Booth R J
J Immunol Methods. 1979;26(3):253-63. doi: 10.1016/0022-1759(79)90250-3.
Small numbers of Ficoll-Hypaque purified human tonsillar lymphocytes were stimulated with PWM to produce SRBC-specific PFC in a microculture system. The magnitude of the response varied among different tonsils but was typically between 200 and 1000 PFC/10(6) cells cultured. Little or no response was observed in the absence of PWM. SRBC failed to stimulate a SRBC-specific response and the presence of this antigen in PWM-stimulated cultures depressed the response. The time of the maximum response was inversely related to the number of cells cultured. In addition, the duration of the response was limited by rapid depletion of critical medium requirements and/or build up of inhibitory factors especially when the cell concentration exceeded 5 x 10(5) cells/culture. This effect could be partially overcome by daily feeding of cultures with fresh medium. Fractionation studies indicated a requirement for both T and B cell populations. Constant efficiency of PFC production with respect to cell number could be achieved by the addition of inactivated autologous 'filler' cells. The significance of these results and applicability of the microculture system to a detailed analysis of human antibody responses will be discussed.
在微培养系统中，用美洲商陆有丝分裂原（PWM）刺激少量经聚蔗糖-泛影葡胺纯化的人扁桃体淋巴细胞，以产生抗绵羊红细胞（SRBC）特异性的空斑形成细胞（PFC）。不同扁桃体的反应强度各不相同，但通常在每10⁶个培养细胞产生200至1000个PFC之间。在没有PWM的情况下，几乎观察不到反应。SRBC未能刺激出SRBC特异性反应，且在PWM刺激的培养物中存在该抗原会抑制反应。最大反应时间与培养的细胞数量呈负相关。此外，反应的持续时间受到关键培养基成分快速消耗和/或抑制因子积累的限制，特别是当细胞浓度超过5×10⁵个细胞/培养物时。每天用新鲜培养基培养可部分克服这种影响。分级分离研究表明T细胞和B细胞群体都有需求。通过添加灭活的自体“填充”细胞，可实现PFC产生相对于细胞数量的恒定效率。将讨论这些结果的意义以及微培养系统在详细分析人体抗体反应中的适用性。
