Characterization of an anti-rainbow trout (Oncorhynchus mykiss) CD3ε monoclonal antibody

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.