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Environmental seropositivity risk factors indicate natural ecosystems may have affected spread of the disease., The introduction of bluetongue virus serotype 8 into northern Europe at the end of summer 2006 initiated one of the most widespread epizootics of bluetongue infection ever to occur. In winter 2007–2008, a cross-sectional serologic study was conducted in France along a transect perpendicular to the epizootic wave. Cattle herd-level seroprevalence varied from 4% to 100%, and animal-level seroprevalence from

Several tumor antigens have been described as candidates for immunotherapy. Our study compared HLA-A2-restricted epitopes from 5 antigens commonly expressed by melanomas, i.e., Melan-A/MART-1 peptides (26–35 and 27–35), tyrosinase (368–376), gp-100 (280–288), MAGE-3 (271–279) and NA17-A (1–10), for their relative capacity to promote the development of cytotoxic and cytokine-producing specific CD8+ lymphocytes within melanoma-invaded lymph nodes. We used short-term cultured melanoma-invaded lymph node lymphocytes (MILLs) and tested responses developed by these cells to peptide-pulsed TAP-deficient T2 cells. We measured both the lytic response developed by MILLs and the fraction of these cells that secreted interferon-γ (IFN-γ), as deduced from intracellular cytokine labeling. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the expression of the 5 antigens within melanoma-invaded lymph nodes. Melan-A/MART-1, tyrosinase and gp-100 peptides were recognized by MILLs derived, respectively, from 8 of 20, 5 of 19 and 4 of 20 melanoma-invaded lymph nodes expressing these antigens. Most MILLs specific for Melan-A/MART-1 and tyrosinase exhibited both lysis and IFN-γ responses, whereas most of those specific for gp-100 developed only lysis. Weak lysis without IFN-γ secretion was developed against NA17-A and MAGE-3 peptides by MILLs from, respectively, 3 of 9 and 2 of 14 lymph nodes expressing these antigens. Our data show a prevalence of both cytotoxic and IFN-γ-secreting effector T cells specific for differentiation antigens within HLA-A2 melanoma-invaded lymph nodes, which makes these antigens attractive targets for specific immunotherapy. Int. J. Cancer 78:209–215, 1998.© 1998 Wiley-Liss, Inc.

Measurement of the immunoglobulin G concentration of dairy cow colostrum. A study was conducted to see if the concentration of colostral immunoglobulin G (IgG) in Holstein dairy cows differed according to the sampling method. Concentrations of IgG were measured by radial immune-diffusion (RID). Three sampling regimens were tested: composite sample, single sample and time of sampling. IgG was measured in: a composite sample (taken from the 4 quarters of the udder at first milking in 78 cows),in a single sample (taken from 1 quarter at first milking in 78 cows) or in several samples, 7 to 9 samples taken through out the duration of first milking in 9 cows, to test a potential ‘time of sampling’ effect. In conclusion, at the level of 50 g/L, which is considered to be “good quality” or acceptable colostrum, the concentration of IgG was unaffected by sampling time or sampletype. Based on the calves’ capacity to drink, observed in the present study, it is recommended that 2 to 4 L of colostrum be consumed within 2 hours of birth and that colostrum should be collected preferably from the posterior quarters. The quantity given is crucial due to the quality of dairy cow colostrum.; Une étude a été menée pour savoir si l’évaluation de la teneur en immunoglobulinesG (IgG) du colostrum chez la vache laitière Prim’Holstein différait, selon qu’elle se réalisait sur un échantillon de lait composite des quatre quartiers issus de la première traite (pratique courante), sur le lait issu d’un seul quartier, ou à plusieurs moments de la première traite. La teneur en IgG a été mesurée sur 78 vaches par immuno-diffusion radiale(IDR), à la fois sur un échantillon prélevé de la première traite des 4 quartiers(lait composite) et sur un échantillon des autres quartiers. De plus,9 vaches ont fait l’objet de 7 à 9 prélèvements en cours de première traite pour évaluer un effet potentiel « moment du prélèvement ». Il ressort qu’au seuil de 50 g/L pour considérer un colostrum de « bonne qualité »ou de qualité acceptable, il n’y avait aucune influence ni du moment de prélèvement, ni du type de prélèvement, sur la mesure de la teneur enIgG. Compte tenu de la capacité de buvée des veaux observée dans l’étude, il peut être recommandé de donner 2 à 4 litres dans les 2 premières heures de vie du veau, le colostrum pouvant être collecté préférentiellement des quartiers postérieurs. La quantité administrée est cruciale en raison de la qualité des colostrums de vaches laitières.

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Recently a commercial real-time reverse transcription polymerase chain reaction (RT-PCR) kit has been marketed for the detection of Bovine respiratory syncytial virus (BRSV). However, diagnostic interpretation of the results of this kit requires its comparison to commonly used methods. Therefore, the objective of this study was to evaluate the performance of this kit in comparison with the conventional direct fluorescent antibody test (FAT). Twenty BRSV strains and 14 heterologous bovine viruses were used to check the kit's sensitivity and specificity. The efficiency and detection limit of the kit were determined by testing dilution series of a BRSV strain. The comparison between real-time RT-PCR kit and FAT was performed with 94 clinical samples from calves with clinical signs of respiratory disease including lung tissues (n = 55), transtracheal aspiration samples (n = 20), and nasal swab samples (n = 19). All of the BRSV strains tested were detected by real-time RT-PCR. No cross-reaction was shown with the 14 heterologous bovine viruses. The real-time RT-PCR was 99.3% efficient with a detection limit of 0.1 TCID50 (50% tissue culture infective dose). The results of real-time RT-PCR and FAT were concordant for 65 of the 94 clinical samples tested. The remaining 29 clinical samples were positive by real-time RT-PCR and negative by FAT, demonstrating the higher sensitivity of real-time RT-PCR. In conclusion, the kit evaluated in this study was sensitive, specific, and had a low threshold of detection. Furthermore, the use of this kit instead of FAT allows an improvement of the sensitivity for the detection of BRSV in clinical samples.

