Typing of a panel of soluble HLA class I antigens by enzyme-linked immunosorbent assay.
Two ELISA assays were developed to test the reactivity of soluble sHLAs with anti-HLA class I mAbs of IgG or IgM isotype. A panel of 40 different alleles of sHLA antigens was produced using 57 lymphoblastoid B-cell lines, which had been generated from class-I-phenotyped PBLs. Using 14 mAbs, the expression of 13 different sHLA antigens (sHLA-A2, -A3, -A11, -A24, -A29, -B7, -B8, -B13, -B14, -B27, -B44, -B57, and -B58) by 43 different cell lines was confirmed. In addition, the expected absence of these alleles from the culture supernatant of 11 cell lines was confirmed. Cross-reactivities of mAbs observed by microlymphocytotoxicity assays were also detected by ELISA. The results of this extensive analysis confirmed previous results demonstrating that sHLA class I typing by ELISA correlated with HLA class I cell typing by microlymphocytotoxicity [1-3]. Furthermore, additional information about the fine specificities of two mAbs was obtained. An anti-B27/44 IgM mAb appeared to react only with sHLA-B44 but not with sHLA-B27; mAb CR11-351, previously reported to react with HLA-A2, 28, bound also to sHLA-A1, -3, -11, and -A24.
Pouletty
C
C
SangStat Medical Corporation, Menlo Park, CA 94025.
Mercier
I
I
Glanville
L
L
Tomavo
N
N
Igoudin
L
L
Pouletty
P
P
Buelow
R
R
D000483:Alleles; D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000276:immunology; D002472:Cell Transformation, Viral; D003601:Cytotoxicity Tests, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000032:analysis*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D016130:Immunophenotyping; D012995:Solubility
D000911:Antibodies, Monoclonal; D015395:Histocompatibility Antigens Class I; D007074:Immunoglobulin G; D007075:Immunoglobulin M