Construction of Human T-Cell Hybrids with Helper Function

Irigoyen, Oscar H.; Rizzolo, Philip V.; Thomas, Y; Rogozinski, Linda; Chess, Leonard

doi:10.1007/978-1-4615-9376-8_22

Advances in Experimental Medicine and Biology

1984

DOI: 10.1007/978-1-4615-9376-8_22

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Construction of Human T-Cell Hybrids with Helper Function

Oscar H. Irigoyen

Philip V. Rizzolo

Y Thomas

et al.

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“…In an earlier study we found that HutlO2-B2 cells could be induced with TPA to secrete both IFN-a and IFN-y, though SH9 cells never produced any IFN under the experimental conditions employed (29). In contrast to the reported instability of functional activities of human T hybridomas (14), L265-K and L265-O cells exhibited stable levels ofconstitutive IFN-y secretion. One factor that undoubtedly contributed to this stability was the low propensity of the hybrids to undergo chromosome loss.…”

contrasting

confidence: 62%

“…On the other hand, several hybrids with helper T cell phenotype were obtained by fusing azaguanine-resistant mutants of the human T leukemic line CEM with either unstimulated or lectin/alloantigen-stimulated normal human T lymphocytes. The hybrids either secreted lymphokines, such as TCGF (13), or themselves exhibited nonspecific helper function in vitro for polyclonal induction of antibody production by human B cells (14). In addition, CEM cells treated with RNA and protein synthesis inhibitors were also used for the same purpose, and T hybridomas producing lymphotoxin and migration inhibitory factor were established (15).…”

mentioning

confidence: 99%

“…Recently, it was shown that human T cell hybridomas with functional activities could be constructed from suitable human T leukemic cell lines (12)(13)(14)(15). Fusion of a thymidine kinase-deficient variant of the human T cell line KE37 with peripheral blood lymphocytes (PBL) from a patient with variable agammaglobulinemia resulted in a hybrid showing suppressor/cytotoxic T cell phenotype and producing a soluble factor with suppressive activity on polyclonal immunoglobulin production (12).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Human T cell hybridomas secreting immune interferon.

Le¹,

Vilček²,

Saxinger³

et al. 1982

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

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Human T-cell hybridomas were established by hybridization of concanavalin A-stimulated human peripheral blood lymphocytes with a 6-thioguanine-resistant mutant cell line, designated SH9, derived by irradiation from a cloned human cutaneous T lymphoma line, HutlO2-B2. High levels of interferon (IFN) were demonstrated in the supernatants of hybridoma L265 and its subclones. Whereas no IFN was detected in SH9 cell cultures, up to 1,330 units of IFN per ml were produced spontaneously by the hybrids. On induction with 12-0-tetradecanoylphorbol 13-acetate, IFN synthesis in hybridoma cultures was enhanced 8-to 16-fold. Neutralization with specific antisera and determination of antiviral activities in human and bovine cells showed that the IFN secreted by the hybridomas was immune IFN (IFN-y). Analysis of DNA content, karyotype, and cell surface phenotype, including T cell specific antigens and receptors, confirmed the T cell hybrid nature ofL265 clones. No correlation was. found in the hybridomas between.IFN production and the expression of HTLV, a retrovirus released by HutlO2-B2 and SH9 cells.Somatic cell hybridization techniques have been extensively employed for the production of monoclonal antibodies as well as the characterization of T lymphocytes and their products. Murine AKR T cell lymphoma BW5147 was widely used as a parental line, yielding a variety of mouse hybridomas with diverse T cell functions. In general, the hybridomas could be classified into three main categories: (i) hybridomas mediating specific immune responses, such as induction ofplaque-forming cells (1) and specific tumor cell lysis (2); (ii) hybridomas producing specific T cell factors, including hapten-or carrier-specific helper factors (3, 4), and suppressor factors (5-7); and (iii) hybridomas secreting nonspecific lymphokines-e.g., macrophage migration inhibitory factor (MIF), macrophage activating factor (MAF), T cell-replacing factor (TRF), T cell growth factor [TCGF, interleukin 2 (IL2)], and granulocyte-macrophage colony-stimulating factor (CSF) (8-11). The availability of functional mouse T cell hybrids has provided an important tool for a better understanding of lymphocyte interactions and regulation in the course of immune responses.Recently, it was shown that human T cell hybridomas with functional activities could be constructed from suitable human T leukemic cell lines (12-15). Fusion of a thymidine kinase-deficient variant of the human T cell line KE37 with peripheral blood lymphocytes (PBL) from a patient with variable agammaglobulinemia resulted in a hybrid showing suppressor/cytotoxic T cell phenotype and producing a soluble factor with suppressive activity on polyclonal immunoglobulin production (12). On the other hand, several hybrids with helper T cell phenotype were obtained by fusing azaguanine-resistant mutants of the human T leukemic line CEM with either unstimulated or lectin/alloantigen-stimulated normal human T lymphocytes. The hybrids either secreted lymphokines, such as TCGF (13), or themselves exhibited ...

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confidence: 62%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Human T cell hybridomas secreting immune interferon.

Le¹,

Vilček²,

Saxinger³

et al. 1982

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

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“…These hybridomas may also be a potential source ofmonoclonal lymphokines which regulate a variety of lymphocyte functions, (7). Several reports on the establishment offunctional albeit nonspecific human T hybridomas confirmed this prediction (8)(9)(10).…”

mentioning

confidence: 66%

Antigen-specific human T-cell hybridomas with helper activity.

DeFreitas¹,

Vella²,

Linnenbach³

et al. 1982

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

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Human T cell hybridomas were produced by fusing the hypoxanthine phosphoribosyltransferase-deficient line of the human T cell lymphoma Jurkat with a continuous line of normal human T cells specific for tetanus toxoid (TeT). The hybridomas were selected for their ability to produce interleukin 2 after exposure to TeT on semiautologous monocytes and for their ability to bind to TeT-pulsed semiautologous monocytes. These antigenspecific T hybridomas demonstrated potent helper activity for semiautologous B cells as determined by the production of high levels of anti-TeT antibody in vitro.

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Construction of Human T-Cell Hybrids with Helper Function

Cited by 2 publications

References 35 publications

Human T cell hybridomas secreting immune interferon.

Human T cell hybridomas secreting immune interferon.

Antigen-specific human T-cell hybridomas with helper activity.

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