Construction of T cell hybridomas secreting allogeneic effect factor.

Katz, David H.; Bechtold, Thomas

doi:10.1084/jem.152.4.956

Cited by 20 publications

(4 citation statements)

References 27 publications

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“…Infected mice were allowed to resolve primary infection, and were re-challenged two months later. The spleen and lymph nodes were collected one-week post-secondary challenge, and single-cell suspensions were stimulated ex vivo with reticulate body (RB)-enriched Nigg (1μg/mL) (31) for 5 days prior to fusion with murine BW5147 T cell lymphoma cells (32) in 50% PEG solution. Fused cells were cultured in HAT medium for an additional 7 to 9 days.…”

Section: Methodsmentioning

confidence: 99%

A Chlamydia-Specific TCR-Transgenic Mouse Demonstrates Th1 Polyfunctionality with Enhanced Effector Function

Poston

Girardi

et al. 2017

The Journal of Immunology

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Chlamydia is responsible for millions of new infections annually, and current efforts focus on understanding cellular immunity for targeted vaccine development. The Chlamydia-specific CD4 T cell response is characterized by the production of IFNγ, and polyfunctional Th1 responses are associated with enhanced protection. A major limitation in studying these responses is the paucity of tools available for detection, quantification, and characterization of polyfunctional, antigen-specific T cells. We addressed this problem by developing a TCR transgenic mouse with CD4 T cells that respond to a common antigen in Chlamydia muridarum and Chlamydia trachomatis. Using an adoptive transfer approach, we show that naïve transgenic CD4 T cells become activated, proliferate, migrate to the infected tissue, and acquire a polyfunctional Th1 phenotype in infected mice. Polyfunctional Tg Th1 effectors demonstrated enhanced IFNγ production compared to polyclonal cells, protected immune deficient mice against lethality, mediated bacterial clearance, and orchestrated an anamnestic response. Adoptive transfer of Chlamydia-specific CD4 TCR Tg T cells with polyfunctional capacity offers a powerful approach for analysis of protective effector and memory responses against chlamydial infection, and demonstrates that an effective monoclonal CD4 T cell response may successfully guide subunit vaccination strategies.

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Section: Methodsmentioning

confidence: 99%

A Chlamydia-Specific TCR-Transgenic Mouse Demonstrates Th1 Polyfunctionality with Enhanced Effector Function

Poston

Girardi

et al. 2017

The Journal of Immunology

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“…Derivation of the Tcell hybridoma clone 24/G1 has been described (6,8,9). Briefly, alloantigen-activated T-cell blasts were fused with T lymphoma BW5147.…”

Section: Methodsmentioning

confidence: 99%

Macrophage activation: priming activity from a T-cell hybridoma is attributable to interferon-gamma.

Pace¹,

Russell²,

Schreiber³

et al. 1983

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

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165

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Antiviral and macrophage-priming activities in the supernatant medium of a subelone of a concanavalin A-stimulated mouse T-cell hybridoma were investigated. The two activities were associated with a molecular weight of approximately 50,000 and could not be separated by various approaches. Both activities were eliminated by a highly specific neutralizing antibody against mouse interferon-v, but not by antibody against interferon-a and -(P. The ratio of-priming to antiviral activity in the hybridoma culture supernate was indistinguishable from the ratio obtained with mouse interferon-yprepared by recombinant DNA technology. It was concluded from these data that the priming activity in hybridoma culture supernates was attributable to interferon-and that this mediator is one form of the lymphokine macrophage-activating factor. Interferon-y was greater than 800 times more efficient at priming mouse macrophages for tumor cell killing than was a mixture of interferon-a and -(3. This finding contributes to growing awareness that type II interferon may have greater immunoregulatory potential than type I interferons.Macrophages can be activated to kill tumor cells in vitro by a nonspecific, extracellular mechanism that may be important in host defense against neoplastic cells in vivo. An important goal, therefore, is acquisition of better understanding of how the process of activation is regulated. Recently, it has been shown that under defined conditions the lymphokine macrophage-activating factor (MAF) does not render macrophages fully cytolytic (1-5). Instead, it heightens their responsiveness to a second signal(s) that then triggers the expression of killing-i.e., MAF "primes" macrophages to respond to a second, triggering stimulus.A cloned hybridoma (24/Gl) has been recognized that produces MAF, identified by its ability to prime mouse macrophages for tumor cell killing (6). The hybridoma-derived MAF was shown by various biochemical and functional criteria to be indistinguishable from conventionally prepared MAF (6). These supernates also contained antiviral activity attributed to interferon (IFN)-y.We report here that the antiviral and macrophage-priming activities produced by cells of a subclone of 24/Gl are not dissociable by various approaches. These results, together with data obtained by using recombinant mouse IFN-y, support a conclusion that priming activity produced by this hybridoma is attributable to IFN-y. We also report here that IFN-y is much more potent as a macrophage priming agent than type I interferon is. MATERIALS AND METHODSMice. Male C3H/HeN mice were obtained from Charles River Breeding Laboratories (Kingston, NY) and used at 6-9 weeks of age.

