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Establishment of an IL‐12‐responsive T cell clone: its characterization and utilization in the quantitation of IL‐12 activity
Author(s) -
Maruo Seiji,
Ahn HyunJong,
Yu WenGong,
Tomura Michio,
Wysocka Maria,
Yamamoto Norihiko,
Kobayashi Michiko,
Hamaoka Toshiyuki,
Trinchieri Giorgio,
Fuijiwara Hiromi
Publication year - 1997
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.61.3.346
Subject(s) - biology , clone (java method) , cd28 , microbiology and biotechnology , cd80 , autocrine signalling , cd86 , antigen , monoclonal antibody , t cell , cd8 , interleukin 2 , cell culture , cytokine , receptor , antibody , cytotoxic t cell , cd40 , immunology , immune system , in vitro , biochemistry , dna , genetics
Abstract We previously demonstrated that proliferation of terminally differentiated Thl clones depends primarily on an interleukin‐12 (IL‐12)‐paracrme mechanism mediated by their interactions with antigen‐presenting cells (APC) rather than on an IL‐2‐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‐12 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) αβ + , Vβ11 + , 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‐γ (IFN‐γ) 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‐12‐responsive T cell clone and demonstrates the utilization of this clone in the quantitation of a specific IL‐12 activity. J. Leukoc. Biol . 61: 346–352; 1997.