Lamb J R, Eckels D D, Lake P, Johnson A H, Hartzman R J, Woody J N
J Immunol. 1982 Jan;128(1):233-8.
Human peripheral blood lymphocytes from an HLA-Dw1,3 individual were primed in vitro with influenza A virus (A/Texas/1-77/x-49) and subsequently cloned by limiting dilution in TCGF. Of the 96 TLCs originally obtained, nine were characterized in detail. TLCs were antigen specific, responding to influenza A virus, not to influenza B, TGAL, GAT, tetanus toxoid, or KLH, and only when antigen was presented by cells unable to form rosettes with AET-treated SRBC. Presentation of antigen by unseparated PBL often resulted in significant "back stimulation," probably via production of growth factors. The MHC requirements for the induction of TLC proliferation were analyzed. Of four representative clones analyzed, three required Dw1;DR1 compatibility for successful presentation of viral antigens by a panel of antigen-presenting cells. In contrast, one TLC showed an unusual pattern of response that could not be correlated to a particular HLA haplotype. Monoclonal anti-T cell antibody analysis of the surface phenotype of two TLCs maintained in continuous culture for 5 mo indicated that they were OKT3+, 4+, and 8-, consistent with an inducer/helper phenotype. To confirm the clonal nature of TLCs, data on the functional properties of TLC subclones are also presented.
从一名HLA-Dw1,3个体获取的人外周血淋巴细胞，在体外经甲型流感病毒（A/德州/1-77/x-49）致敏，随后在TCGF中通过有限稀释法进行克隆。最初获得的96个TLC中，对9个进行了详细表征。TLC具有抗原特异性，对甲型流感病毒有反应，对乙型流感病毒、TGAL、GAT、破伤风类毒素或KLH无反应，且仅当抗原由不能与AET处理的SRBC形成花环的细胞呈递时才有反应。未分离的PBL呈递抗原通常会导致显著的“反向刺激”，可能是通过生长因子的产生。分析了诱导TLC增殖的MHC要求。在分析的4个代表性克隆中，3个克隆需要Dw1;DR1相容性，以便一组抗原呈递细胞成功呈递病毒抗原。相比之下，一个TLC表现出一种异常的反应模式，无法与特定的HLA单倍型相关联。对连续培养5个月的两个TLC的表面表型进行单克隆抗T细胞抗体分析表明，它们为OKT3 +、4 +和8 -，与诱导/辅助表型一致。为了证实TLC的克隆性质，还给出了TLC亚克隆功能特性的数据。
