title: Characterization of an anti-rainbow trout (Oncorhynchus mykiss) CD3ε monoclonal antibody.
abstract: This study characterizes a monoclonal antibody (mAb) produced against the cytoplasmic tail region of the epsilon chain of the CD3 (CD3ε) transmembrane protein found on T lymphocytes of rainbow trout (Oncorhynchus mykiss). Flow cytometry and fluorescent microscopy conducted on trout leukocytes with the anti-trout CD3ε mAb showed a distinctive population of IgM(-) CD3e(+) lymphocytes fitting the expected profile of T-cells. Immunoprecipitation of lysates derived from trout lymphocytes revealed a 19 kDa protein and peptide analysis confirmed its specificity for CD3ɛ. In vitro proliferation assays with T-cell mitogens, ConA and PHA, resulted in a 3 fold increase in the percentage of CD3ɛ+ lymphocytes compared to LPS and control cultures. The mAb characterized in this study will be useful in further elucidation for both the role and distribution of T lymphocytes in the teleost immune system.
authors.affiliation: Department of Biology, University of Massachusetts Dartmouth, 285 Old Westport Road, Dartmouth, MA 02747, USA.
authors.forename: T
authors.initials: T
authors.lastname: Boardman
authors.forename: C
authors.initials: C
authors.lastname: Warner
authors.forename: F
authors.initials: F
authors.lastname: Ramirez-Gomez
authors.forename: J
authors.initials: J
authors.lastname: Matrisciano
authors.forename: E
authors.initials: E
authors.lastname: Bromage
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D005434:Flow Cytometry / Q000662:veterinary; D047468:Immunoprecipitation / Q000662:veterinary; D008213:Lymphocyte Activation / Q000187:drug effects; D018942:Macrolides / Q000494:pharmacology; D008856:Microscopy, Fluorescence / Q000662:veterinary; D017686:Oncorhynchus mykiss / Q000276:immunology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D018942:Macrolides; D010835:Phytohemagglutinins; C036978:concanamycin A