IgM expressed by leukemic CD5(+) B cells binds mouse immunoglobulin light chain.
Mouse immunoglobulin (Ig) molecules have previously been shown to bind to the surface of CD5(+) B cells from patients with B-cell chronic lymphocytic leukemia (B-CLL). The results indicated that surface IgM was involved in the interaction and suggested the phenomenon was an example of the polyreactive binding capacity of the surface Ig (sIg) expressed by these malignant cells. This article describes the further characterization of the interaction between human IgM and mouse Ig molecules and subunits. Mouse Ig molecules of both kappa and lambda light chain classes bound to the B-CLL cell surface. The dissociation constant for the interaction of mouse IgG1 (K121) with the B-CLL cell surface was 3.6 x 10(-7) M. To confirm the involvement of the human IgM expressed by the B-CLL cells in the interaction, the malignant cells were stimulated in vitro to induce secretion of human IgM. Enzyme immunoassay was used to show that secreted human IgM bound to intact mouse Ig, as occurred with the cell surface analysis. The mouse Ig epitope recognized by the purified secreted human IgM was shown by Western blot analysis to be located on the light chain of the mouse Ig molecule and to be conformationally dependent. K121 light chain was cloned and expressed in E. coli and the recombinant light chain bound to the surface of CLL B cells. The results confirm that human IgM is the reactive ligand in the interaction with mouse Ig and indicate that the interaction of polyreactive IgM with mouse IgG occurs via the light chain component of IgG.
Weston
K M
KM
Immunobiology Unit, Department of Cell and Molecular Biology University of Technology, Sydney, Westbourne Street, Gore Hill, 2065, NSW, Australia.
Tangye
S G
SG
Dunn
R D
RD
Smith
A
A
Morris
M B
MB
Raison
R L
RL
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D018956:CD5 Antigens / Q000276:immunology; D003001:Cloning, Molecular; D000939:Epitopes; D004926:Escherichia coli; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D007075:Immunoglobulin M / Q000276:immunology* / Q000378:metabolism; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry / Q000276:immunology; D007146:Immunoglobulin lambda-Chains / Q000737:chemistry / Q000276:immunology; D007148:Immunoglobulin mu-Chains / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology*; D008024:Ligands; D051379:Mice; D011487:Protein Conformation; D014407:Tumor Cells, Cultured
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D018956:CD5 Antigens; D000939:Epitopes; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D007148:Immunoglobulin mu-Chains; D008024:Ligands; D016650:Fluorescein-5-isothiocyanate