Human T lymphocyte activation kinetics for identifying and targeting alloreactive T cells

摘要: Selective depletion of alloreactive T cells from a stem cell graft has the potential reducing versus host disease (GvHD) while preserving malignancy and third party responses. For this purpose several techniques generate deplete cells, which are donor-derived activated by recipient tissue. The kinetics activation in donor-recipient co-culture systems is critical optimizing timing cells. We present our preclinical system. Peripheral blood mononuclear (PBMCs) were derived pairs unrelated healthy human volunteers. 2500 cGy irradiated (stimulators) co-cultured with PBMCs (responders) 1:1 ratio concentration 5 x 10 6 /ml serum free medium. Stimulator labeled PKH67 co-cultures were, analyzed for CD3, CD4, CD25, expression flow cytometry on days 0 through 7, using Topro-3 to exclude dead also added low dose Interleukin-2 (IL2) augmenting generation see if made difference. Our results show that CD8 (CD3+, CD4−, CD25+) increased ≤1 % day 0, 6.5 percent 5; addition IL2 amplified nearly 10% 5. CD4 + CD4+, CD25+dim or total) did not appreciably increase over time ranged between 2 5%; have any effect. regulatory CD25+bright) at baseline 5% these unaffected IL2. proportion non-activated lymphocytes, decreased progression. lymphocyte activation, defined CD25 expression, progressively increases first week co-culture. This important observation will help establishing allograft manipulation, selective clinical hematopoietic transplantation.