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“…The first is the establishment of long-term cultures of normal antigen-specific T cells through the use of repeated in vitro antigen stimulation and/or the growth factor, interleukin 2 (IL-2; formerly, T cell growth factor) (27-30). The second is the production of immortal T cell hybrids between normal antigenspecific T cells and T cell tumor lines (31)(32)(33)(34)(35)(36). In the present study, we combined these two methods to produce inducible, antigen-specific, H-2-restricted T cell hy-bridomas.…”

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confidence: 99%

Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition

Kappler¹,

Skidmore²,

White³

et al. 1981

The Journal of Experimental Medicine

971

322

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The characterization of the antigen-recognizing structure on the various classes of T cells has been a difficult problem for immunologists. Three aspects of this problem are particularly controversial: first, whether the genes that code for the variable portion of immunoglobulin also code for the antigen-recognizing portion of the receptor on all classes of T cells (1-4); second, whether T cell receptors specific for allo-major histocompatibility complex (MHC) 1 products are unique or are included within the population of receptors for conventional antigens (5-10); and third, how T cells, particularly those of the cytotoxic and helper classes, are able to demonstrate an apparent dual specificity for both a conventional antigen and a product of the MHC such that they function only when confronted with both the appropriate antigen and MHC product (11-16). Various models have been proposed to explain this phenomenon. Some have proposed that antigen and MHC are recognized independently via two receptors on the T cell (17-23). Others have proposed various forms of dependent recognition, involving, for example, a single receptor on the T cell, and/or a physical interaction between antigen and MHC products in antigenpresenting cells (24)(25)(26). A number of experimental results have been used by various investigators to argue for one or the other of these models, but there is little evidence that distinguishes the various possibilities.A direct attack on these questions has been hampered by the lack of suitable sources of clonal T cell lines analogous to the B cell myelomas. Recently, two techniques have offered promise in this regard. The first is the establishment of long-term cultures of normal antigen-specific T cells through the use of repeated in vitro antigen stimulation and/or the growth factor, interleukin 2 (IL-2; formerly, T cell growth factor) (27-30). The second is the production of immortal T cell hybrids between normal antigenspecific T cells and T cell tumor lines (31-36). In the present study, we combined these two methods to produce inducible, antigen-specific, H-2-restricted T cell hy-*

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Construction of T cell hybridomas secreting allogeneic effect factor.

Cited by 20 publications

References 27 publications

A Chlamydia-Specific TCR-Transgenic Mouse Demonstrates Th1 Polyfunctionality with Enhanced Effector Function

A Chlamydia-Specific TCR-Transgenic Mouse Demonstrates Th1 Polyfunctionality with Enhanced Effector Function

Macrophage activation: priming activity from a T-cell hybridoma is attributable to interferon-gamma.

Antigen-inducible, H-2-restricted, interleukin-2-producing T cell hybridomas. Lack of independent antigen and H-2 recognition

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