Microsystem for in vitro primary and secondary immunization of channel catfish (Ictalurus punctatus) leukocytes with hapten-carrier conjugates.
Methods are described for the in vitro generation and detection of antibody-secreting cells (PFC) from channel catfish. Hapten-specific PFC can readily be enumerated by an indirect plaque assay employing rabbit antiserum to catfish Ig and guinea pig complement. A modified Mishell-Dutton-type culture system was developed for effectively generating significant in vitro anti-hapten PFC responses with catfish leukocytes at 27 degrees C. The classical hapten-carrier effect and primary responses to both TI and TD antigens were demonstrable with catfish cells. Variables found to be important with catfish cells included the serum supplement, cell densities and, to a lesser extent, antigen form. Optimistically these methods will prove useful in attempts to delineate the functional roles of different lymphocyte subpopulations in fish.
D000818:Animals; D000921:Antibody-Producing Cells / Q000276:immunology; D002352:Carrier Proteins / Q000008:administration & dosage / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes; D005260:Female; D005399:Fishes / Q000276:immunology*; D006241:Haptens / Q000008:administration & dosage / Q000276:immunology*; D006462:Hemolytic Plaque Technique; D007117:Immunization, Secondary / Q000379:methods; D007700:Kinetics; D007958:Leukocyte Count; D007962:Leukocytes / Q000276:immunology*; D008297:Male
D002352:Carrier Proteins; D000939:Epitopes; D006241:Haptens