Establishment of an IL-12-responsive T cell clone: Its characterization and utilization in the quantitation of IL-12 activity
Establishment of an IL-12-responsive T cell clone: Its characterization and utilization in the quantitation of IL-12 activity
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DOI：
10.1002/jlb.61.3.346
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发表时间：
1997-03-01
影响因子：
5.5
通讯作者：
Fuijiwara, H
中科院分区：
文献类型：
--
作者：
Maruo, S;Ahn, HJ;Fuijiwara, H
我们先前证明，终末分化的Thl克隆的增殖主要依赖于由它们与抗原呈递细胞（APC）的相互作用介导的白细胞介素-12（IL-12）-旁分泌机制，而不是IL-α-自分泌机制。（4-86，C57 BL/6来源）在不存在抗原或APC的情况下与重组IL-12（rIL-12）一起培养。一些细胞在仅用rIL-12的培养物中存活数代，并且通过有限稀释，其中一个克隆2D 6对低于1 pg/mL的rIL-12有强烈的增殖反应。该克隆表现出以下表面表型：CD 3（+），T细胞受体（TCR）α β（+），v β 11（+），NK-1.1（-），CD 4（-）CD 8（-），LFA-1（+）、ICAM-1（+）;和CD 28（+）、CD 80（+）、CD 86（+）、CTLA-4（-）。与IL-12的高反应性一致，2D 6细胞也表达IL-12受体（IL-12 R），用rIL-12刺激2D 6细胞后，2D 6细胞表达干扰素γ（IFN-γ）。2D 6克隆对IL-2（IFN-γ）和IL-10 mRNA以及这些细胞因子的产生有应答，（大力）、IL-7（中度）和IL-4（轻度）以及IL-12，然而，使用抗IL-12 mAb的Ab捕获测定使我们能够定量给定样品中所含的IL-12特异性活性，因此，该研究描述了IL-13应答性T细胞克隆的独特特征，并证明了该克隆在定量特异性IL-12活性中的应用。
We previously demonstrated that proliferation of terminally differentiated Thl clones depends primarily on an interleukin-12 (IL-12)-paracrine mechanism mediated by their interactions with antigen-presenting cells (APC) rather than on an IL-a-autocrine mechanism, Such a Thl clone (4-86, C57BL/6 origin) was cultured with recombinant IL-12 (rIL-12) in the absence of either antigen or APC, Some cells survived for several passages of culture with only rIL-12, and by limiting dilution, several clones highly reactive to rIL-18 alone were obtained, One of these clones, designated 2D6, was found to proliferate strongly in response to less than 1 pg/mL of rIL-12, This clone exhibited the following surface phenotypes: CD3(+), T cell receptor (TCR) alpha beta(+), v beta 11(+), NK-1.1(-); CD4(-) CD8(-); LFA-1(+), ICAM-1(+); and CD28(+), CD80(+), CD86(+), CTLA-4(-). In accordance with high responsiveness to IL-12, 2D6 cells were also found to express IL-12 receptor (IL-12R) as detected by incubation with rIL-12 and then staining with anti-IL-12 monoclonal antibody (mAb), Stimulation of 2D6 with rIL-12 resulted in the expression of interferon-gamma (IFN-gamma) and IL-10 mRNAs and production of these cytokines, The 2D6 clone responded to IL-2 (vigorously), IL-7 (moderately), and IL-4 (mildly) in addition to IL-12, However, the Ab capture assay using anti-IL-12 mAb enabled us to quantify IL-12-specific activity contained in a given sample, Thus, this study describes the unique features of the IL-13-responsive T cell clone and demonstrates the utilization of this clone in the quantitation of a specific IL-12 activity.