REA inhibitor , sirolimus . To extend prior work on CoB-resistant rejection , some animals also received P06729 REA blockade with alefacept ( P19256 REA - Ig ) . Nine rhesus macaques were rendered diabetic with streptozotocin and underwent islet allotransplantation . All received belatacept and sirolimus ; six also received alefacept . DB06681 SUB and sirolimus significantly prolonged rejection-free graft survival ( median 225 days compared to 8 days in controls receiving basiliximab and sirolimus ; p = 0.022 ) . The addition of alefacept provided no additional survival benefit , but was associated with Cytomegalovirus reactivation in four of six animals . No recipients produced donor-specific alloantibodies . The combination of belatacept and sirolimus successfully prevents islet allograft survival in rhesus monkeys , but induction with alefacept provides no survival benefit and increases the risk of viral reactivation .

DB01197 MEN and lisinopril suppress production of interleukin - 12 by human peripheral blood mononuclear cells . Angiotensin converting enzyme ( P12821 REA ) inhibitors have immunomodulatory functions and can suppress a number of proinflammatory , monocyte / macrophage-derived cytokines . Interleukin - 12 is a cytokine produced primarily by monocytes and macrophages , which plays an essential role in cell mediated immunity and stimulates the development of T helper type 1 immune responses . In this study , we investigated the ability of P12821 REA inhibitors , captopril and lisinopril , to suppress IL - 12 production by human peripheral blood mononuclear cells ( PBMC ) . We show that both P12821 REA inhibitors significantly inhibit production of IL - 12 by PBMC stimulated with bacterial lipopolysaccharide ( LPS ) or Staphylococcus aureus Cowan ( Q96PN6 ) . Although both P12821 REA inhibitors also suppressed P01579 REA production by human anti-CD 3 / anti - P10747 REA - stimulated T-cells , the addition of exogenous P01579 REA to the PBMC stimulation medium does not abrogate the ability of P12821 REA inhibitors to suppress IL - 12 production . Inhibition of IL - 12 was not associated with inhibition of IL - 1beta , but correlated with the suppression of P12821 REA . Therefore , suppression of IL - 12 may contribute to the immunomodulatory effect of P12821 REA inhibitors and may be responsible for the beneficial effect of captopril and other P12821 REA inhibitors in inflammatory or autoimmune conditions in which IL - 12 is involved .

Plasmid DNA encoding human carcinoembryonic antigen ( P06731 REA ) adsorbed onto cationic microparticles induces protective immunity against colon cancer in P06731 REA - transgenic mice . A carcinoembryonic antigen ( P06731 REA ) - based DNA vaccine , adsorbed onto cationic microparticles of poly ( DL-lactide-co-glycolide ) ( P00747 REA ) induced tumor-protective immunity against a lethal challenge of MC38 - P06731 REA colon carcinoma cells in P06731 REA - transgenic mice that was more potent than that of the corresponding naked DNA vaccine . Boosting with a plasmid encoding murine GM - P04141 REA increased the vaccine ' s efficacy leading to a complete rejection of tumor cells in 50 % of mice . This effect was due to activation of MHC class I-restricted CD8 ( + ) T cells coupled with an increased secretion of proinflammatory cytokines P01579 REA , P01375 REA and P60568 REA . Also , specific activation of dendritic cells was indicated by a two-three-fold upregulation of their costimulatory P33681 REA and MHC class II molecules . This approach may be a promising new strategy for the rational design of cancer vaccines for future clinical applications .

Prolonged , low-dose anti-thymocyte globulin , combined with P16410 REA - Ig , promotes engraftment in a stringent transplant model . BACKGROUND : Despite significant nephrotoxicity , calcineurin inhibitors ( CNIs ) remain the cornerstone of immunosuppression in solid organ transplantation . We , along with others , have reported tolerogenic properties of anti-thymocyte globulin ( ATG , Thymoglobulin ® ) , evinced by its ability both to spare Tregs from depletion in vivo and , when administered at low , non-depleting doses , to expand Tregs ex vivo . Clinical trials investigating P33681 REA / P10747 REA blockade ( DB06681 SUB , DB06681 SUB ) in kidney transplant recipients have proven that the replacement of toxic CNI use is feasible in selected populations . METHODS : Rabbit polyclonal anti-murine thymocyte globulin ( mATG ) was administered as induction and / or prolonged , low-dose therapy , in combination with P16410 REA - Ig , in a stringent , fully MHC-mismatched murine skin transplant model to assess graft survival and mechanisms of action . RESULTS : Prolonged , low-dose mATG , combined with P16410 REA - Ig , effectively promotes engraftment in a stringent transplant model . Our data demonstrate that mATG achieves graft acceptance primarily by promoting Tregs , while P16410 REA - Ig enhances mATG function by limiting activation of the effector T cell pool in the early stages of treatment , and by inhibiting production of anti-rabbit antibodies in the maintenance phase , thereby promoting regulation of alloreactivity . CONCLUSION : These data provide the rationale for development of novel , CNI-free clinical protocols in human transplant recipients .

