A flow cytometric rosetting assay for the analysis of Fc receptors and C3 receptors on HSV-infected cells.
A sensitive and reproducible flow cytometric assay was developed for the analysis of Fc gamma and C3b(i) receptors on HSV-infected cells. The method is based on a rosette technique using fluorochrome-labeled erythrocytes sensitized with IgG or C3b(i). A comparison of flow cytometric and microscopic quantitation demonstrated that the binding of EIgG, EC3b(i) to HSV-infected cells were correlated. Flow cytometric analysis provides the opportunity to study simultaneously the distribution of E per HSV-infected cell and the total binding of E to the whole population of HSV-infected cells. Receptor activity and HSV glycoprotein cell surface expression were shown to be correlated in a linear fashion. The assay could be applied to other Fc gamma R- and C3b(i)R-bearing cells.
Van Vliet
K E
KE
Eijkman-Winkler Laboratory of Medical Microbiology, Utrecht University, Netherlands.
De Graaf-Miltenburg
L A
LA
Verhoef
J
J
Van Strijp
J A
JA
D000818:Animals; D005434:Flow Cytometry; D006023:Glycoproteins / Q000032:analysis; D006561:Herpes Simplex / Q000276:immunology*; D006801:Humans; D051379:Mice; D008853:Microscopy; D017463:Receptors, Complement 3b / Q000032:analysis*; D011961:Receptors, Fc / Q000032:analysis*; D012397:Rosette Formation; D012756:Sheep; D014764:Viral Proteins / Q000032:analysis
D006023:Glycoproteins; D017463:Receptors, Complement 3b; D011961:Receptors, Fc; D014764:Viral Proteins