T cell activation is known to depend not only on efficient antigen recognition and subsequent signaling through TCR, but also on interactions involving multiple adhesion and accessory molecules such as CD28/B7, LFA-1/ICAM-1 and LFA-3/CD2. The present study dissects the role of LFA-3/CD2 interactions in the activation of melanoma-specific CD8+ T cell clones. To this end we analyzed the influence of LFA-3 density on melanoma cells on lysis and cytokine production (TNF, IL-2, IFN-gamma) by T cells following activation by various amounts of antigenic peptides. Our results indicate that increasing LFA-3 density on melanoma cells variably affects their lysis susceptibility, but systematically and considerably enhances cytokine production by melanoma-specific cytotoxic T lymphocyte (CTL) clones. At any stimulatory antigen density, LFA-3 increased the fraction of responding cells and/or cytokine amounts produced by individual cells, without affecting TCR down-regulation. These results show that CD2 engagement increases cytokine gene activation essentially by providing to T cells a TCR-independent co-activation signal. From a practical point of view, our data demonstrate that the level of LFA-3 expressed on tumors critically affects cytokine production by specific CTL and thus the efficiency of specific immune reactions mediated by these cells.

International audience; Subsets of T and natural killer (NK) lymphocytes express the CD94-NKG2A heterodimer, a receptor for major histocompatibility complex class I molecules. We show here that engagement of the CD94-NKG2A heterodimer inhibits both antigen-driven tumor necrosis factor (TNF) release and cytotoxicity on melanoma-specific human T cell clones. Similarly, CD16-mediated NK cell cytotoxicity is extinguished by cross-linking of the CD94-NKG2A heterodimer. Combining in vivo and in vitro analysis, we report that both I/VxYxxL immunoreceptor tyrosine-based inhibition motifs (ITIM) present in the NKG2A intracytoplasmic domain associate upon tyrosine phosphorylation with the protein tyrosine phosphatases SHP-1 and SHP-2, but not with the polyinositol phosphatase SHIP Determination of the dissociation constant, using surface plasmon resonance analysis, indicates that NKG2A phospho-ITIM interact directly with the SH2 domains of SHP-1 and SHP-2 with a high affinity. Engagement of the CD94-NKG2A heterodimer therefore appears as a protein-tyrosine phosphatase-based strategy that negatively regulates both antigen-induced T cell response and antibody-induced NK cell cytotoxicity. Our results suggest that this inhibitory pathway sets the threshold of T and NK cell activation.

International audience; CD4+ T cell clones derived from lymphocytes infiltrating four human melanomas specifically recognized melanoma-derived tumor epitopes as shown by secretion of tumor necrosis factor (TNF) in vitro upon interaction with autologous melanoma cells, whereas they did not recognize HLA class II-expressing autologous lymphoblasts or HLA class II mismatched allogeneic melanoma cells. Specificity was further established by demonstrating that TNF responses to tumor cells were inhibited by HLA-DR or HLA-DQ monoclonal antibodies. Most of these clones cross-reacted with allogeneic melanoma cells expressing a potentially restricting HLA allele or a structurally similar one. These data show that shared epitopes of human melanoma cells presented on HLA class II molecules are frequently recognized by autologous CD4+ T lymphocytes.

International audience; Human melanomas are infiltrated by tumor-reactive T lymphocytes. However, the ability of these cells to elicit a specific anti-tumor response in vivo remains to be established. Because lymphokine production is critical for T cell functions, we have analyzed the capacity of melanoma-specific tumor-infiltrating lymphocyte (TIL) clones to produce major lymphokines: interleukin-2 (IL-2), interferon-gamma (IFN-gamma) and interleukin-4 (IL-4), as well as tumor necrosis factor (TNF), in response to direct antigen presentation by autologous and allogeneic tumor cells. We report here that, upon stimulation by autologous melanoma cells, all TIL clones secreted TNF but only a few of them produced significant amounts of IL-2, IL-4 or IFN-gamma. Nonetheless, all these clones consistently produced two or three of these last lymphokines upon stimulation with phorbol myristate acetate and calcium ionophore, as well as IL-2 upon CD3 stimulation, showing the existence of three lymphokine profiles among them: Th1, Th0 and a profile characterized by IL-2 and IL-4, but not IFN-gamma secretion. Stimulation of TIL clones by allogeneic melanoma lines sharing the appropriate HLA-peptide complexes revealed that defective IL-2 production seemed to be a constant feature for some clones, while it was, for other clones, dependent on the antigen-presenting tumor cells. For this last type of clone, we further showed that defective IL-2 induction resulted from an LFA-3 defect of some melanoma cells or from distinct yet undefined defects of other melanoma lines. Our data suggest that defective lymphokine secretion may be an essential component of the in vivo failure of melanoma-reactive TIL to control tumor development. Interestingly both CD4+ and CD8+ TIL clones from one patient were fully activated by the autologous melanoma cells in vitro, supporting a potential role of such TIL in spontaneous or induced tumor rejection.