Generation of functional human T cell hybrids.

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

doi:10.1084/jem.154.6.1827

Cited by 47 publications

(6 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast to human BCDF described by others (Muraguchi et al 1981, Okada et al 1983), our hybrids do not induce differentiation in the human B lymphoblastoid line, CESS, while crude supernatants from PHA-stimulated cells demonstrate such activity (data not shown). Our hybrids are not similar to those described by Irigoyen et al (1981) in that PWM stimulation does not enhance the activity of the supernatants or ofthe hybridomas themselves. These comparisons suggest that BCDF may actually consist of several subgroups acting on different B cell subpopulations.…”

Section: Heterogeneity Of Bcdfcontrasting

confidence: 96%

“…However, mitogen-stimulated supernatants or factors derived from cell lines have usually been contaminated with more than one factor, making data interpretation more complex. The generation of human and murine T cell hybridomas has allowed characterization of several of these factors (Okada et al 1983, Mayer et al 1982, Irigoyen et al 1981, Liebson et ai. 1981 as well as describing some new, previously undescribed activities.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Regulation of B Cell Activation and Differentiation with Factors Generated by Human T Cell Hybridomas

Mayer

Kunkel

1984

Immunological Reviews

View full text Add to dashboard Cite

Human T cell hybridomas were generated by several techniques and the supernatants generated were screened for activity on human B cells. Three general activities were noted; B cell proliferation factor ( BCPF ), B cell differentiation factor (BCDF), and an IgA isotype-specific helper factor. BCPF acts on B cells to induce proliferation without differentiation and is distinct from conventional BCGF. This was documented by BCPF 's inability to synergize with anti-mu Ab in a standard BCGF assay ( Muraguchi & Fauci 1982, Howard et al. 1982, Sieckman et al. 1981), as well as its differential effect on a leukemic B cell preparation, when compared with BCGF. A possible schema for BCPF activity is depicted in Figures 3 and 4. In Figure 3, BCPF acts like Ag in vivo or like anti-mu in vitro, pre-activating B cells and rendering them responsive to BCGF. Figure 4 represents what our data depict, that is that BCPF bypasses the response to BCGF and induces cells to proliferate without pre-activation. The difference in the 2 mechanisms may be concentration-dependent and this possibility is currently being evaluated. It is interesting to speculate that T cells in vivo are capable of initiating B cell activation and may account for polyclonal responses seen with some Ag-specific reactions. BCDF(s) act on post-activated B cells (Figure 3) to induce differentiation to Ig-secreting cells. They appear to be heterogeneous and, therefore are capable of inducing varied responses depending on the B cell subpopulation affected. Figure 3 is deliberately complex demonstrating some of the possible as well as documented BCDF activities including polyclonal differentiation and isotype specific activity in IgA committed B cells. We cannot be certain of the frequency of these BCDF-secreting T cells, but the studies of cells from patients with common variable immunodeficiency and chronic lymphocytic leukemia have helped to dissect out these activities. These data would suggest that these BCDF subgroups are important, as deficiencies in one or more subgroups may result in disease.

show abstract

Section: Heterogeneity Of Bcdfcontrasting

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Regulation of B Cell Activation and Differentiation with Factors Generated by Human T Cell Hybridomas

Mayer

Kunkel

1984

Immunological Reviews

View full text Add to dashboard Cite

show abstract

“…J2S 1 induces polyclonai differentiation with increases in IgG and IgM as well as IgA, in striking contrast to J1.3. This hybrid may be similar to that described by Irigoyen et al (5), except that prestimulation of the B cells with PWM was not necessary with J2S1. In addition, two other hybrids were studied that did not induce B cell differentiation, but rather caused active proliferation of resting B cells.…”

Section: Brief Definitive Reportsupporting

confidence: 69%

Human T cell hybridomas secreting factors for IgA-specific help, polyclonal B cell activation, and B cell proliferation.

