IL-21 promotes survival and maintains a naive phenotype in human CD4+ T lymphocytes

IL-21 is a key T-cell growth factor (TCGF) involved in innate and adaptive immune response. It contributes to the proliferation of naive, but not memory T lymphocytes. However, the full spectrum of IL-21 activity on T cells remains unclear. Here, we demonstrate that IL-21 primarily maintains the expression of specific naive cell surface markers such as CD45RA, CD27, CD62L and CCR7 on human CD4+ T lymphocytes and that the expression of CCR7 induces cell migration by means of CCL21 chemoattraction. These effects contrast with those of IL-2 which induced the marked proliferation of CD4+ T lymphocytes, leading to an activated-memory phenotype. Nevertheless, IL-21 maintained cell cycle activation and expression of proliferation markers, including proliferating cell nuclear antigen and Ki-67, and triggered T-cell proliferation via TCR and co-stimulation pathways. Unlike IL-2, IL-21 decreased the expression of the anti-apoptotic Bcl-2 protein, which correlated with the absence of activation of the phosphatidylinositol 3′-kinase/Akt signaling pathway. Thus, IL-21 is a TCGF whose function is the preservation of a pool of CD4+ T lymphocytes in a naive phenotype, with a low proliferation rate but with the persistence of cell cycling proteins and cell surface expression of CCR7. These findings strongly suggest that IL-21 plays a part in innate and adaptive immune response owing to homeostasis of T cells and their homing to secondary lymphoid organ

The proinflammatory cytokines IL-2, IL-15 and IL-21 modulate the repertoire of mature human natural killer cell receptors

Natural killer (NK) cells play a crucial role in the immune response to micro-organisms and tumours. Recent evidence suggests that NK cells also regulate the adaptive T-cell response and that it might be possible to exploit this ability to eliminate autoreactive T cells in autoimmune disease and alloreactive T cells in transplantation. Mature NK cells consist of a highly diverse population of cells that expresses different receptors to facilitate recognition of diseased cells and possibly pathogens themselves. Ex vivo culture of NK cells with cytokines such as IL-2 and IL-15 is an approach that permits significant expansion of the NK cell subpopulations, which are likely to have potent antitumour, antiviral, or immunomodulatory effects in autoimmunity. Our data indicate that the addition of IL-21 has a synergistic effect by increasing the numbers of NK cells on a large scale. IL-2 and IL-15 may induce the expression of killer cell immunoglobulin-like receptors (KIRs) in KIR-negative populations, the c-lectin receptor NKG2D and the natural cytotoxic receptor NKp44. The addition of IL-21 to IL-15 or IL-2 can modify the pattern of the KIR receptors and inhibit NKp44 expression by reducing the expression of the adaptor DAP-12. IL-21 also preserved the production of interferon-γ and enhanced the cytotoxic properties of NK cells. Our findings indicate that the proinflammatory cytokines IL-2, IL-15 and IL-21 can modify the peripheral repertoire of NK cells. These properties may be used to endow subpopulations of NK cells with specific phenotypes, which may be used in ex vivo cellular immunotherapy strategies

Systematic mapping of regions of human cardiac troponin I involved in binding to cardiac troponin C: N- and C-terminal low affinity contributing regions

AbstractThe Spot method of multiple peptide synthesis was used to map in a systematic manner regions of the human cardiac troponin I sequence (hcTnI) involved in interactions with its physiological partner, troponin C (cTnC). Ninety-six 20-mer peptides describing the entire hcTnI sequence were chemically assembled; their reactivity with [125I]cTnC, in the presence of 3 mM Ca2+, enabled the assignment of six sites of interaction (residues 19–32, 45–54, 129–138, 145–164, 161–178 and 191–210). For several sites, a good correlation with literature data was obtained, thus validating this methodological approach. Synthetic peptides, each containing in their sequence an interaction site, were prepared. As assessed by BIACORE, all of them exhibited an affinity for cTnC in the range of 10−6–10−7 M, except for hcTnI [39–58] which showed a nanomolar affinity. This peptide was also able to block the interaction between hcTnI and cTnC. We therefore postulate that despite the existence of multiple cTnC interaction sites on the hcTnI molecule, only that region of hcTnI allows a stabilization of the complex. Residues 19–32 from the N-terminal cardio-specific extension of hcTnI were also found to be involved in interaction with cTnC; residues 19–32 may correspond to the minimal sequence of the extension which could switch between the N- and C-terminal TnC domains, depending on its phosphorylation state. Finally, two Ca2+-dependent cTnC binding domains within the C-terminal part of hcTnI (residues 164–178 and 191–210) were also mapped. The latter site may be linked with the cardiac dysfunction observed in stunned myocardium

