title: First Demonstration of Antigen Induced Cytokine Expression by CD4-1+ Lymphocytes in a Poikilotherm: Studies in Zebrafish (Danio rerio).
abstract: Adaptive immunity in homeotherms depends greatly on CD4+ Th cells which release cytokines in response to specific antigen stimulation. Whilst bony fish and poikilothermic tetrapods possess cells that express TcR and CD4-related genes (that exist in two forms in teleost fish; termed CD4-1 and CD4-2), to date there is no unequivocal demonstration that cells equivalent to Th exist. Thus, in this study we determined whether CD4-1+ lymphocytes can express cytokines typical of Th cells following antigen specific stimulation, using the zebrafish (Danio rerio). Initially, we analyzed the CD4 locus in zebrafish and found three CD4 homologues, a CD4-1 molecule and two CD4-2 molecules. The zfCD4-1 and zfCD4-2 transcripts were detected in immune organs and were most highly expressed in lymphocytes. A polyclonal antibody to zfCD4-1 was developed and used with an antibody to ZAP70 and revealed double positive cells by immunohistochemistry, and in the Mycobacterium marinum disease model CD4-1+ cells were apparent surrounding the granulomas typical of the infection. Next a prime-boost experiment, using human gamma globulin as antigen, was performed and revealed for the first time in fish that zfCD4-1+ lymphocytes increase the expression of cytokines and master transcription factors relevant to Th1/Th2-type responses as a consequence of boosting with specific antigen.
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Sohye
authors.initials: S
authors.lastname: Yoon
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Suman
authors.initials: S
authors.lastname: Mitra
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Cathy
authors.initials: C
authors.lastname: Wyse
authors.affiliation: Vertebrate Antibodies Ltd, Zoology Building, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Ayham
authors.initials: A
authors.lastname: Alnabulsi
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.affiliation: VU University, Medical Center, Department of Medical Microbiology & Infection Control, 1081 BT Amsterdam, Amsterdam, The Netherlands.
authors.forename: Eveline M
authors.initials: EM
authors.lastname: Weerdenburg
authors.affiliation: VU University, Medical Center, Department of Medical Microbiology & Infection Control, 1081 BT Amsterdam, Amsterdam, The Netherlands.
authors.forename: Astrid M
authors.initials: AM
authors.lastname: van der Sar
authors.affiliation: Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, Washington, United States of America.
authors.forename: Difei
authors.initials: D
authors.lastname: Wang
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Secombes
authors.affiliation: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, Aberdeen, United Kingdom.
authors.forename: Steve
authors.initials: S
authors.lastname: Bird
mesh_terms: D056704:Adaptive Immunity; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D015496:CD4-Positive T-Lymphocytes / Q000145:classification / Q000276:immunology* / Q000382:microbiology / Q000473:pathology; D016207:Cytokines / Q000096:biosynthesis* / Q000276:immunology; D056426:Genetic Loci / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009165:Mycobacterium Infections, Nontuberculous / Q000235:genetics / Q000276:immunology* / Q000382:microbiology / Q000662:veterinary*; D019910:Mycobacterium marinum / Q000276:immunology; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000276:immunology*; D016415:Sequence Alignment; D058408:Th1-Th2 Balance; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000235:genetics / Q000276:immunology; D015027:Zebrafish / Q000145:classification / Q000235:genetics / Q000276:immunology; D029961:Zebrafish Proteins / Q000235:genetics / Q000276:immunology; D005719:gamma-Globulins / Q000008:administration & dosage
chemical_list: D000906:Antibodies; D016207:Cytokines; D000949:Histocompatibility Antigens Class II; D012333:RNA, Messenger; D029961:Zebrafish Proteins; D005719:gamma-Globulins; D051746:ZAP-70 Protein-Tyrosine Kinase