Lymphoblastoid cell supernatants increase expression of C3b receptors on human polymorphonuclear leucocytes: direct binding studies with 125I-C3b.
Human PMN incubated in culture supernatants of the Raji long-term human lymphoblastoid cell line showed increased rosette formation with sheep erythrocytes coated with C3b (EIgM C4b3b) but no change in rosette formation with IgG-coated erythrocytes. This suggested a specific increase in cell surface C3b receptors, which was further investigated using 125I-C3b for direct binding studies. The results confirmed that specific binding of 125I-C3b to PMN incubated in culture supernatants increased up to three- to four-fold over binding to PMN incubated in control media alone. Scatchard analysis revealed that the apparent Ka for supernatant-treated cells, 3.36 +/- 0.89 X 10(7) L/M did not differ from the Ka for cells incubated in control media, 3.76 +/- 0.75 X 10(7) L/M, suggesting an increase in a single class of C3b receptors. Kinetic studies revealed that the active factor was present within 24 hr of culture of the Raji cells, and that neutrophils incubated in culture supernatants increased their C3b receptors continuously for up to 4 hr, the longest interval tested. The effect of the culture supernatant was lost with dilution beyond eight- to 10-fold. The results suggest that culture supernatants of this long-term lymphoblastoid cell line contain soluble factors that induce increased expression of C3b receptors on PMN and may thus serve as a model for study of important physiologic effects of lymphocyte products on PMN in vivo.
D002460:Cell Line; D003470:Culture Media; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007457:Iodine Radioisotopes; D007700:Kinetics; D009504:Neutrophils / Q000276:immunology*; D011951:Receptors, Complement / Q000276:immunology*; D017463:Receptors, Complement 3b; D011971:Receptors, Immunologic / Q000276:immunology; D012397:Rosette Formation
D003470:Culture Media; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D011971:Receptors, Immunologic