PEPOP: Computational design of immunogenic peptides

© 2008 Moreau et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution Licens

The impact of pre-transplant donor specific antibodies on the outcome of kidney transplantation - Data from the Swiss transplant cohort study

Background Pre-transplant donor specific antibodies (DSA), directed at non-self human leukocyte antigen (HLA) protein variants present in the donor organ, have been associated with worse outcomes in kidney transplantation. The impact of the mean fluorescence intensity (MFI) and the target HLA antigen of the detected DSA has, however, not been conclusively studied in a large cohort with a complete virtual cross-match (vXM). Methods We investigated the effect of pre-transplant DSA on the risk of antibody-mediated rejection (ABMR), graft loss, and the rate of eGFR decline in 411 DSA positive transplants and 1804 DSA negative controls. Results Pre-transplant DSA were associated with a significantly increased risk of ABMR, graft loss, and accelerated eGFR decline. DSA directed at Class I and Class II HLA antigens were strongly associated with increased risk of ABMR, but only DSA directed at Class II associated with graft loss. DSA MFI markedly affected outcome, and Class II DSA were associated with ABMR already at 500-1000 MFI, whereas Class I DSA did not affect outcome at similar low MFI values. Furthermore, isolated DSA against HLA-DP carried comparable risks for ABMR, accelerated eGFR decline, and graft loss as DSA against HLA-DR. Conclusion Our results have important implications for the construction and optimization of vXM algorithms used within organ allocation systems. Our data suggest that both the HLA antigen target of the detected DSA as well as the cumulative MFI should be considered and that different MFI cut-offs could be considered for Class I and Class II directed DSA

The impact of pre-transplant donor specific antibodies on the outcome of kidney transplantation - Data from the Swiss transplant cohort study.

Background Pre-transplant donor specific antibodies (DSA), directed at non-self human leukocyte antigen (HLA) protein variants present in the donor organ, have been associated with worse outcomes in kidney transplantation. The impact of the mean fluorescence intensity (MFI) and the target HLA antigen of the detected DSA has, however, not been conclusively studied in a large cohort with a complete virtual cross-match (vXM). Methods We investigated the effect of pre-transplant DSA on the risk of antibody-mediated rejection (ABMR), graft loss, and the rate of eGFR decline in 411 DSA positive transplants and 1804 DSA negative controls. Results Pre-transplant DSA were associated with a significantly increased risk of ABMR, graft loss, and accelerated eGFR decline. DSA directed at Class I and Class II HLA antigens were strongly associated with increased risk of ABMR, but only DSA directed at Class II associated with graft loss. DSA MFI markedly affected outcome, and Class II DSA were associated with ABMR already at 500-1000 MFI, whereas Class I DSA did not affect outcome at similar low MFI values. Furthermore, isolated DSA against HLA-DP carried comparable risks for ABMR, accelerated eGFR decline, and graft loss as DSA against HLA-DR. Conclusion Our results have important implications for the construction and optimization of vXM algorithms used within organ allocation systems. Our data suggest that both the HLA antigen target of the detected DSA as well as the cumulative MFI should be considered and that different MFI cut-offs could be considered for Class I and Class II directed DSA

How Morphological Constraints Affect Axonal Polarity in Mouse Neurons

Neuronal differentiation is under the tight control of both biochemical and physical information arising from neighboring cells and micro-environment. Here we wished to assay how external geometrical constraints applied to the cell body and/or the neurites of hippocampal neurons may modulate axonal polarization in vitro. Through the use of a panel of non-specific poly-L-lysine micropatterns, we manipulated the neuronal shape. By applying geometrical constraints on the cell body we provided evidence that centrosome location was not predictive of axonal polarization but rather follows axonal fate. When the geometrical constraints were applied to the neurites trajectories we demonstrated that axonal specification was inhibited by curved lines. Altogether these results indicated that intrinsic mechanical tensions occur during neuritic growth and that maximal tension was developed by the axon and expressed on straight trajectories. The strong inhibitory effect of curved lines on axon specification was further demonstrated by their ability to prevent formation of multiple axons normally induced by cytochalasin or taxol treatments. Finally we provided evidence that microtubules were involved in the tension-mediated axonal polarization, acting as curvature sensors during neuronal differentiation. Thus, biomechanics coupled to physical constraints might be the first level of regulation during neuronal development, primary to biochemical and guidance regulations

