H-2-restricted antigen binding by a hybridoma clone that produces antigen-specific helper factor.

Lonai, Peter; Puri, Joseph; Hämmerling, Günter J.

doi:10.1073/pnas.78.1.549

Cited by 42 publications

(19 citation statements)

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“…The helper factor was measured by using it to replace syngeneic CGG-primed T cells. Details of the assay have been described previously (16,17).…”

Section: Methodsmentioning

confidence: 99%

“…Clone T85-109-45 was derived by limiting dilution cloning from line T85-109, which is a somatic cell hybrid between NIP-CGG immune B10.BR splenic T cells and BW-5147 AKR lymphoma cells; hence it is homozygous for H-2 k. The fusion and selection of our lines has been described previously (16). The hybridoma cells were grown in RPMI 1640 medium (Gibco Laboratories, Grand Island Biological Co., Grand Island, N.…”

Section: Construction Of the Hybridoma Cellsmentioning

confidence: 99%

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H-2-restricted helper factor secreted by clone hybridoma cells.

Lonai¹,

Puri²,

Bitton³

et al. 1981

The Journal of Experimental Medicine

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Antigen-specific T helper and T suppressor factors are characterized by their affinity to antigen and by determinants controlled by immunoglobulin heavy chain variable region (Vn) 1 and H-2I loci (for reviews, see 1-3). A generally accepted hypothesis suggests that specific T cell factors may be secreted receptors of T cells. It is interesting, however, that whereas the function of most T cells is H-2 restricted, many antigen-specific T cell factors are not H-2 restricted (4-14). It is not clear, therefore, whether some H-2-restricted T cells produce nonrestricted factors, which thus do not completely represent their receptor, or alternatively, the different factors may respectively represent restricted and nonrestricted T cell sets. Because the difference between regular and associative recognition most likely depends on the nature of the receptor, this problem has important structural and genetic implications. Analysis of the question involves studying H-2 restriction of both the cellular receptor and the helper factor from the same T cell clone. Hybridoma clones secreting H-2-restricted (15) or nonrestricted (11-13) suppressor factors have been described. Much less success was achieved, however, in the construction of helper factor-producing hybridomas, and for neither case is there information that would compare factors and receptors.Recently we have described an approach for the preparation of helper hybridoma cultures (16). One of a number of clones was characterized, and it was shown that this clone, T85-109-45, secretes high titers of carrier, chicken gamma globulin (CGG), specific helper factor. It has also been demonstrated that the T85-109-45 factor contains Ia determinants and also determinants shared with the Ig VH framework

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“…The helper factor was measured by using it to replace syngeneic CGG-primed T cells. Details of the assay have been described previously (16,17).…”

Section: Methodsmentioning

confidence: 99%

Section: Construction Of the Hybridoma Cellsmentioning

confidence: 99%

H-2-restricted helper factor secreted by clone hybridoma cells.

Lonai¹,

Puri²,

Bitton³

et al. 1981

The Journal of Experimental Medicine

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“…These were described previously (5,6,12). Briefly, anti-(4-hydroxy-5-iodo-3-nitrophenyl)acetyl (NIP) antibodies were detected by counting NIPspecific plaques produced by NIP-ovalbumin (OVA)-primed spleen cells in the presence of hybridoma-derived helper factors or CGG-specific syngeneic T cells.…”

Section: Methodsmentioning

confidence: 99%

“…Affinity Chromatography. Affinity chromatography on antibody-Sepharose and on normal spleen cells has been described previously (5,6,12). Table II demonstrates that the supernatant of this clone, which derives from the fusion of an H-2 b T cell with an H-2 k lymphoma, was active only towards B cell sources expressing H-2 k or H-2 b gene products but not towards those that express H-2 q orf (9-2 q andf were chosen as foreign H-2 types because they contain the least shared Ia specifieities with haplotypes b and k [18]).…”

Section: Designation Of the Hybridoma Clonesmentioning

confidence: 99%

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Two separate genes regulate self-Ia and carrier recognition in H-2-restricted helper factors secreted by hybridoma cells.

Lonai

Bitton

Savelkoul

et al. 1981

The Journal of Experimental Medicine

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Since the discovery that T cells recognize antigen in conjunction with major histocornpatibility complex (MHC) I products, most workers have assumed that the problem as to whether this is due to two receptors recognizing two determinants (dual recognition) or to one receptor recognizing a neoantigen (altered self) could be solved by means outside classical cellular immunological methods (1, 2). In the past few years significant advances were made in T cell culture methods. Today these cells can be grown as growth factor-dependent continuous lines (3) or as somatic cell hybrids (4). These clonable lines represent homogeneous sources of the T cell receptor. The hybridoma technology used in this context to "immortalize" T cells is a somatic genetic approach, suitable for analyzing whether a trait is controlled by one or more loci. This question appears to be at the center of the altered self/dual recognition controversy. As an initial step in solving this problem, we studied H-2-restricted carrier-specific helper factors secreted by H-2 heterologous T cell hybrids.We recently reported (5) the isolation of a number of 11-2 homologous and heterologous helper factor producing T cell hybrids. One of these, deriving from H-2 homologous fusion partners, was described in detail (6). The factor of this clone, similarly to T cell factors and receptors, contained determinants shared with immunoglobulin heavy chain variable regions (IgVH) (6-11) and also with Ia antigens. Significantly, its helper activity was H-2 restricted (6). In our experiments presented here, we were guided by the idea that the factors secreted by the H-2 heterologous clones, if they are also H-2 restricted, could be used to study whether both H-2 genomes of the fusion partners are involved in H-2-restricted helper factors. Insofar as an H-2-restricted T cell factor represents the receptor of the cells secreting it, such an approach could be helpful in understanding H-2 restriction in general.* Address correspondence to P. Lonai, Department of Chemical Immunology, The Weizmann Institute of Science, Rehovot 76100, Israel.:~ In partial fulfillment of the Master of Science thesis requirements of the Agricultural University of Wageningen, The Netherlands.Abbreviatwns used in this paper: CGG, chicken gamma globulin; Ca, immunoglobulin heavy chain constant region; HGG, human gamma globulin; IgVn or Vn, immunoglobulin heavy chain variable region; MHC, major histocompatibility complex; NIP, (4-hydroxy-5-iodo-3-nitrophenyl) acetyt; NP, (4-hydroxy-3-nitrophenyi) acety[; OVA, ovaibumin; PFC, plaque-forming cells; V, and Vx, variable region of~ or .~ fight chains, respectively. 1910J. Exp. MED.

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Differentiation of Function among Antigen‐Presenting Cells

Humphrey

1981

Ciba Foundation Symposium 84 ‐ Microenvironments in Haemopoietic and Lymphoid Differentiation

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H-2-restricted antigen binding by a hybridoma clone that produces antigen-specific helper factor.

Cited by 42 publications

References 18 publications

H-2-restricted helper factor secreted by clone hybridoma cells.

H-2-restricted helper factor secreted by clone hybridoma cells.

Two separate genes regulate self-Ia and carrier recognition in H-2-restricted helper factors secreted by hybridoma cells.

Differentiation of Function among Antigen‐Presenting Cells

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