caa a22 4500 467906874 CHVBK 20180406152836.0 cr unu---uuuuu 170328e20060901xx s 000 0 eng 10.1007/s00251-006-0140-3 doi (NATIONALLICENCE)springer-10.1007/s00251-006-0140-3 Quantitation of Anaplasma marginale major surface protein (MSP)1a and MSP2 epitope-specific CD4+ T lymphocytes using bovine DRB3*1101 and DRB3*1201 tetramers [Elektronische Daten] [Junzo Norimine, Sushan Han, Wendy Brown] Antigen-specific CD4+ T cells play a critical role in protective immunity to many infectious pathogens. Although the antigen-specific CD4+ T cells can be measured by functional assays such as proliferation or cytokine enzyme-linked immunospot, such assays are limited to a specific function and cannot quantify anergic or suppressed T cells. In contrast, major histocompatiblity complex (MHC) class II tetramers can enumerate epitope-specific CD4+ T cells independent of function. In this paper, we report the construction of bovine leukocyte antigen MHC class II tetramers using a novel mammalian cell system to express soluble class II DRA/DRB3 molecules and defined immunodominant peptide epitopes of Anaplasma marginale major surface proteins (MSPs). Phycoerythrin-labeled tetramers were either loaded with exogenous peptide or constructed with the peptide epitope linked to the N terminus of the DRB3 chain. A DRB3*1101 tetramer loaded with MSP1a peptide F2-5B (ARSVLETLAGHVDALG) and DRB3*1201 tetramers loaded with MSP1a peptide F2-1-1b (GEGYATYLAQAFA) or MSP2 peptide P16-7 (NFAYFGGELGVRFAF) specifically stained antigen-specific CD4+ T cell lines and clones. Tetramers constructed with the T-cell epitope linked to the DRB3 chain were slightly better at labeling CD4+ T cells. In one cell line, the number of tetramer-positive T cells increased to approximately 94% of the CD4+ T cells after culture for 21weeks with specific antigen. This novel technology should be useful to track the fate of antigen-specific CD4+ T-cell responses in cattle after immunization or infection with persistent pathogens, such as A. marginale, that modulate the host immune response. Springer-Verlag, 2006 DRB3 nationallicence MHC class II nationallicence Tetramer nationallicence Anaplasma marginale nationallicence Cattle nationallicence Norimine Junzo Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040, Pullman, WA, USA aut Han Sushan Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040, Pullman, WA, USA aut Brown Wendy Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040, Pullman, WA, USA aut Immunogenetics Springer-Verlag 58/9(2006-09-01), 726-739 0093-7711 58:9<726 2006 58 251 https://doi.org/10.1007/s00251-006-0140-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00251-006-0140-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Norimine Junzo Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040, Pullman, WA, USA aut NATIONALLICENCE 700 1- Han Sushan Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040, Pullman, WA, USA aut NATIONALLICENCE 700 1- Brown Wendy Department of Veterinary Microbiology and Pathology, Washington State University, 99164-7040, Pullman, WA, USA aut NATIONALLICENCE 773 0- Immunogenetics Springer-Verlag 58/9(2006-09-01), 726-739 0093-7711 58:9<726 2006 58 251 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer