A cell-ELISA for the quantification of adherent murine macrophages and the surface expression of antigens.
The present study was performed in order to establish whether a cell-ELISA could be used to determine the expression of antigens by adherent murine peritoneal macrophages and also quantify the numbers of such macrophages. Accurate determination of the number of adherent macrophages proved to be possible with a cell-ELISA designed to assess complement receptor type III (CRIII) expression. Expression of CRIII was considerably more sensitive than determination of the cell-protein or DNA content as a measure of the number of adherent macrophages. For the calculation of the expression of CRIII, Ia antigen, and antigen F4/80 by resident and activated macrophages, use was made of the linear part of the curve obtained when the numbers of macrophages were plotted against the absorbance values for each of the antigens. The values for CRIII expression did not differ significantly between resident macrophages, macrophages activated with recombinant interferon-gamma (rIFN-gamma) and macrophages activated with BCG/PPD. IFN-gamma-activated and BCG/PPD-activated macrophages expressed Ia antigen significantly more intensely than did resident peritoneal macrophages. In contrast the activated macrophages expressed F4/80 significantly less intensely than resident peritoneal macrophages.
Nibbering
P H
PH
Department of Infectious Diseases, University Hospital, Leiden, The Netherlands.
Van de Gevel
J S
JS
Van Furth
R
R
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D002448:Cell Adhesion; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D008262:Macrophage Activation; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D011951:Receptors, Complement / Q000032:analysis
D000954:Antigens, Surface; D000949:Histocompatibility Antigens Class II; D011951:Receptors, Complement