Reactivity of hybridomas derived from T cells activated in vivo during graft-versus-host disease

Ramona R. Leibnitz, Peter E. Lipsky, Dwain L Thiele

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Abstract

To examine the specificity of T helper cells activated during murine graft-vs-host disease (GVHD), T cell hybridomas from GVHD spleens and livers were generated and analyzed. CTL-depleted C57BL/6 (B6) donor cells were injected into irradiated (B6 x bm12)F1 or (bm1 x bm12)F1 recipient mice. Five or fourteen days later, cells from livers and spleens were fused directly with the TCR-deficient (αβ)- BW5147 thymoma line. The in vivo- activated T cells produced hybridomas as efficiently as either T cells activated in a primary mixed lymphocyte reaction or expanded in vitro after isolation from GVHD mice. Overall, 91% (396 of 437) of hybridomas generated from GVHD animals responded to immobilized anti-CD3 and 56% (220 of 396) of these hybridomas responded specifically to APC expressing host bm1 or bm12 alloantigens. More than 80% of bm12-specific hybridomas expressed CD4; all (53 of 53) of the bm12-specific hybridomas tested reacted to homozygous bm12 APC. Of the alloreactive T hybridomas generated from B6→(bm1 x bm12)F1 GVHD mice, 7% responded to bm1 APC. Five bm1-specific hybridomas were analyzed further. One CD4+ hybridoma recognized a bm1 peptide presented by self I- Ab and was blocked by anti-Ia Ab; the other four hybridomas, two of which also expressed CD4, responded to transfected L cells expressing H-2K(bm1) and were not inhibited by anti-Ia Ab. These results indicate that a high percentage of CD4+ T hybridomas generated from freshly isolated T cells activated in vivo during GVHD are specific for host MHC class II or class I alloantigens.

Original languageEnglish (US)
Pages (from-to)4959-4968
Number of pages10
JournalJournal of Immunology
Volume153
Issue number11
StatePublished - Dec 1 1994

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Hybridomas
Graft vs Host Disease
T-Lymphocytes
Isoantigens
Spleen
Mixed Lymphocyte Culture Test
Thymoma
Liver
Helper-Inducer T-Lymphocytes
Peptides

ASJC Scopus subject areas

  • Immunology

Cite this

Reactivity of hybridomas derived from T cells activated in vivo during graft-versus-host disease. / Leibnitz, Ramona R.; Lipsky, Peter E.; Thiele, Dwain L.

In: Journal of Immunology, Vol. 153, No. 11, 01.12.1994, p. 4959-4968.

Research output: Contribution to journalArticle

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abstract = "To examine the specificity of T helper cells activated during murine graft-vs-host disease (GVHD), T cell hybridomas from GVHD spleens and livers were generated and analyzed. CTL-depleted C57BL/6 (B6) donor cells were injected into irradiated (B6 x bm12)F1 or (bm1 x bm12)F1 recipient mice. Five or fourteen days later, cells from livers and spleens were fused directly with the TCR-deficient (αβ)- BW5147 thymoma line. The in vivo- activated T cells produced hybridomas as efficiently as either T cells activated in a primary mixed lymphocyte reaction or expanded in vitro after isolation from GVHD mice. Overall, 91{\%} (396 of 437) of hybridomas generated from GVHD animals responded to immobilized anti-CD3 and 56{\%} (220 of 396) of these hybridomas responded specifically to APC expressing host bm1 or bm12 alloantigens. More than 80{\%} of bm12-specific hybridomas expressed CD4; all (53 of 53) of the bm12-specific hybridomas tested reacted to homozygous bm12 APC. Of the alloreactive T hybridomas generated from B6→(bm1 x bm12)F1 GVHD mice, 7{\%} responded to bm1 APC. Five bm1-specific hybridomas were analyzed further. One CD4+ hybridoma recognized a bm1 peptide presented by self I- Ab and was blocked by anti-Ia Ab; the other four hybridomas, two of which also expressed CD4, responded to transfected L cells expressing H-2K(bm1) and were not inhibited by anti-Ia Ab. These results indicate that a high percentage of CD4+ T hybridomas generated from freshly isolated T cells activated in vivo during GVHD are specific for host MHC class II or class I alloantigens.",
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