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oai:docta.ucm.es:20.500.14352/1074322024-08-07T23:54:14Zcom_20.500.14352_14col_20.500.14352_15

Biochemical differences in the αβ T cell receptor-CD3 surface complex between CD8+ and CD4+ human mature T lymphocytes Zapata, David A. Schamel, Wolfgang W.A. Torres, Pilar S. Alarcón, Balbino Rossi, Nineth E. Navarro, María N. Toribio, María L. Regueiro González-Barros, José Ramón 616-097 Inmunología 2412 Inmunología We have reported the existence of biochemical and conformational differences in the alphabeta T cell receptor (TCR) complex between CD4(+) and CD8(+) CD3gamma-deficient (gamma(-)) mature T cells. In the present study, we have furthered our understanding and extended the observations to primary T lymphocytes from normal (gamma(+)) individuals. Surface TCR.CD3 components from CD4(+) gamma(-) T cells, other than CD3gamma, were detectable and similar in size to CD4(+) gamma(+) controls. Their native TCR.CD3 complex was also similar to CD4(+) gamma(+) controls, except for an alphabeta(deltaepsilon)(2)zeta(2) instead of an alphabetagammaepsilondeltaepsilonzeta(2) stoichiometry. In contrast, the surface TCRalpha, TCRbeta, and CD3delta chains of CD8(+) gamma(-) T cells did not possess their usual sizes. Using confocal immunofluorescence, TCRalpha was hardly detectable in CD8(+) gamma(-) T cells. Blue native gels (BN-PAGE) demonstrated the existence of a heterogeneous population of TCR.CD3 in these cells. Using primary peripheral blood T lymphocytes from normal (gamma(+)) donors, we performed a broad epitopic scan. In contrast to all other TCR.CD3-specific monoclonal antibodies, RW2-8C8 stained CD8(+) better than it did CD4(+) T cells, and the difference was dependent on glycosylation of the TCR.CD3 complex but independent of T cell activation or differentiation. RW2-8C8 staining of CD8(+) T cells was shown to be more dependent on lipid raft integrity than that of CD4(+) T cells. Finally, immunoprecipitation studies on purified primary CD4(+) and CD8(+) T cells revealed the existence of TCR glycosylation differences between the two. Collectively, these results are consistent with the existence of conformational or topological lineage-specific differences in the TCR.CD3 from CD4(+) and CD8(+) wild type T cells. The differences may be relevant for cis interactions during antigen recognition and signal transduction. Depto. de Inmunología, Oftalmología y ORL Fac. de Medicina TRUE pub 2024-08-07T11:31:00Z 2024-08-07T11:31:00Z 2004-02-27 journal article VoR https://hdl.handle.net/20.500.14352/107432 XXXX-XXXX 10.1074/jbc.M311455200 eng Zapata DA, Schamel WW, Torres PS, Alarcón B, Rossi NE, Navarro MN, Toribio ML, Regueiro JR. Biochemical differences in the alphabeta T cell receptor.CD3 surface complex between CD8+ and CD4+ human mature T lymphocytes. J Biol Chem. 2004 Jun 4;279(23):24485-92. doi: 10.1074/jbc.M311455200. Epub 2004 Apr 1. PMID: 15060077. Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ open access application/pdf Elsevier