Summary Memory B cells play a pivotal role in alloreactivity kidney transplantation. Follicular T helper (Tfh) an important the differentiation of into immunoglobulin-producing plasmablasts [through interleukin (IL)-21]. It is unclear to what extent this cell subset regulates humoral transplant patients, therefore we investigated absolute numbers and function peripheral Tfh (CD4POSCXCR5POS cells) patients before after In addition, studied their relationship with presence donor-specific...

Belatacept, an inhibitor of the CD28-CD80/86 costimulatory pathway, allows for calcineurin-inhibitor free immunosuppressive therapy in kidney transplantation but is associated with a higher acute rejection risk than ciclosporin. Thus far, no biomarker belatacept-resistant has been validated. In this randomized-controlled trial, rate was compared between belatacept- and tacrolimus-treated patients immunological biomarkers were investigated.Forty transplant recipients 1:1 randomized to...

The Jak inhibitor CP-690,550 inhibits alloreactivity and is currently being investigated for prevention of allograft rejection after transplantation. In this study, we examined the effect on IL-2-mediated Jak/STAT5 phosphorylation by CD4(+)CD25(bright)FoxP3(+)CD127(-/low) T cells (Treg) CD4(+)CD25(neg) effector (Teff) in kidney transplant (KTx) patients. Phosphospecific flow cytometry was used to study IL-2-induced intracellular STAT5-phosphorylation. STAT5 (P-STAT5) both Treg Teff, which...

The calcineurin inhibitor, tacrolimus (TAC), inhibits the protein phosphatase activity of calcineurin, leading to suppression nuclear translocation NFAT and inhibition T cell activation. Apart from also transcription factor NF-κB plays a key functional role in Therefore, blockade activation cascade by immunosuppressive drugs prevents immune Here we studied whether TAC blocks peripheral human cells. After anti-CD3/CD28-activation cells healthy volunteers, (p65) phosphorylation was measured...

Tacrolimus has a large interindividual pharmacokinetic variability, and quantification of its effect is difficult. It substrate ABCB1, an efflux pump expressed more on CD8 T cells than CD4 cells. The ABCB1 3435C>T single-nucleotide polymorphism (SNP) been associated with differences in activity may influence drug efficacy. Here the this SNP biological tacrolimus was studied.Rhodamine (Rh123) used to study activity, or without addition inhibitor verapamil. Intracellular interleukin (IL) 2...

Monocytes and macrophages play key roles in many disease states, including cellular humoral rejection after solid organ transplantation (SOT). To suppress alloimmunity SOT, immunosuppressive drug therapy is necessary. However, little known about the effects of drugs tacrolimus mycophenolic acid (MPA) on monocyte activation function. Here, effect these immunosuppressants monocytes was investigated by measuring phosphorylation three intracellular signaling proteins which all have a major role...

The effect of immunosuppressive drugs on the generation T follicular helper (Tfh) cells, specialized in supporting B-cell differentiation, is largely unknown. We examined whether calcineurin inhibitor tacrolimus (TAC) and mammalian target rapamycin (mtor) sirolimus (SRL) inhibit Tfh cell affect subsequent functions. Isolated naive cells were polarized into Tfh-like presence TAC or SRL. To demonstrate their functionality, we co-cultured these with isolated B alloantigen studied activation...

Background The co-stimulatory inhibitor of the CD28-CD80/86-pathway, belatacept, allows calcineurin-inhibitor-free immunosuppression in kidney transplantation. However, aggressive T-cell mediated allogeneic responses have been observed belatacept-treated patients, which could be explained by effector-memory T-cells that lack membrane expression CD28, i.e. CD28-negative (CD28NULL) T-cells. CD28-positive (CD28POS) down regulate their surface CD28 after stimulation also pose a threat against...

B cell responses are dependent on specialized help provided by follicular T helper (Tfh). Tfh and cells both express the transcription factor B-cell lymphoma 6 protein (BCL6) targeting BCL6 may thus prevent humoral allo-immune after transplantation. In this study, effects of small molecule inhibitor 79-6 human differentiation proliferation were investigated. To end, naïve CD4 resting stimulated polyclonally in presence 79-6. This compound suppressed proliferation, differentiation, function...

Acute rejection is one of the major immunological determinants kidney graft function and survival. Early biomarkers to predict are lacking. Emerging evidence reveals a crucial role for monocyte/macrophage lineage cells in pathogenesis rejection. We hypothesized that higher pretransplant numbers proinflammatory CD16+ monocytes can The study cohort consisted 104 transplant recipients (58 with no 46 biopsy-proven rejection) 33 healthy persons. Posttransplant median follow-up time was 14.7 mo...

Humoral alloreactivity has been recognized as a common cause of kidney transplant dysfunction. B-cell activation, differentiation and antibody production are dependent on IL-21+ CXCR5+ follicular T-helper (Tfh) cells. Here, we studied whether belatacept, an inhibitor the co-stimulatory CD28-CD80/86-pathway, interrupts crosstalk between Tfh- B-cells more efficiently than calcineurin tacrolimus. The suppressive effects belatacept tacrolimus donor antigen-driven Tfh-B-cell interaction were...

