title: MHC class II tetramers containing influenza hemagglutinin and EBV EBNA1 epitopes detect reliably specific CD4(+) T cells in healthy volunteers.
abstract: Tracking antigen specific T cells with major histocompatibility complex (MHC) tetramers has provided us with insights into the dynamics of the adaptive immune system and holds great promise to aid in patient management and drug and vaccine development. Progress has been made primarily using MHC class I tetramers to monitor CD8(+) T cells, whereas corresponding efforts to stain CD4(+) T cells with class II tetramers have not been as successful. Two major reasons have been proposed for this lack of progress: (1). The frequency of antigen-specific CD4(+) T cells is lower than the frequency of CD8(+) T cells and (2). some, but not all, antigen- specific CD4(+) T cells can bind tetramer because of low functional avidity. In this study, we asked if CD4(+) T cells specific for common human viruses (e.g., influenza and Epstein-Barr) can be detected in healthy individuals previously exposed to them. We were able to clearly detect specific CD4(+) T cells in all donors after in vitro expansion of peripheral blood mononuclear cells. Furthermore, we observe a clear separation of tetramer negative and tetramer positive CD4(+) T cells in most samples similar to patterns commonly seen with class I tetramers. The data indicate that MHC class II tetramers can be used reliably for the identification of CD4(+) T cells specific for ubiquitous infectious agents in normal donors.
authors.affiliation: Cell Analysis Development Center, Beckman Coulter, Inc, San Diego, CA 92121, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Ye
authors.forename: Suha
authors.initials: S
authors.lastname: Kasey
authors.forename: Sumit
authors.initials: S
authors.lastname: Khurana
authors.forename: Nga T
authors.initials: NT
authors.lastname: Nguyen
authors.forename: Scott
authors.initials: S
authors.lastname: Schubert
authors.forename: C Thomas
authors.initials: CT
authors.lastname: Nugent
authors.forename: Kristine
authors.initials: K
authors.lastname: Kuus-Reichel
authors.forename: Johannes
authors.initials: J
authors.lastname: Hampl
mesh_terms: D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000737:chemistry; D006684:HLA-DR Antigens / Q000737:chemistry / Q000276:immunology; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D006389:Hemagglutinins, Viral / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000737:chemistry / Q000187:drug effects / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D013388:Succinimides / Q000737:chemistry
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D019309:Epstein-Barr Virus Nuclear Antigens; D005452:Fluoresceins; D006684:HLA-DR Antigens; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D000949:Histocompatibility Antigens Class II; D007376:Interleukin-2; D010446:Peptide Fragments; D013388:Succinimides; C086595:influenza hemagglutinin (306-318); C417029:EBV-encoded nuclear antigen 1