title: Summary report from the first international workshop on soluble HLA antigens. Paris, August 1992.
abstract: 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.
authors.affiliation: SangStat Medical Corporation, Menlo Park, California.
authors.forename: P
authors.initials: P
authors.lastname: Pouletty
authors.forename: S
authors.initials: S
authors.lastname: Ferrone
authors.forename: F
authors.initials: F
authors.lastname: Amesland
authors.forename: N
authors.initials: N
authors.lastname: Cohen
authors.forename: U
authors.initials: U
authors.lastname: Westhoff
authors.forename: D
authors.initials: D
authors.lastname: Charron
authors.forename: R M
authors.initials: RM
authors.lastname: Shimizu
authors.forename: H
authors.initials: H
authors.lastname: Grosse-Wilde
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D006680:HLA Antigens