Analysis of HLA specificity of human monoclonal antibodies by cytofluorimetry and cell ELISA.
In this study we report on the characterization of cytotoxic human monoclonal antibodies (HmAb) detecting polymorphic HLA class II specificities using cytofluorimetric analysis in combination with micro cell ELISA. In both techniques, five anti-HLA HmAb were tested against HLA-transfected murine L cells as target cells and the bound antibody was detected, either by cytofluorimetry or by cell ELISA reader, after addition of fluoresceinated or peroxidase-conjugated anti-human IgG+IgM antibodies, respectively. The results demonstrate that HmAb directed against HLA-DR, -DQ and -DP molecules can be efficiently discriminated by cytofluorimetry and cell ELISA, which appear to be highly sensitive and perfectly comparable to the standard cytotoxicity assay.
Pistillo
M P
MP
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Tazzari
P L
PL
Mazzoleni
O
O
Urlacher
A
A
Falco
M
M
Vitale
M
M
Karr
R W
RW
Ferrara
G B
GB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D003601:Cytotoxicity Tests, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006681:HLA-D Antigens / Q000276:immunology*; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D006681:HLA-D Antigens; D011994:Recombinant Proteins