Source and Functional Characteristics of the Mitogenic Factor Inducing Continued Proliferation of Human T Cells

Bonnard, Guy D.; Yasaka, Ken; Jacobson, Dennis; Fine, Robert L.; Morgan, D. A. L.; Herberman, Ronald B.

doi:10.1016/b978-0-12-569650-0.50074-7

Cited by 8 publications

(10 citation statements)

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“…The input cpm for each determination varied from 400 to 800 cpm. The following antibodies were used: anti-Thy-i (Jlj), anti-Lyt-1 (53.7.3), anti-Lyt-2 (53.6.7), anti-IgM (Bet-2), and anti-rat K (MAR 18.5 (7,8) or binding radiolabeled IL-2 (9). Second,.7D4 inhibited IL-2-driven proliferation of HT2 and CTLL cells, two cell lines that are widely used as indicators for IL-2 and that do not appear to require any other exogenous lymphokines for continued growth.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex.

Malek

Robb

Shevach

1983

Proc. Natl. Acad. Sci. U.S.A.

293

133

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Xenogeneic monoclonal antibodies were prepared to the murine interleukin 2 (1L-2)-dependent HT2 cell line. One rat IgM monoclonal antibody (7D4) was identified that inhibited proliferation of the HT2 cells and of IL-2-dependent CTLL cells in the presence of crude rat IL-2 as well as of purified human L-2. The level of inhibition was dependent on both antibody and IL-2 concentration. Cell distribution studies using a fluorescenceactivated cell sorter showed that the antigen identified by 7D4 is expressed at a high density on, HT2 cells and on concanavalin A (Con A)-induced T-cell blasts and at a substantially lower density on lipopolysaccharide-induced B-cell blasts; 7D4 (5) and CTLL (6)-that solely depend on an exogenous source of IL-2 for growth has provided a definitive assay cell population to assess the presence of IL-2 in crude culture fluids containing many biologically active molecules and has greatly aided the purification and biochemical characterization of this lymphokine. IL-2 exerts its growth-promoting properties after interaction with specific membrane binding sites (IL-2 receptors) expressed on activated, but not on resting, T cells (7-9). After IL-2 associates with its receptor, it is internalized and undergoes lysosomal degradation (9). The relationship between the degradation and biological function of IL-2 is unclear. Further delineation of the molecular structures and pathways involved in the IL-2 growth mechanism should be facilitated as monoclonal antibodies reactive with the specific components of the hormone-receptor complex become available. Recently, monoclonal antibodies to IL-2 (10, 11) and the human IL-2 receptor (12) have been described. In the present paper, we report the characterization of a rat monoclonal antibody (7D4) that is reactive with an epitope either on the murine IL-2 receptor distal to the IL-2 hormone binding site or to another molecule physically complexed with the IL-2 receptor.MATERIALS AND METHODS Animals. Inbred strain 2 guinea pigs, C57BL/6 and BALB/ c mice, and Lewis rats were obtained from the Animal Production Section, Division of Research Services, National Institutes of Health (Bethesda, MD).Cell Lines. The properties of the murine IL-2-dependent HT2 cells and of CTLL cells have been described by Watson (5) and by Baker et al. (6), respectively. The nonsecretor hybridoma cell line SP2/0-Ag-14 (SP2/0) was used for all fusions.Antisera and Monoclonal Antibodies. Heterologous classspecific antisera to rat IgM, IgG, and IgA were obtained from Cappel Laboratories (Cochranville, PA) and used to determine the isotypes of rat monoclonal antibodies by Ouchterlony analysis of concentrated culture supernatant fluids. The following culture supernatants containing rat monoclonal antibodies to murine lymphocyte surface antigens were used: 53.7.3 (anti-Lyt-1) and 53.6.7 (anti-Lyt-2), developed by Ledbetter and Herzenberg (13) The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "adv...

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Section: Resultsmentioning

confidence: 99%

“…IL-2 exerts its growth-promoting properties after interaction with specific membrane binding sites (IL-2 receptors) expressed on activated, but not on resting, T cells (7)(8)(9). After IL-2 associates with its receptor, it is internalized and undergoes lysosomal degradation (9).…”

mentioning

confidence: 99%

Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex.

