15 research outputs found

    Allogeneic Lymphocyte Transfer in MHC-Identical Siblings and MHC-Identical Unrelated Mauritian Cynomolgus Macaques

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    <div><p>The detailed study of immune effector mechanisms in primate models of infectious disease has been limited by the inability to adoptively transfer lymphocytes from vaccinated animals into naïve immunocompetent recipients. Recent advances in our understanding of the Major Histocompatibility Complex diversity of Mauritian cynomolgus macaques enabled the establishment of a breeding program to generate Major Histocompatibility Complex (MHC)-identical animals. The current study utilised this resource to achieve an improved model of adoptive transfer of lymphocytes in macaques. The effect of route of transfusion on persistence kinetics of adoptively transferred lymphocytes was evaluated in an autologous transfer system. Results indicated that peripheral persistence kinetics were comparable following infusion by different routes, and that cells were detectable at equivalent levels in lymphoid tissues six weeks post-infusion. In a pilot-scale experiment, the persistence of adoptively transferred lymphocytes was compared in MHC-identical siblings and MHC-identical unrelated recipients. Lymphocytes transferred intra-peritoneally were detectable in the periphery within one hour of transfer and circulated at detectable levels in the periphery and lymph nodes for 10 days. Donor lymphocytes were detectable at higher levels in MHC-identical siblings compared with unrelated animals, however the total time of persistence did not differ. These results demonstrate a further refinement of the lymphocyte adoptive transfer system in Mauritian cynomolgus macaques and provide a foundation for hitherto impractical experiments to investigate mechanisms of cellular immunity in primate models of infectious disease.</p></div

    Persistence of adoptively transferred allogeneic lymphocytes in recipient animals.

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    <p>Solid lines represent peripheral blood samples; individual symbols represent lymph node biopsy samples. Dashed line indicates threshold for detection of CFDA-SE+ cells, calculated based on the mean of all negative samples plus two standard deviations. Animals are colour-coded by experimental group: <i>black</i> MHC-identical siblings; <i>red</i> MHC-identical unrelated; <i>blue</i> MHC-mismatch; <i>neg</i> negative control untreated animals.</p

    Experimental groupings and MHC genotype of cynomolgus macaques.

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    <p><i>rec</i>, recombinant MHC haplotype; a, reported ages and weights were on the day of allogeneic lymphocyte transfer.</p

    Detection of adoptively transferred allogeneic lymphocytes in lymphoid tissues of recipient animals at days 22, 35 and 36 post-transfer.

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    <p>Dashed line indicates threshold for detection of CFDA-SE+ cells. Animals are colour-coded by experimental group: <i>black</i> MHC-identical siblings; <i>red</i> MHC-identical unrelated; <i>blue</i> MHC-mismatched.</p

    Persistence of adoptively transferred autologous lymphocytes.

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    <p><b><i>A</i></b> Percentage of CFDA-SE+ lymphocytes in peripheral blood. Individual symbols represent lymph node biopsy samples; <b><i>B</i></b> Median fluorescence intensity of CFDA-SE+ cells; <b><i>C</i></b> Percentage of lymphocyte sub-populations positive for CFDA-SE; <b><i>D</i></b> Percentage of CD3+ and CD20+ cells in lymphocyte fraction.</p

    Macaque T cell cytokine responses to empty MV.

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    <p>Total cytokine (sum of TNFα+, IL-2+ and IFNγ+ cells) response to MV by CD4+ and CD8+ T cells from peripheral blood (PBMCs), peripheral lymph nodes (PLN), mesenteric lymph nodes (MLN) and spleen (SPL). Time in days is shown on x-axis, all responses from PBMC unless prefixed by a tissue. Group A, macaques F51–F54, were immunised with 10<sup>5</sup> TCID<sub>50</sub> MV1-F4 on days 0 and 28 whilst group B, macaques F55–F58, were immunised on day 0 only. Panels A and B shows cytokine responses gated on CD4+ T cells. Panels C and D show cytokine responses gated on CD8+ T cell. Dashed line represents the mean plus 3 standard deviations of day 0 responses, above which points were deemed significant.</p

    Macaque T cell cytokine responses to HIV-1 F4 peptides.

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    <p>Total cytokine (sum of TNFα+, IL-2+ and IFNγ+ cells) response to F4 peptides by CD4+ and CD8+ T cells from peripheral blood (PBMCs), peripheral lymph nodes (PLN), mesenteric lymph nodes (MLN) and spleen (SPL). Time in days is shown on x-axis, all responses from PBMC unless prefixed by a tissue. Group A, macaques F51–F54, were immunised with 10<sup>5</sup> TCID<sub>50</sub> MV1-F4 on days 0 and 28 whilst group B, macaques F55–F58, were immunised on day 0 only. Panels A and B shows cytokine responses gated on CD4+ T cells. Panels C and D show cytokine responses gated on CD8+ T cell. Dashed line represents the mean plus 3 standard deviations of day 0 responses, above which points were deemed significant.</p

    Macaque MHC haplotype analysis.

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    <p>MHC haplotypes of MV1-F4 vaccinees, F51–F58, were determined by microsatellite analysis and haplotypes with recombination were resolved by allele-specific PCR. Intact haplotypes, M1–M7, have been previously identified in Mauritian cynomolgus macaques <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050397#pone.0050397-Wiseman1" target="_blank">[62]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050397#pone.0050397-Mee2" target="_blank">[63]</a>.</p
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