Persistence of adoptively transferred allogeneic lymphocytes in recipient animals.

<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

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

<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

Experimental groupings and MHC genotype of cynomolgus macaques.

<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.

<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.

<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

Plasma α-defensin, IL-10 and IFN-γ levels with respect to the pre-infection values in naïve (n = 11) and adjuvant treated monkeys (n = 10) inoculated with SHIV_SF162P4cy.

<p>No significant differences were observed between naive and adjuvant treated monkeys during the observation period. Mean values with standard deviations are connected by dotted lines.</p

Effects of MHC class I<i>A</i> and I<i>B</i> haplotypes on IgG anti-Env antibodies and neutralizing Ab following infection with SHIV_SF162P4cy.

<p>Mean values with standard error are depicted for all animals positive or negative for the indicated haplotype: M4 (n = 6) non-M4 (n = 15) class I<i>A</i>; M4 (n = 7), non-M4 (n = 14) class I<i>B</i>.</p

Dynamics of viral infection in 21 cynomolgus monkeys inoculated with SHIV_SF162P4cy during acute (2–4 weeks p.i.), post-acute (8–16 weeks p.i.) of infection.

<p>Data represent mean values with standard error of log plasma RNA load, log proviral DNA, IgG anti-Env Ab, and nAb from 0 to 16 weeks p.i. (*) week 4: anti-Env bAb titers correlated positively with viral load (p = 0.0002); (**) week 16: nAb titers correlated positively with anti-Env bAb titers (p = 0.0041); (***) post-acute phase: proviral DNA levels correlated positively with anti-Env bAb (p = 0.0225) and negatively with nAb titers (p = 0.0083).</p

Effects of MHC class I<i>A</i> and II haplotypes on IL-10 and α-defensin production during SHIV_SF162P4cy infection.

<p>Mean values with standard error are depicted for all animals positive or negative for the indicated haplotype: M3 (n = 9) non-M3 (n = 12) class I<i>A</i>; M3 (n = 13) non-M3 (n = 8) class II.</p

Regression analyses between immunological and virological parameters in infected monkeys during the acute phase of SHIV_SF162P4cy infection.

<p>A significant relationship was detected between: A) IL-10 production and plasma viral load (p = 0.0023); B) α-defensins production and viral RNA (p = 0.0286); C) IFNγ and IL-10 production (p<0.0001). D) AUC: significant relationship between viral load and α-defensins (p = 0.0002).</p