Definition of the HLA-A2 restricted peptides recognized by human CD8+ effector T cells by flow-assisted sorting of the CD8+ CD45RA+ CD28- T cell subpopulation.
In response to antigenic stimulation, naive MHC-class I restricted and antigen-specific CD8+ CD45RA+ CD28+ T cells undergo clonal expansion, differentiate into CD8+ CD45RO+ memory T cells and convert to CD8+ CD45RA+ CD28- T cells displaying potent immune effector functions upon re-encounter with the nominal antigen. We show that the effector CD8+ CD45RA+ CD28- T cell subset is expanded in peripheral blood lymphocytes (PBL) from patients with human papilloma virus (HPV)+ cervical lesions as well as in PBL from patients with pulmonary tuberculosis. Flow-cytometric cell sorted CD8+ CD45RA+ CD28- and CD8+ CD45RA+ CD28- T cells were tested for recognition of HLA-A2 restricted peptides derived either from the human papillomavirus (HPV)16-E7 gene product, or from M. tuberculosis antigens. Mostly CD8+ CD45+ CD28- T cells define antigen/peptide-specific and MHC-restricted responses. These data were confirmed in PBL from patients with tuberculosis using HLA-A2 tetramer-complexes loaded with a peptide from the M. tuberculosis Ag85b antigen by flow cytometry. The sorting of this T cell subset enables to determine the fine specificity of CD8+ effector T cells without the need for in vitro manipulation.
Höhn
H
H
Department of Medical Microbiology, University of Mainz, Germany.
Jülch
M
M
Pilch
H
H
Kortsik
C
C
Tully
G
G
Neukirch
C
C
Freitag
K
K
Maeurer
M
M
D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology*; D018106:CD28 Antigens; D002454:Cell Differentiation; D002455:Cell Division; D005260:Female; D005434:Flow Cytometry; D015789:HLA-A2 Antigen; D006801:Humans; D016130:Immunophenotyping; D017493:Leukocyte Common Antigens; D009169:Mycobacterium tuberculosis / Q000276:immunology; D030361:Papillomavirus Infections / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D014397:Tuberculosis, Pulmonary / Q000276:immunology; D002583:Uterine Cervical Neoplasms / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018106:CD28 Antigens; D015789:HLA-A2 Antigen; D014764:Viral Proteins; D017493:Leukocyte Common Antigens