DB01197 MEN reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 REA ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 REA ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 REA inhibitor captopril reduces plasma P05121 REA inhibitor activity , we measured changes in plasma P05121 REA activity ( IU / ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng / ml ) , and serum P12821 REA activity ( IU / L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 REA activity decreased significantly , from 14.0 + / - 0.8 to 11.5 + / - 1.2 IU / L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 REA activity and t-PA antigen also decreased significantly-from 11.9 + / - 2.8 to 5.5 + / - 2.2 IU / ml ( p < 0.02 ) and from 9.9 + / - 1.0 to 7.5 + / - 0.9 ng / ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 REA activity , P05121 REA activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity .

Viral-induced P10747 REA loss evokes costimulation independent alloimmunity . BACKGROUND : DB06681 SUB , a P33681 REA - specific fusion protein , blocks P10747 REA - P33681 REA costimulation and prevents kidney allograft rejection . However , it is ineffective in a sizable minority of patients . Although T-cell receptor and P10747 REA engagement are known to initiate T-cell activation , many human antigen-experienced T-cells lose P10747 REA , and can be activated independent of P10747 REA signals . We posit that these cells are central drivers of costimulation blockade resistant rejection ( CoBRR ) and propose that CoBRR might relate to an accumulation of P10747 REA ( - ) T-cells resulting from viral antigen exposure . MATERIALS AND METHODS : We infected C57BL / 6 mice with polyomavirus ( a BK virus analog ) , murine cytomegalovirus ( a human cytomegalovirus analog ) , and gammaherpesvirus ( HV68 ; an Epstein-Barr virus analog ) and assessed for P10747 REA expression relative to mock infection controls . We then used mixed lymphocyte reaction ( P08235 REA ) assays to assess the alloreactive response of these mice against major histocompatibility complex-mismatched cells . RESULTS : We demonstrated that infection with polyomavirus , murine CMV , and HV68 can induce P10747 REA downregulation in mice . We showed that these analogs of clinically relevant human viruses enable lymphocytes from infected mice to launch an anamnestic , costimulation blockade resistant , alloreactive response against major histocompatibility complex-mismatched cells without prior alloantigen exposure . Further analysis revealed that gammherpesvirus-induced oligoclonal T-cell expansion is required for the increased alloreactivity . CONCLUSIONS : Virus exposure results in reduced T-cell expression of P10747 REA , the target of costimulation blockade therapy . These viruses also contribute to increased alloreactivity . Thus , P10747 REA downregulation after viral infection may play a seminal role in driving CoBRR .

Co-signals in organ transplantation . PURPOSE OF REVIEW : The nonimmune effects of currently used immunosuppressive drugs result in a high incidence of late graft loss due to nephrotoxicity and death . As an immune-specific alternative to conventional immunosuppressants , new biotechnology tools can be used to block the costimulation signal of T-cell activation . RECENT FINDINGS : Many experimental studies , particularly preclinical studies in nonhuman primates , have focused on blocking ' classical ' P33681 REA / P10747 REA and P25942 REA / P29965 REA pathways , which are critical in primary T-cell activation , but also on new P33681 REA / P10747 REA and P01375 REA / P01375 REA - R pathways families of costimulatory molecules that can deliver positive or negative costimulation signals to regulate the alloimmune response . SUMMARY : DB06681 SUB is a new fusion protein derived from P16410 REA - Ig that can be used to prevent acute rejection in renal transplantation instead of calcineurin inhibitors . DB06681 SUB can also prevent acute rejection efficiently in humans and , more interestingly , can improve renal function and cardiovascular risk factors in this population .

The macrophage mannose receptor induces Q16552 REA in response to Candida albicans . The cytokine Q16552 REA controls neutrophil-mediated inflammatory responses . The pattern recognition receptor ( s ) that induce Th17 responses during infection , in the absence of artificial mitogenic stimulation with anti-CD 3 / anti - P10747 REA antibodies , remain obscure . We investigated the innate immune receptors and pathogen-associated molecular patterns involved in triggering Th17 responses during pathogen-specific host defense . The prototypic fungal pathogen Candida albicans was found to induce Q16552 REA more potently than Gram-negative bacteria . Candida mannan , but not zymosan , beta-glucans , Toll-like receptor ( TLR ) agonists , or the Q9HC29 ligand P16444 REA , induced Q16552 REA production in the absence of anti-CD 3 / anti - P10747 REA antibodies . Candida-induced Q16552 REA response was dependent on antigen-presenting cells and the macrophage mannose receptor ( MR ) , demonstrating that Candida mannan is not simply a mitogenic stimulus . The O60603 REA / dectin - 1 pathway , but not O00206 REA or Q9HC29 , amplified MR-induced Q16552 REA production . This study identifies the specific pattern recognition receptors that trigger the Th17 response induced by a human pathogen in the absence of mitogenic stimulation .