Heron I, Schreier M, Cahill R, Poskitt D, Trnka Z
J Immunol Methods. 1978;24(3-4):311-20. doi: 10.1016/0022-1759(78)90134-5.
The in vitro induction of specific primary and secondary immune responses in sheep lymph node cell suspensions is described and suitable culture conditions determined. The induction of primary immune responses required supplementation of the culture medium with antigen-absorbed homologous serum or lymph, whereas the requirements for the induction of a secondary response were less stringent. The addition of 2-mercaptoethanol to the medium was required. The amounts of heterologous erythrocytes used for immunization were critical and optimal responses were obtained when 50 microleters of a 1% suspension were added to 1 ml cultures. Lymphocyte densities of about 5 X 10(6)/ml were found optimal in primary immune responses in vitro. Less than 2 X 10(6) cells/ml rarely gave rise to plaque-forming cell (PFC) generation, whereas densities of 10 x 10(6) and above reduced the number of PFC obtained per number of cultured cells. Lymphocytes obtained from the efferent lymph draining lymph nodes previously immunized with heterologous erythrocytes were found to generate PFC in vitro when specific antigen was added to the cultures, but attempts to generate PFC in vitro with cells from efferent lymph draining non-immunized nodes failed.
本文描述了绵羊淋巴结细胞悬液中特异性初次和二次免疫反应的体外诱导过程，并确定了合适的培养条件。初次免疫反应的诱导需要在培养基中添加抗原吸收的同源血清或淋巴液，而二次反应诱导的要求则不那么严格。培养基中需要添加2-巯基乙醇。用于免疫的异源红细胞数量至关重要，当向1ml培养物中加入50微升1%的悬液时可获得最佳反应。体外初次免疫反应中，淋巴细胞密度约为5×10⁶/ml时最为适宜。每毫升少于2×10⁶个细胞很少能产生空斑形成细胞（PFC），而密度为10×10⁶及以上时，每培养细胞获得的PFC数量会减少。当向培养物中添加特异性抗原时，发现从先前用异源红细胞免疫的淋巴结引流的输出淋巴中获得的淋巴细胞可在体外产生PFC，但尝试用未免疫淋巴结引流的输出淋巴中的细胞在体外产生PFC却失败了。
