B cell and immunoglobulin heterogeneity in carp ( Cyprinus carpio L); An immuno(cyto)chemical study

• Published in: Developmental and comparative immunology 1995, Vol.19 (1), p.97-108
• Main Authors: Koumans-van Diepen, JoséC.E., Egberts, Egbert, Peixoto, Bernardo R., Taverne, Nico, Rombout, Jan H.W.M.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibody Diversity; Antibody Specificity; B cells; B-Lymphocyte Subsets - immunology; Blotting, Western; Carps - growth & development; Carps - immunology; Chromatography, Affinity; Fish; Flow Cytometry; Gene Rearrangement, B-Lymphocyte; Hemocyanins - immunology; Heterogeneity; Immunocytochemistry; Immunoglobulin; Immunoglobulin Heavy Chains - immunology; Immunohistochemistry; Lymphoid Tissue - cytology; Lymphoid Tissue - growth & development; Lymphoid Tissue - immunology; Microscopy, Fluorescence; Ontogeny; Plasma Cells - immunology; Spleen - cytology; Spleen - growth & development; Spleen - immunology
• ISSN: 0145-305X; 1879-0089
• DOI: 10.1016/0145-305X(94)00061-J

B cell and immunoglobulin (Ig) heterogeneity was demonstrated in carp, Cyprinus carpio L., using two monoclonal antibodies (MAbs; WC14, WCI12) produced against carp serum Ig. Immunochemical results showed that both WCI4 and WCI12 react with a protein determinant on the heavy chain of Ig (relative molecular mass ~70,000). Immunofluorescence microscopic and flow cytometric analyses of lymphoid cells suggest three distinct subpopulations of B cells and plasma cells: WCI4 +12 − cells, WCI4 −12 + cells, and WCI4 +12 + cells. WCI4 −12 + and WCI4 +12 + anti-DNP antibody-secreting cells were also demonstrated with the ELISPOT assay in pronephros and spleen cell suspensions from primary immunised carp. Affinity chromatography of carp serum and sequential immunoprecipitation of 125I-labelled peripheral blood leucocyte (PBL) membrane proteins only indicated the presence of two antigenically different Ig molecules i.e., WCI4 −12 + and WCI4 +2 + molecules. WCI4 +12- molecules could not be detected by affinity chromatography or immunoprecipitation. During ontogeny, a shift in percentages of WCI4 +12 − and WCI4 −12 + cells was found in the spleen and the pronephros. In these organs, WCI4 +12 − cells formed the majority of B cells at 2 weeks of age, but the percentages of this cell type decreased during ontogeny. On the other hand, the percentages of WCI4 −12 + cells increased during development, and these cells became the major population of B cells from 13 weeks Address correspondence to Dr. J.H.W.M. Rombout. onward. The proportion of WCI4 +12 + cells remained almost constant during ontogeny. The distribution of B cell subpopulations in blood was more or less stable at all ages. The functional significance of Ig heterogeneity in fish and in particular carp is discussed.