Gerhard W, Hackett C, Melchers F
J Immunol. 1983 May;130(5):2379-85.
A T cell line was established in long-term tissue culture from spleen cells of BALB/c mice immunized with influenza virus PR8 (A/PR/8/34(H1N1)) by continuous restimulation with PR8 virus and syngeneic x-irradiated spleen cells. This T cell line and clones derived from it have now been propagated for over 1 yr "in vitro". The T cell line and the clones, upon stimulation with Con A in the absence of antigen and irradiated spleen cells, as upon stimulation in the presence of antigen and adherent cells, produce T cell growth factors, B cell replication and maturation factors, and colony-stimulating factors, as tested by restimulation of proliferation of a clone of cytolytic murine T cells, of lipopolysaccharide-activated B cell blasts, and by macrophage/granulocyte colony formation of murine bone marrow cells, respectively. The T cell line and the clones help syngeneic B cells in the presence of syngeneic irradiated spleen cells and PR8 virus to proliferate and mature into clones of cells secreting virus-specific antibodies. In the presence of PR8 virus and of a bystander antigen, such as sheep red cells, B cells specific for this bystander antigen are also induced. Adoptive transfer of the T cells i.v. into syngeneic nu/nu BALB/c mice enable the recipient mice to produce, upon challenge with PR8, virus-specific antibodies and to clear virus infection of the lung. The virus-specific T cell line and the clones are restricted by H-2 antigens of the d-haplotype, probably by Iad, and recognize a determinant on the hemagglutinin (HA) molecule of PR8 virus. Fifty to 100 fmol of virus-associated or free HA suffice for stimulation as measured by proliferation assays. The stimulating determinant is present on the HA of all natural virus isolates of the H1 subtype, is absent from virus isolates of the H2 and H3 subtypes, and is abolished if glutamic acid at position 115 of the HA1 polypeptide of PR8 is replaced by lysine.
用流感病毒PR8（A/PR/8/34(H1N1)）免疫BALB/c小鼠，取其脾细胞经PR8病毒和同基因X射线照射的脾细胞连续再刺激，在长期组织培养中建立了一个T细胞系。这个T细胞系及其衍生的克隆现已在体外传代培养超过1年。该T细胞系和克隆在无抗原和照射脾细胞存在的情况下用刀豆球蛋白A刺激时，以及在有抗原和贴壁细胞存在的情况下刺激时，分别通过对溶细胞性小鼠T细胞克隆、脂多糖激活的B细胞母细胞增殖的再刺激，以及对小鼠骨髓细胞巨噬细胞/粒细胞集落形成的检测，产生T细胞生长因子、B细胞复制和成熟因子以及集落刺激因子。该T细胞系和克隆在同基因照射的脾细胞和PR8病毒存在的情况下帮助同基因B细胞增殖并成熟为分泌病毒特异性抗体的细胞克隆。在PR8病毒和旁观者抗原（如绵羊红细胞）存在的情况下，也可诱导针对该旁观者抗原的B细胞。将T细胞静脉内移植到同基因无胸腺BALB/c小鼠中，使受体小鼠在受到PR8攻击时能够产生病毒特异性抗体并清除肺部的病毒感染。该病毒特异性T细胞系和克隆受d单倍型的H-2抗原限制，可能受Iad限制，并识别PR8病毒血凝素（HA）分子上的一个决定簇。通过增殖试验测定，50至100飞摩尔的病毒相关或游离HA足以产生刺激作用。刺激决定簇存在于H1亚型的所有天然病毒分离株的HA上，不存在于H2和H3亚型的病毒分离株上，如果PR8的HA1多肽第115位的谷氨酸被赖氨酸取代，则该决定簇被消除。