PTX3 Polymorphisms and Invasive Mold Infections After Solid Organ Transplant

Donor PTX3 polymorphisms were shown to influence the risk of invasive aspergillosis among hematopoietic stem cell transplant recipients. Here, we show that PTX3 polymorphisms are independent risk factors for invasive mold infections among 1101 solid organ transplant recipients, thereby strengthening their role in mold infection pathogenesis and patients' risk stratificatio

Neutralisation des anticorps inhibiteurs survenant chez les hémophiles A traités par le facteur VIII

Le facteur VIII est un des éléments clés de la cascade de la coagulation sanguine. De nombreux individus sont atteints d'un défaut quantitatif ou qualificatif de l'expression de cette molécule : ce sont les hémophiles A, qui doivent recevoir pour survivre du facteur VIII par transfusion. Dans la majorité des cas ( en dehors des problèmes de contamination virale), ce traitement est sans complication. Toutefois, il s'avère que certains hémophiles développent une réponse immunitaire contre la molécule injectée. L'apparition de ces anticorps inhibe l'activité procoagulante de FVIII et altère l'efficacité du traitement. Les thérapies actuelles pour contourner cette complication sont actuellement d'un coût très élevé et leur efficacité se limite aux patients présentant un faible titre inhibiteurs. Une approche thérapeutique raisonnable au problème posé par ces anticorps anti-FVIII est de parvenir à en neutraliser les effets inhibiteurs en bloquant le site de liaison de ces anticorps. Pour ce faire, nous avons retenu deux stratégies : la première repose sur une régulation de type idiotypique, qui consisterait à manipuler la réponse immunitaire du patient pour lui faire exprimer une réponse anti-idiotypique neutralisant les effets non désirables des anti-FVIII, permettant ainsi de restaurer l'activité procoagulante du FVIII. Dans une étude préliminaire, nous avons caractérisé plusieurs anticorps monoclonaux inhibiteurs murins, considérés comme de bons représentants des anticorps inhibiteurs humains. Cela comprend la détermination de leur spécificité d'épitopes, le séquençage de leurs parties variables ( article V ), ainsi que l'élucidation de leur mécanisme d'inactivation ( article I ). Ces anticorps anti-FVIII ont permis de préparer des anticorps polyclonaux anti-idiotypiques chez le lapin. Ceux-ci présentent des propriétés neutralisantes de l'activité inhibitrice de leur anticorps respectif à la fois in vitro et dans des tests fonctionnels. Ces anticorps anti-idiotypiques sont surtout capables de lier spécifiquement des anticorps anti-FVIII contenus chez des patients hémophiles, laissant entrevoir un potentiel thérapeutique à cette stratégie ( article II). La seconde approche évaluée repose sur l'hypothèse que le blocage des anticorps pathogènes par des substituts antigéniques du FVIII (peptides) pourrait améliorer la condition des hémophiles avec inhibiteurs. Pour sélectionner de tels peptides, des banques de phages peptidiques ont été utilisées. Pour la validation du concept, deux anticorps inhibiteurs dirigés contre le domaine C2 du FVIII, l'un murin l'anticorps ESH8 ( article III ), l'autre humain, l'anticorps B02C11 ( article IV ), dont les mécanismes d'inactivation sont suspectés être représentatifs de ceux des patients hémophiles avec inhibiteurs, ont été retenus comme cible. Dans les deux cas, des peptides capables sous forme soluble de bloquer l'activité inhibitrice d'ESH8 comme B02C11 in vitro et dans des tests fonctionnels ont été identifiés. Ces mêmes peptides s'avèrent aptes à neutraliser l'activité inhibitrice des deux anticorps in vivo dans un modèle murin représentatif de la pathologie de l'hémophilie A, ouvrant là aussi, de nouvelles perspectives thérapeutiques. Ces derniers résultats ont donné lieu à une demande de dépôt de brevet.MONTPELLIER-BU Pharmacie (341722105) / SudocPARIS-BIUP (751062107) / SudocSudocFranceF