Malek

Robb

Shevach

1983

Proc. Natl. Acad. Sci. U.S.A.

293

133

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“…The controls were cultures of PBMNC that had not been primed to the antigen or that were seeded in the agar layer without antigen or with an irrelevant antigen. The cultures were incubated for [3][4][5][6] days during which the development of colonies was observed through an inverted microscope. The number of colonies was estimated by counting colonies of approximately 50 or more cells from six culture plates.…”

Section: Methodsmentioning

confidence: 99%

“…Early studies by Ben-Sasson et at (3) demonstrated that enriched populations of proliferating guinea pig T lymphocytes could be obtained by repeated stimulation and selection on antigen-pulsed macrophages. With the discovery and characterization of T-cell growth factor (TCGF) (4)(5)(6)(7), it has become possible to maintain long-term cultures of antigen-specific (8)(9)(10)(11) and alloreactive (12)(13)(14)(15)(16) murine and human T lymphocytes in vitro. More recently, several laboratories have succeeded in cloning murine or human cells from long-term lines with the help of TCGF either by limiting-dilution techniques (16)(17)(18)(19) or in soft agar (20,21).…”

mentioning

confidence: 99%

Antigen-specific human T-cell clones: development of clones requiring HLA-DR-compatible presenting cells for stimulation in presence of antigen.

Sredni¹,

Volkman²,

Schwartz³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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A method for cloning soluble antigen-specific proliferating human T lymphocytes directly from peripheral blood cells of individuals recently primed with the antigen is described. The soluble antigen was keyhole limpet hemocyanin. After expansion in liquid culture, these cells were shown to be antigen specific and to require HLA-DR-histocompatible presenting cells. Re Cell Membrane Markers. Cloned cells were characterized for sheep erythrocyte receptors by a rosette procedure as described (24). In addition, assays for the presence of surface membrane immunoglobulin and complement receptors were performed on the cloned cells as described (24). Other membrane antigens were characterized by indirect immunofluorescence with two mouse monoclonal antibodies. The 3A1 antibody (25) delineates a major peripheral blood T-cell subset; the presence of Ia molecules was assessed by using a heterologous rabbit anti-p23,30 antiserum (gift of Dean Mann). Indirect immunofluorescence to detect cells binding these monoclonal antibodies was performed as described (25).TCGF. TCGF was produced as described by Morgan et at (5) and Ruscetti et al (4) 1858The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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“…The ultimate effects of IL-2, induction of DNA synthesis and mitosis, occur after a sequence of events initiated by binding of IL-2 to a specific cellular receptor; the IL-2 receptor is expressed only on antigen-or lectin-activated lymphocytes but not on quiescent cells [2]. We have previously shown that IL-2 acts on the restricted process of a cell cycle which occurs 3-5 h before the beginning of DNA synthesis [3], although the molecular mechanism by which IL-2 produces its effect on cell multiplication has not been identified.…”

Section: Introductionmentioning

confidence: 99%

Interleukin 2 induces rapid phosphorylation of cellular proteins in murine T‐lymphocytes

Kohno¹,

Kuwata²,

Namba³

et al. 1986

FEBS Letters

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When quiescent murine T‐lymphocyte cells were stimulated by the addition of interleukin 2 (IL‐2), they reinitiated DNA synthesis after a lag period of 5 h. Under these conditions, rapid but transient phosphorylation of two cellular proteins with M r, values of 27000 and 26000 was detected; maximal phosphorylation occurred within 10–15 min after the addition of IL‐2. The protein of M r, 27 000 contained phosphoserine, while the protein of M r 26000 contained phosphothreonine.

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Source and Functional Characteristics of the Mitogenic Factor Inducing Continued Proliferation of Human T Cells

Cited by 8 publications

References 1 publication

Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex.

Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex.

Antigen-specific human T-cell clones: development of clones requiring HLA-DR-compatible presenting cells for stimulation in presence of antigen.

Interleukin 2 induces rapid phosphorylation of cellular proteins in murine T‐lymphocytes

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