Characterization of the superoxide-generating system in human peripheral lymphocytes and lymphoid cell lines.
In B lymphocytes, but not T lymphocytes, isolated from human peripheral blood, we detected the four protein components essential for "the respiratory burst" by immunoblot analyses using peptide-directed antibodies. These are two membrane proteins, namely, 91- and 22-kDa subunits of cytochrome b558, and two cytosolic proteins with molecular masses of 47 and 65 kDa. Like in neutrophils, cytochrome b558 was expressed on the cell surface of peripheral B lymphocytes. Mean amounts (n = 8) of the 91-, 22-, 47-, and 65-kDa proteins, respectively, in peripheral B lymphocytes calculated from intensity of the blots were 0.011 +/- 0.003, 0.026 +/- 0.006, 0.179 +/- 0.022, and 0.039 +/- 0.013 relative to those in neutrophils on the basis of cell number. Epstein-Barr virus (EBV)-transformed cell lines derived from normal B lymphocytes and some B cell lines also possessed cytochrome b558 and two cytosolic proteins. Isolated human peripheral B lymphocytes generated the superoxide anion upon cross-linking of surface antigens such as IgM, IgD, IgG, HLA-DR, and CD19. EBV-transformants derived from normal peripheral B lymphocytes and B lymphoid cell lines also generated the superoxide anion when stimulated with various antibodies against surface antigens. These results indicate that peripheral B lymphocytes have substantial amounts of a superoxide-generating system identical to that in phagocytes and that the system is stimulated to generate the superoxide anion by the cross-linking of clonally expressed surface immunoglobulins or of certain surface antigens.
Kobayashi
S
S
Institute of Medical Science, University of Tokyo.
Imajoh-Ohmi
S
S
Kuribayashi
F
F
Nunoi
H
H
Nakamura
M
M
Kanegasaki
S
S
D000954:Antigens, Surface / Q000378:metabolism; D001402:B-Lymphocytes / Q000378:metabolism* / Q000821:virology; D001798:Blood Proteins / Q000032:analysis / Q000378:metabolism; D002460:Cell Line; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D003573:Cytochrome b Group / Q000032:analysis / Q000097:blood; D003600:Cytosol / Q000737:chemistry; D005434:Flow Cytometry; D004854:Herpesvirus 4, Human; D006801:Humans; D015151:Immunoblotting; D016399:Lymphoma, T-Cell / Q000378:metabolism; D046911:Macromolecular Substances; D019255:NADPH Oxidases; D009504:Neutrophils / Q000378:metabolism; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D001798:Blood Proteins; D003573:Cytochrome b Group; D046911:Macromolecular Substances; D013481:Superoxides; C036888:cytochrome b558; D019255:NADPH Oxidases