In Brief Background Belatacept has been associated with an increased acute rejection rate after kidney transplantation. This case report sheds light on the possible immunological mechanisms underlying this phenomenon by analyzing in patient serum, peripheral blood mononuclear cells, rejected tissue, and graft infiltrating cells. Methods A 61-year-old woman treated belatacept, who received her first transplant from husband was admitted acute, vascular 56 days transplantation which...

Mesenchymal stem cells from pediatric patients (pMSCs) are an attractive cell source in regenerative medicine, due to their higher proliferation rates and better differentiation abilities compared adult MSCs (aMSCs). We have previously characterized the immunomodulatory of pMSCs on T under co-culture. It has also been reported that aMSCs can inhibit B maturation inflammatory conditions. In this study, we therefore aimed clarify effect towards microenvironment. Bone marrow derived were primed...

Rabbit anti-thymocyte globulins (rATG) induce CD4(+)CD25(+)forkhead box P3 (FoxP3(+)) regulatory T cells that control alloreactivity. In the present study, we investigated whether rATG convert into functional CD4(+)CD25(+)FoxP3(+)CD127(-/low) in presence of drugs may hamper their induction and function, i.e. calcineurin inhibitors. CD25(neg) were stimulated with or rabbit immunoglobulin G (rIgG) absence tacrolimus for 24 h. Flow cytometry was performed CD4, CD25, FoxP3 CD127 function CD25(+)...

BACKGROUND.: Rabbit antithymocyte globulins (rATGs) are known to convert CD4CD25FoxP3 T cells from healthy individuals cells. In this study, we investigated the effect of rATG on induction regulatory (Tregs) blood patients with end-stage renal disease who candidates for transplantation and rATG-induction therapy. The induced Tregs were analyzed compared naturally occurring CD4CD25FoxP3T METHODS.: CD25 pretransplant (n=7) controls (n=4) stimulated or control rabbit immunoglobulins 24 hr....

Expansion of Ag-specific naturally occurring regulatory T cells (nTregs) is required to obtain sufficient numbers for cellular immunotherapy. In this study, different allogeneic stimuli were studied their capacity generate functional alloantigen-specific nTregs. A highly enriched nTreg fraction (CD4(+)CD25(bright)CD127(-) cells) was expanded using HLA-mismatched immature, mature monocyte-derived dendritic (moDCs), or PBMCs. The moDC-expanded nTregs fully characterized by analysis the...

Proinflammatory, cytotoxic CD4+CD28null T cells can be substantially expanded in patients with end-stage renal disease. These have been associated the risk for rejection, but their alloreactive potential is unknown. were stimulated HLA-mismatched antigen presenting absence/presence of exogenous cytokines. Alloreactive was evaluated based on proliferation, degranulation, cytotoxicity, and cytokine production. Further, suppressive capacity assessed by measuring inhibition proliferating CD28+...

Background Therapeutic drug monitoring (TDM) of tacrolimus, based on blood concentrations, shows an imperfect correlation with the occurrence rejection. Here, we tested whether measuring NFATc1 amplification, a member calcineurin pathway, is suitable for TDM tacrolimus. Materials and methods amplification was monitored in T cells kidney transplant recipients who received either tacrolimus- (n = 11) or belatacept-based 10) therapy. Individual effects were studied vitro, after spiking samples...

Pharmacokinetic immunosuppressive drug monitoring poorly correlates with clinical outcomes after solid organ transplantation. A promising method for pharmacodynamic of tacrolimus (TAC) in T cell subsets transplant recipients might be the measurement (phosphorylated) p38MAPK, ERK1/2 and Akt (activated downstream receptor) by phospho-specific flow cytometry. Here, blood samples from n = 40 kidney (treated either TAC-based or belatacept (BELA)-based therapy) were monitored before throughout...

The novel immunosuppressant sotrastaurin is a selective inhibitor of protein kinase C isoforms that are critical in signalling pathways downstream the T cell receptor. Sotrastaurin inhibits nuclear factor (NF)-κB, which directly promotes transcription forkhead box 3 (FoxP3), key regulator for development and function regulatory cells (Tregs). Our center participated randomized trial comparing (n = 14) calcineurin Neoral 7) renal transplant recipients. We conducted ex vivo mixed lymphocyte...

Blockade of the CD80/86-CD28 pathway by belatacept after kidney transplantation is associated with an increased risk rejection compared standard, calcineurin inhibitor (CNI)-based therapy. CD28- T cells, which express CD57, are not susceptible to treatment. High numbers CD4+ CD57+ programmed death 1 (PD-1)- cells pretransplantation have been a higher chance rejection, although conflicting data reported. To investigate working mechanism behind this possible we studied expression co-inhibitory...

Monocytes significantly contribute to ischemia-reperfusion injury and allograft rejection after kidney transplantation. However, the knowledge about effects of immunosuppressive drugs on monocyte activation is limited. Conventional pharmacokinetic methods for drug monitoring are not cell type-specific. In this study, phosphorylation 3 signaling proteins was measured determine pharmacodynamic immunosuppression in transplant patients.Blood samples from 20 recipients were monitored before...