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Optimization of in vitro expansion of macaque CD4 + T cells using anti‐CD3 and co‐stimulation for autotransfusion therapy
Author(s) -
Onlamoon Nattawat,
Hudson Krystal,
Bryan Patsy,
Mayne Ann E.,
Bonyhadi Mark,
Berenson Ron,
Sundstrom Bruce J.,
Bostik Pavel,
Ansari Aftab A.,
Villinger François
Publication year - 2006
Publication title -
journal of medical primatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.31
H-Index - 42
eISSN - 1600-0684
pISSN - 0047-2565
DOI - 10.1111/j.1600-0684.2006.00182.x
Subject(s) - cd154 , cd28 , t cell , biology , in vitro , cd3 , stimulation , adoptive cell transfer , microbiology and biotechnology , immunology , cd40 , antigen , immune system , cytotoxic t cell , cd8 , endocrinology , biochemistry
Background Our laboratory has previously shown that adoptive transfer of in vitro ‐expanded autologous purified polyclonal CD4 + T cells using anti‐CD3/CD28‐coated beads induced antiviral responses capable of controlling SIV replication in vivo . Methods As CD4 + T cells comprise several phenotypic and functional lineages, studies were carried out to optimize the in vitro culture conditions for maximal CD4 + T‐cell expansion, survival and delineate the phenotype of these expanded CD4 + T cells to be linked to maximal clinical benefit. Results and Conclusions The results showed that whereas anti‐monkey CD3 γ / ɛ was able to induce T‐cell proliferation and expansion in combination with antibodies against multiple co‐stimulatory molecules, monkey CD3 ɛ cross reacting antibodies failed to induce proliferation of macaque CD4 + T cells. Among co‐stimulatory signals, anti‐CD28 stimulation was consistently superior to anti‐4‐1BB, CD27 or ICOS while the use of anti‐CD154 failed to deliver a detectable proliferation signal. Increasing the relative anti‐CD28 co‐stimulatory signal relative to anti‐CD3 provided a modest enhancement of expansion. Additional strategies for optimization included attempts to neutralize free radicals, enhancement of glucose uptake by T cells or addition of T‐cell stimulatory cytokines. However, none of these strategies provided any detectable proliferative advantage. Addition of 10 autologous irradiated feeder cells/expanding T cell provided some enhancement of expansion; however, given the high numbers of T cell needed, this approach was deemed impractical and costly, and lower ratios of feeder to expanding T cells failed to provide such benefit. The most critical parameter for efficient expansion of purified CD4 + T cells from multiple monkeys was the optimization of space and culture conditions at culture inception. Finally, anti‐CD3/28‐expanded CD4 + T cells uniformly exhibited a central memory phenotype, absence of CCR5 expression, marked CXCR4 expression in vitro , low levels of caspase 3 but also of Bcl‐2 expression.