A flow cytometry-based method for detecting antibody responses to murine cytomegalovirus infection.
An assay based on target cells infected with green fluorescent protein labeled murine cytomegalovirus (GFP-MCMV) and dual color flow cytometry for detecting antibody to MCMV is described. After optimizing conditions for this technique, kinetics of anti-MCMV IgG antibody response was tested in susceptible (BALB/c) and resistant (C57BL/6) mouse strains following primary MCMV infection. Previously published antibody kinssetics were confirmed in susceptible mice, with peak IgG response seen approximately 8 weeks after primary infection, decreasing by 20 weeks after infection. In contrast, MCMV resistant C57BL/6 mice showed significantly lower IgG antibody responses than susceptible mice. Although several techniques have been previously described to detect murine antibody responses to MCMV, including nuclear anti-complement immunofluorescence, viral immunoblotting, complement fixation, indirect immunofluorescence, indirect hemagglutination, and enzyme-liked immunosorbent assay techniques, these techniques are all time consuming and laborious. The technique presented is a simple time efficient alternative to detect previous MCMV antibody responses in experimentally infected mice.
Bickerstaff
Alice A
AA
Departments of Surgery, The Ohio State University, Columbus, OH 43210, USA.
Zimmerman
Peter D
PD
Wing
Bret A
BA
Taylor
Frederick
F
Trgovcich
Joanne
J
Cook
Charles H
CH
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006566:Herpesviridae Infections / Q000276:immunology* / Q000821:virology; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018146:Muromegalovirus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D041681:NIH 3T3 Cells / Q000821:virology; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D049452:Green Fluorescent Proteins