Abstract
In vitro sensitization of human peripheral blood mononuclear cells is potentially useful in evaluation of immune responses, for hybridoma production, and for imnunogenetic and vaccine research. Secondary (2 °) in vitro lymphoproliferative responses are obtained after in vivo sensitization. However, primary (1°) in vitro immunization with soluble antigens has been more difficult. We have now induced 1° and 2° responses in vitro for purified antigens, complex soluble antigens, a lipid antigen presented in liposomes, and bacteria. PBMC were incubated at 106 /ml in flasks for 9-11 d with a 5 log concentration range of BSA, KLH, PPD, coccidioidin, liposomes with M. leprae phenolic glycolipid 1 (PG-1-L), or irradiated M. leprae. After washing, cells in microtiter plates were restimulated with antigen or fed without antigen. After 48 h cells were pulsed with H-thymidine. Results are shown as ratio CPM with restimulation:CPM without restimulation. Ratios ≥ 1.9 reflect lymphoproliferative responses; ratios ≤ 0.3 indicate suppression of responses (both p < .05). Sensitization was designated as 1° if prior reactivity by skin test and/or lymphocyte transformation was absent, or 2° if present. Responses were 1° for BSA (maximum ratio = 2.8), KLH (2.7), PPD (10.6), coccidioidin (2.5), and PG-1-L (6.3); 2° responses occurred for KLH, PPD, and M. leprao. 1° suppression was seen for BSA, coccidiodin, and PPD. The technique may be useful in inducing immune responses without exposing the subject.
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Schauf, V., Bengtson, K. & Holobauqh, P. HUMAN IMMUNIZATION IN VITRO. Pediatr Res 21 (Suppl 4), 317 (1987). https://doi.org/10.1203/00006450-198704010-00901
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DOI: https://doi.org/10.1203/00006450-198704010-00901