Expression of the T Cell Gamma Gene by a Functionally Defined Human T Cell Clone Characterization at DNA, RNA, and Cell Membrane Level
V. DAVID
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorG. LECA
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorE. VILMER
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorP. GUGLIELMI
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorS. CHOUAIB
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorJ. KANELLOPOULOS
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorF. SIGAUX
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorCorresponding Author
A. BENSUSSAN
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
A. Bensussan, PhD, INSERM U93, Hôpital Saint Louis, I rue Claude Vellefaux, 75474 Paris Cedex 10, FranceSearch for more papers by this authorV. DAVID
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorG. LECA
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorE. VILMER
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorP. GUGLIELMI
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorS. CHOUAIB
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorJ. KANELLOPOULOS
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorF. SIGAUX
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
Search for more papers by this authorCorresponding Author
A. BENSUSSAN
INSERM U93, Hôpital Saint-Louis, INSERM U23 Hôpital Saint-Antoine, INSERM U108, Hôpital Saint-Louis, INSERM U277 Institut Pasteur, and INSERM U301 and Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France
A. Bensussan, PhD, INSERM U93, Hôpital Saint Louis, I rue Claude Vellefaux, 75474 Paris Cedex 10, FranceSearch for more papers by this authorAbstract
In the present study we describe one CD2+CD3+ clone termed DS6 which expressed neither CD4 nor CD8 differentiation antigens and failed to react with WT31, a monoclonal antibody directed against the T cell antigen receptor α/β helerodimer. This clone was isolated from peripheral blood T lymphocytes of a patient with a prolonged immunodeficiency after allogencic bone marrow transplantation. Normal -sized T cell γ gene transcripts were detected in DS6 by northern analysis, whereas no mature β or α chain mRNA were found The rearrangement of TCRβ chain genes and T cell γ genes was analysed. While in DS6, TCRβ chain genes remain in germinal configuration, and a unique pattern of monoalleic T cell γ gene rearrangement was observed. The rearrangement involves the recently described Vγ5 segment and the Jγ1 joining segment, which is located upstream of the Cγ1 constant region. To determine the molecular structure present on DS6, an immunoprecipitation was performed with monoclonal anti-CD3 antibody and a rabbit antiserum raised against γ protein. We have observed, in association with the CD3 complex, a 90 kDa structure which under reducing conditions resolves into three subunits of 45, 40 and 37 kDa. We demonstrated that the rabbit anti-γ serum only immunoprecipitates the two lower bands. The upper band corresponds to a presently undefined T cell receptor chain. Next, we showed the non major histocompatibility complex (MHC)-restricted cytolytic activity exhibited by these CTJ3+ CD4- CD8- cloned T cells and inhibition of the natural killer (NK)-like activity by the anti-CD3 monoclonal antibody. The triggering of CD2 or CD3 molecules increased IL-2 receptor expression on DS6 but failed to induce cell proliferation. This contrasts with recent results obtained with γ-expressing T cell clones and illustrates the functional heterogeneity of the cells bearing the second T cell receptor.
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