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

Dev Comp Immunol. 1995 Jan-Feb;19(1):97-108. doi: 10.1016/0145-305x(94)00061-j.

Abstract

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 approximately 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+12+ 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 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.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibody Diversity*
  • Antibody Specificity
  • B-Lymphocyte Subsets / immunology*
  • Blotting, Western
  • Carps / growth & development
  • Carps / immunology*
  • Chromatography, Affinity
  • Flow Cytometry
  • Gene Rearrangement, B-Lymphocyte
  • Hemocyanins / immunology
  • Immunoglobulin Heavy Chains / immunology
  • Immunohistochemistry
  • Lymphoid Tissue / cytology
  • Lymphoid Tissue / growth & development
  • Lymphoid Tissue / immunology
  • Microscopy, Fluorescence
  • Plasma Cells / immunology
  • Spleen / cytology
  • Spleen / growth & development
  • Spleen / immunology

Substances

  • 2,4-dinitrophenyl keyhole limpet hemocyanin
  • Antibodies, Monoclonal
  • Immunoglobulin Heavy Chains
  • Hemocyanins