Antibody synthesis in roach ( Rutilus rutilus); analysis of antibody secreting cells in lymphoid organs with ELISPOT-assay

Abstract

The roach ( Rutilus rutilus L.) which is a cyprinid fish, was immunised with bovine γ-globulin (BGG) and the antibody synthesis was studied by counting the number of specific antibody secreting cells (SASC) in the spleen and anterior kidney, and by measuring the antibody concentration in the circulation. SASCs and the total number of immunoglobulin secreting cells (TISC) were counted with the ELISPOT (enzyme-linked immunospot) assay, and anti-BGG antibodies and the concentration of immunoglobulin in sera were assayed by ELISA (enzyme-linked immunosorbent assay). The present modification of the ELISPOT-assay takes advantage of biotin-avidin amplification and yields easily detectable and nonfading spots that may be counted even after several weeks of storage. It is also possible to count the number of SASC and TISC in different organs. In this study, immunisation of the roach with protein antigen via the intraperitoneal route resulted in a similar response in the spleen and in the kidney. There was no preferential organ distribution of the number of SASC or TISC and no difference in the timing of the response in the organs. The humoral response due to immunisation in the serum correlated with cellular responses in the organs.