Generation of monoclonal antibodies to coelomocytes of the purple sea urchin Arbacia punctulata: Characterization and phenotyping

Abstract

Cellular cytotoxicity is a key component of animal innate immune responses that is one of the first lines of defense against invaders. There is increasing interest in the study of the cellular immune response, particularly non-specific cytotoxic cells and natural killer cells and their receptors. Studies of non-specific cytotoxic cell and natural killer cell recognition and killing (and the receptors involved) will reveal new and important insights into cellular mechanisms of host defense. Here we describe mAbs specific for coelomocyte sub-populations of the purple sea urchin, Arbacia punctulata, using highly purified coelomocyte populations as the antigen source. Monoclonal antibodies were selected using flow cytometric screening methods. Several of the mAbs were shown to bind to two sub-types of coelomocytes when assayed by fluorescence microscopy. Furthermore, these mAbs inhibited coelomocyte cytotoxicity against vertebrate target cells in a functional assay. The mAbs have been used in immunoprecipitation studies.