The use of flow cytometry to detect the biosynthesis of complement components.
Agarose beads, an activator of complement, were incubated with MRC-5 or He 9 fibroblast cell lines under serum-free conditions. The beads were tested for binding of anti-complement antibodies by flow cytometry with a FACS 440 using FITC-labelled anti-Ig detection antibodies. Controls consisted of co-cultured beads incubated with irrelevant antibody or albumin, beads maintained in cell cultures containing cycloheximide, and beads which were not exposed to cells. The histograms demonstrated positive staining with anti-C3c, -C5, -C7 and -C9, but not with anti-C6 and -C8. Flow cytometry with multiple histogram analysis confirmed that the differences between the positive curves and the controls were statistically significant. The results show that cell-derived complement components (C3, C5, C7 and C9) were deposited on the beads and could be detected by flow cytometry.
Hetland
G
G
Blood Bank, Ullevål Hospital, Oslo, Norway.
Garred
P
P
Mollnes
T E
TE
Størvold
G
G
D002460:Cell Line; D003165:Complement System Proteins / Q000096:biosynthesis*; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007118:Immunoassay / Q000379:methods; D066298:In Vitro Techniques; D012685:Sepharose
D003165:Complement System Proteins; D012685:Sepharose