A MOUSE MONOCLONAL-ANTIBODY SPECIFIC FOR AN ALLOTYPIC DETERMINANT OF THE IGHA ALLELE OF MURINE IGM - GENETIC AND FUNCTIONAL-ANALYSIS OF IGH-6A EPITOPES USING ANTI-IGM MONOCLONAL-ANTIBODIES

An IgG1 mouse monoclonal antibody (MAb) specific for a mouse IgM allotypic determinant in the a, c, f, g, h, and j haplotypes was derived from a fusion of SP2/0-Ag14 mouse myeloma cells with C57BL/6 mouse spleen cells (Igh-C(b)) immune to TC31, a MAb of the IgM(a) allotype. MAb from one hybridoma derived from this fusion (designated DS1) was demonstrated to bind in an ELISA to immunoglobulin bearing the IgM(a) allotype (TC31, MOPC104E), but not to immunoglobulin bearing the IgM(b) allotype (C. BPC112). Fluorescein-conjugated DS1 was shown to bind to the surface of BALB/cByJ splenic B cells, but was shown to have negligible binding on C57BL/6J cells. Similarily, DS1-conjugated Sepharose beads were able to stimulate in vitro proliferation of BALB/c, but not C57BL/6 splenic B cells. DS1 was unable to bind to spleen cells from BALB/c allotype congenic strains, BAB/14 (Igh-C(b)) and C.AL-20 (Igh-C-degrees), demonstrating that DS1 recognizes a determinant under the control of a gene linked to the Igh-C gene complex. Using sera from recombinant inbred lines, the determinant defined by DS1 was shown to be linked to the Igh-1 locus. Furthermore, the determinant was localized to the CH1 domain of the mu heavy chain. Sera from BALB/cByJ, NMRI, CBA/J, SEA/GnJ, RIIIs/J, and CE/J mouse strains were shown to bind to DS1 in an ELISA, while sera from A/J, SJL/J, NZB/B1NJ, AKR/J, C57BL/6J, and C57BL/10SnJ mouse strains did not bind to DS1. From these data we propose that DS1 is reactive with specificity Igh-6.1, which was originally defined by an allotypic antiserum developed by Black et al. (Immunogenetics 7:213, 1978).