Rosette formation between immobilised human lymphocytes and erythrocytes sensitised with monoclonal anti-D.
A rapid, reproducible and sensitive assay was developed to investigate the ability of human lymphocytes to form rosettes with erythrocytes sensitised with human monoclonal anti-D. Erythrocytes sensitised with a known number of anti-D molecules per cell were incubated with lymphocytes immobilised on plastic by poly(L-lysine), the resulting rosettes fixed, unbound erythrocytes removed by washing and the cell preparation stained. IgG1 and IgG3 anti-D-coated erythrocytes gave similar rosette formation at sensitisation levels in the range of 5000-15,000 molecules per cell, although at lower sensitisation levels IgG3 gave greater rosette formation than IgG1. A minimum of 500 IgG3 and 1000 IgG1 anti-D molecules per erythrocyte were required for rosetting.
Leader
K A
KA
United Kingdom Transplant Service, Southmead, Bristol.
Kumpel
B M
BM
Bradley
B A
BA
D000911:Antibodies, Monoclonal; D004912:Erythrocytes / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000276:immunology*; D012204:Rh-Hr Blood-Group System / Q000276:immunology*; D012397:Rosette Formation
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D012204:Rh-Hr Blood-Group System