Summary report from the first international workshop on soluble HLA antigens. Paris, August 1992.
The First International Workshop on Soluble HLA antigens focused on the comparison of immunoassay procedures for quantitation of soluble HLA (sHLA) class I antigens and the selection of a sHLA class I antigen international standard. Several sets of serum, plasma, and cell culture supernatant specimens were assayed blindly for levels of sHLA class I antigens by 15 participating laboratories using different immunoassay formats. The sandwich ELISA using (i) for antigen capture: an anti-HLA class I heavy chain monoclonal antibody (mAb) specific for a monomorphic epitope, and (ii) for antigen detection: an anti-beta 2 microglobulin antibody-enzyme conjugate, was the assay format of choice. There was a high inter-laboratory correlation among the majority of laboratories. All serum and plasma specimens from normal donors, and from a single transplant patient, had detectable levels of sHLA class I antigens. Paired serum and plasma specimens had similar levels of sHLA class I antigens, although plasma sHLA antigens seemed more stable than serum sHLA antigens. sHLA-A2 and sHLA-B7 antigens were detected in all specimens from HLA-A2 and HLA-B7 donors, respectively, using allele-specific ELISAs. No difference in reactivity was observed for quantitation of native sHLA class I antigens whether the capture mAb was TP25.99 (alpha 3 domain-specific) or W6/32 (alpha 2 + alpha 3-specific). However, a human-mouse chimeric sHLA class I antigen reacted weakly in assays which used TP25.99 mAb. The wide variation among laboratories in their reporting of micrograms/ml units pointed to the need for an inter-laboratory standardization based on a calibrated sHLA antigen preparation. T.sB7, an sHLA-B7 antigen derived from a cell line transfected within human beta 2 microglobulin and HLA-B7 genes, was accepted as the First sHLA class I Antigen International Standard at the workshop meeting.
Pouletty
P
P
SangStat Medical Corporation, Menlo Park, California.
Ferrone
S
S
Amesland
F
F
Cohen
N
N
Westhoff
U
U
Charron
D
D
Shimizu
R M
RM
Grosse-Wilde
H
H
D000483:Alleles; D000818:Animals; D000911:Antibodies, Monoclonal; D002678:Chimera; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005602:France; D006680:HLA Antigens / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007753:Laboratories; D051379:Mice; D010641:Phenotype; D012015:Reference Standards; D012995:Solubility; D011795:Surveys and Questionnaires
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006680:HLA Antigens