Mayer¹,

Fu²,

Kunkel³

1982

The Journal of Experimental Medicine

109

View full text Add to dashboard Cite

S u b s t a n t i a l e v i d e n c e has b e e n a c c u m u l a t e d for a m a j o r role of T cells or their factors in B cell proliferation a n d m a t u r a t i o n . R e c e n t reports h a v e suggested that certain T cells m a y be responsible for specific isotype expression on B cells (1-3). T h i s has been most clearly d e m o n s t r a t e d for IgE (1), b u t recent e v i d e n c e has also i m p l i c a t e d such cells in I g G a n d IgA systems (2, 3). T h e r e is also e v i d e n c e that these factors m a y act by i n d u c i n g an Ig class switch (2, 4). t a i n e d w i t h h u m a n T cell h y b r i d o m a s (5-7). T h i s m e t h o d o l o g y was used in our l a b o r a t o r y in an a t t e m p t to o b t a i n different T cell factors i n v o l v e d in B cell a c t i v a t i o n a n d c o m p a r e their effect on isolated n o r m a l B cells a n d m o n o c l o n a l B cell leukemias. Materials and MethodsCell Separation and Culture Conditions. Leukocyte concentrate packs obtained from the New York Blood Center, New York, were used as a source of peripheral blood mononuclear cells (PBMC). Tonsillar tissue was obtained from routine tonsillectomy specimens. Mononuclear cells and T / B separations were prepared as previously described (8). Non-T cells were rosetted twice before use, resulting in <1% O K T 3 + (pan T cell marker; Ortho Pharmaceutical, Raritan, N J) cells in this population. Isolated T cells were cultured in complete medium (8) or subjected to a further rosetting procedure to enrich for OKT-4 + (helper/inducer T cell marker) cells used for fusion. An indirect rosetting technique was used; 40 × 106 T cells were incubated in O K T -8 antibody (suppressor/cytotoxic T cell marker) and rosetted with goat anti-mouse Ig-coated ox erythrocytes (RBC), coated by a chromium chloride technique (8). The resulting interface, after Ficoll-Hypaque gradient centrifugation, was 90-95% O K T -4 + and <1% OKT-8 + by indirect immunofluorescence. The OKT-4 + cells were incubated with concanavalin A (Con A) (Sigma Chemical Co., St. Louis, MO), 10/~g/ml, for 72 h at a cell concentration of 1-2 × 106/ ml. These stimulated cells were washed three times in serum-free medium before use in cell fusion. Establishment of Hypoxanthine Guanine Phosphoribosyl Transferase (HGPR T)-deficient Human T CellLines for Fusion. Jurkat, a human T cell lymphoma line, and KE 37, a human T cell ALL line, were mutagenized with ethylmethane sulfonate at a concentration of 200/~g/ml according to Epstein et al. (9). 80% of treated cells died from this treatment, but after 2 wk in culture, the remaining 20% were _>95% viable. HGPRT-deficient mutants were selected in 6-thioguanine and cloned on soft agar with human 6-thioguanine-resistant fibroblasts

show abstract

“…Another potential explanation of this phenomenon is overgrowth of the cultures by nonfunctional hybrids that are better adapted to tissue culture. Thus, it is possible that early, aggressive cloning is an important step in preserving function of T cell hybridomas, as has been emphasized in a recent study (3).…”

Section: Discussionmentioning

confidence: 97%

“…Functional hybrids derived from both normal and malignant human T cells have been developed. These hybrids have been demonstrated to secrete interleukin 2 (IL-2) 1 (1), killer helper factor (2), and factors that help or suppress immunoglobulin production (3,4). Application of the hybridization technique of Kohler and Milstein (5) to murine T lymphocytes has yielded an even wider variety of factors with immunoregulatory function (6)(7)(8)(9).…”

mentioning

confidence: 99%

Development of a human T-T cell hybridoma secreting B cell growth factor.

Butler¹,

Muraguchi²,

Lane³

et al. 1983

The Journal of Experimental Medicine

View full text Add to dashboard Cite

The success of long-term culture of normal human and murine B cells has been hampered by the limited availability of soluble factors capable of maintaining proliferation of activated B lymphocytes. Previous experiments using various culture-derived supernatants in a human system were unable to separate the activities of B cell growth factor (BCGF) and interleukin 2 (IL-2) by immunochemical means. Thus, purified factors with BCGF activity in the absence of IL-2 activity have not been available for study. In the present study, normal human peripheral blood T cells were fused with the hypoxanthine/aminopterin/thymidine-sensitive human T-leukemic cell line, CEM-6. Supernatants from the resulting hybrid cells were tested for the ability to maintain proliferation of normal human B cells in a recently described assay system for human BCGF. Hybrids demonstrating BCGF activity were cloned by limiting dilution. One hybrid clone, 2B11, continued to support proliferation of B cells in both long-term cultures and 6-d assays at a level significantly above that seen with conventionally produced growth factors. No IL-2 activity was found in the supernatant from hybrid 2B11. The hybridoma supernatant was fractionated by gel filtration, and maximum proliferation of B cells was supported by the 18-20,000 mol wt protein fraction. Thus, a human T-T cell hybridoma that has BCGF activity in the absence of any demonstrable IL-2 activity has been developed. Human T-T cell hybridomas secreting discrete immunoregulatory factors should prove to be powerful tools in dissecting the mechanisms of immunoregulation of human lymphocyte function.

show abstract

Generation of functional human T cell hybrids.

Cited by 47 publications

References 22 publications

Regulation of B Cell Activation and Differentiation with Factors Generated by Human T Cell Hybridomas

Regulation of B Cell Activation and Differentiation with Factors Generated by Human T Cell Hybridomas

Human T cell hybridomas secreting factors for IgA-specific help, polyclonal B cell activation, and B cell proliferation.

Development of a human T-T cell hybridoma secreting B cell growth factor.

Contact Info

Product

Resources

About