Micro-system for the primary immunization of baboon (Papio cynocephalus) peripheral blood mononuclear cells with sheep erythrocytes in vitro.

A micro-culture system for the stimulation of baboon peripheral blood mononuclear cells (PBMC) with sheep erythrocytes (SRBC) in vitro was established. PBMC cultures in 96-well microtiter plates were maintained in 300 microliter of culture medium containing a murine mixed lymphocyte reaction (MLR) supernatant and SRBC. Cultures of 7.5 x 10(5) PBMC and 6 x 10(6) SRBC resulted in the highest anti-SRBC plaque-forming cell (PFC) response. It was also determined that the presence of 50 microliter of murine MLR supernatant was required for optimal PFC generation and that the cultures required supplementation with nutrient cocktail 3 days post-immunization. Although PFC were detectable on days 4-8, the maximum expression of PFC occurred on day 7.