title: In situ detection of autoreactive CD4 T cells in brain and heart using major histocompatibility complex class II dextramers.
abstract: This report demonstrates the use of major histocompatibility complex (MHC) class II dextramers for detection of autoreactive CD4 T cells in situ in myelin proteolipid protein (PLP) 139-151-induced experimental autoimmune encephalomyelitis (EAE) in SJL mice and cardiac myosin heavy chain-α (Myhc) 334-352-induced experimental autoimmune myocarditis (EAM) in A/J mice. Two sets of cocktails of dextramer reagents were used, where dextramers(+) cells were analyzed by laser scanning confocal microscope (LSCM): EAE, IA(s)/PLP 139-151 dextramers (specific)/anti-CD4 and IA(s)/Theiler's murine encephalomyelitis virus (TMEV) 70-86 dextramers (control)/anti-CD4; and EAM, IA(k)/Myhc 334-352 dextramers/anti-CD4 and IA(k)/bovine ribonuclease (RNase) 43-56 dextramers (control)/anti-CD4. LSCM analysis of brain sections obtained from EAE mice showed the presence of cells positive for CD4 and PLP 139-151 dextramers, but not TMEV 70-86 dextramers suggesting that the staining obtained with PLP 139-151 dextramers was specific. Likewise, heart sections prepared from EAM mice also revealed the presence of Myhc 334-352, but not RNase 43-56-dextramer(+) cells as expected. Further, a comprehensive method has also been devised to quantitatively analyze the frequencies of antigen-specific CD4 T cells in the 'Z' serial images.
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska, Lincoln.
authors.forename: Chandirasegaran
authors.initials: C
authors.lastname: Massilamany
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska, Lincoln.
authors.forename: Arunakumar
authors.initials: A
authors.lastname: Gangaplara
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska, Lincoln.
authors.forename: Ting
authors.initials: T
authors.lastname: Jia
authors.affiliation: Center for Biotechnology, University of Nebraska, Lincoln.
authors.forename: Christian
authors.initials: C
authors.lastname: Elowsky
authors.affiliation: Nebraska Center for Virology and School of Biological Sciences, University of Nebraska, Lincoln.
authors.forename: Qingsheng
authors.initials: Q
authors.lastname: Li
authors.affiliation: Center for Biotechnology, University of Nebraska, Lincoln.
authors.forename: You
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: School of Veterinary Medicine and Biomedical Sciences, University of Nebraska, Lincoln; jayreddy@unl.edu.
authors.forename: Jay
authors.initials: J
authors.lastname: Reddy
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology / Q000473:pathology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008805:Mice, Inbred A; D018991:Myelin Proteolipid Protein; D009206:Myocardium / Q000166:cytology / Q000276:immunology*; D010446:Peptide Fragments
chemical_list: D018984:Epitopes, T-Lymphocyte; D000949:Histocompatibility Antigens Class II; D018991:Myelin Proteolipid Protein; D010446:Peptide Fragments; C435023:myelin proteolipid protein (139-151)