Moretta A, Poggi A, Olive D, Bottino C, Fortis C, Pantaleo G, Moretta L
Proc Natl Acad Sci U S A. 1987 Mar;84(6):1654-8. doi: 10.1073/pnas.84.6.1654.
A clone of the interleukin 2-producing Jurkat leukemia cell line termed JA3 (surface phenotype, T3+, Ti+, T44+, T11+, T40+) has been used to induce and select cell variants lacking surface molecules involved in T-cell activation. Following 200 rad of gamma-radiation (1 rad = 0.01 Gy), cells were treated with monoclonal antibodies (mAbs) directed to T3, Ti, T44, or T11 antigen and complement. After growth of the residual cells in culture, "negative" cells were cloned under limiting conditions. Depending on the specificity of the mAb used for the immunoselection, three groups of variants were obtained. (i) The use of mAbs directed to T3 or Ti resulted in cell variants that expressed the T3 Ti- T44+ Leu1+ T11+ T40+ 4F2+ HLA class I+ surface phenotype. (ii) Immunoselection with anti-T44 mAb resulted in 2 variants that shared the T3- Ti- T44- Leu1- T11+ T40+ 4F2+ HLA class I+ phenotype. (iii) Cell treatment with anti-T11 mAb resulted in 15 variants characterized by the lack of T11 antigen expression and of all the other T-cell-specific surface antigens. Therefore, it appears that the different sets of JA3 cell variants, like T cells at discrete stages of intrathymic differentiation, may follow a coordinated expression of surface differentiation antigens. Analysis of the functional responsiveness of the three distinct groups of JA3 cell variants to different stimuli showed that all produced interleukin 2 in response to A23187 calcium ionophore plus phorbol 12-myristate 13-acetate. The first group of variants (T3- Ti-) did not respond to stimulation with anti-T3, anti-Ti, or anti-T44 mAbs. Eight of 9 did not respond to phytohemagglutinin either; however, all responded to appropriate stimulatory combinations of anti-T11 mAbs (and to calcium ionophore). The second group of variants (T3-, Ti-, T44-, T11+), similar to the first group, did not respond to anti-T3, anti-Ti, anti-T44 mAbs, and phytohemagglutinin, but they were fully responsive to anti-T11 mAb. The last group of variants (lacking all the T-cell-specific surface antigens) only responded to calcium ionophore A23187.
一种名为JA3的产生白细胞介素2的Jurkat白血病细胞系克隆株（表面表型为T3 +、Ti +、T44 +、T11 +、T40 +）已被用于诱导和筛选缺乏参与T细胞活化的表面分子的细胞变体。在200拉德的γ射线辐射（1拉德 = 0.01戈瑞）后，细胞用针对T3、Ti、T44或T11抗原的单克隆抗体（mAb）和补体进行处理。在培养中残留细胞生长后，在有限条件下克隆“阴性”细胞。根据用于免疫筛选的mAb的特异性，获得了三组变体。（i）使用针对T3或Ti的mAb导致细胞变体表达T3 - Ti - T44 + Leu1 + T11 + T40 + 4F2 + HLA I类 + 表面表型。（ii）用抗T44 mAb进行免疫筛选产生了2个变体，它们具有T3 - Ti - T44 - Leu1 - T11 + T40 + 4F2 + HLA I类 + 表型。（iii）用抗T11 mAb处理细胞产生了15个变体，其特征是缺乏T11抗原表达以及所有其他T细胞特异性表面抗原。因此，似乎不同组的JA3细胞变体，就像胸腺内分化不同阶段的T细胞一样，可能遵循表面分化抗原的协调表达。对三组不同的JA3细胞变体对不同刺激的功能反应性分析表明，所有变体在响应A23187钙离子载体加佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯时都会产生白细胞介素2。第一组变体（T3 - Ti -）对用抗T3、抗Ti或抗T44 mAb刺激无反应。9个中有8个对植物血凝素也无反应；然而，所有变体都对适当的抗T11 mAb刺激组合（以及对钙离子载体）有反应。第二组变体（T3 -、Ti -、T44 -、T11 +）与第一组类似，对抗T3、抗Ti、抗T44 mAb和植物血凝素无反应，但它们对抗T11 mAb完全有反应。最后一组变体（缺乏所有T细胞特异性表面抗原）仅对钙离子载体A23187有反应。
