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2V
2 TCR1
* Department of Microbiology and Immunology, Center for Primate Biomedical Research, University of Illinois College of Medicine Chicago, Chicago, Illinois 60612; and
Research Resources Center, University of Illinois College of Medicine Chicago, Chicago, Illinois 60612
Although microbial (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) can activate primate V
2V
2 T cells, molecular mechanisms by which HMBPP interacts with V2V2 T cells remain poorly characterized. Here, we developed soluble, tetrameric V2V2 TCR of rhesus macaques to define HMBPP/APC interaction with V2V2 TCR. While exogenous HMBPP was associated with APC membrane in an appreciable affinity, the membrane-associated HMBPP readily bound to the V2V2 TCR tetramer. The V2V2 TCR tetramer was shown to bind stably to HMBPP presented on membrane by various APC cell lines from humans and nonhuman primates but not those from mouse, rat, or pig. The V2V2 TCR tetramer also bound to the membrane-associated HMBPP on primary monocytes, B cells and T cells. Consistently, endogenous phosphoantigen produced in Mycobacterium-infected dendritic cells was transported and presented on membrane, and bound stably to the V2V2 TCR tetramer. The capability of APC to present HMBPP for recognition by V2V2 TCR was diminished after protease treatment of APC. Thus, our studies elucidated an affinity HMBPP-APC association conferring stable binding to the V2V2 TCR tetramer and the protease-sensitive nature of phosphoantigen presentation. The findings defined APC presentation of phosphoantigen HMBPP to V2V2 TCR.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This work was supported by National Institutes of Health RO1 Grants HL64560 and RR13601 (both to Z.W.C.).
2 Current address: Department of Microbiology, Sun Yat-sen University, P.R. China.
3 Address correspondence and reprint requests to Prof. Z. W. Chen, Department of Microbiology and Immunology, Center for Primate Biomedical Research, University of Illinois College of Medicine Chicago, Chicago, Illinois 60612. E-mail address: zchen{at}uic.edu
4 Abbreviations used in this paper: HMBPP, (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate; IPP, isopentenyl pyrophosphate; DC, dendritic cell; BCG, bacillus Calmette-Guérin.
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