Identification, characterization, and quantitation of soluble HLA antigens in the circulation and peritoneal dialysate of renal patients.
Human lymphocyte antigen (HLA) class I and class II antigens and beta 2 microglobulin (B2M) were identified in peritoneal dialysate (PD) and serum from patients with end-stage renal disease (ESRD) using monoclonal antibodies in an enzyme-linked immunoassay. The HLA class I and class II antigens each exhibited approximate molecular weights of 50,000 to 60,000 daltons by chromatography on Sepharose CL 6B. Class I antigens in serum and PD fluid were associated with B2M. Free B2M (Mr 11,500) also was detected in both sera and PD fluids. Unlike class I antigens, class II antigens were not found to have attached B2M. Class I and class II antigens eluted from 2-diethylaminoethanol ion exchange gradient columns at 0.07 mol/L (molar) phosphate buffer pH 7.2 and migrated with alpha 2-beta 1 mobility in agarose electrophoresis. Class I antigens were purified from ESRD patients' PD fluid by solid-phase immunoaffinity chromatography. Enzyme-linked immunoassay demonstrated that this purified protein was composed of a class I heavy chain and B2M. Class I allospecificity was confirmed by neutralization on known HLA typing antisera in a microcytotoxicity assay. Soluble HLA class I antigen preparations specifically inhibited blast transformation of responder lymphocytes in mixed lymphocyte culture reactions. Inhibition was dose dependent and ranged from 0% to 95%. The presence of soluble HLA antigens in body fluids may play an important part in the immunologic tolerance to self. This study demonstrates a ready source of large quantities of soluble HLA for detailed analysis.
Gelder
F B
FB
Department of Surgery, Louisiana State University School of Medicine, Shreveport.
McDonald
J C
JC
Landreneau
M D
MD
McMillan
R M
RM
Aultman
D F
DF
D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D015314:Dialysis Solutions / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006680:HLA Antigens / Q000097:blood* / Q000737:chemistry; D006684:HLA-DR Antigens / Q000097:blood; D015395:Histocompatibility Antigens Class I / Q000097:blood* / Q000737:chemistry; D006801:Humans; D007108:Immune Tolerance; D015151:Immunoblotting; D007674:Kidney Diseases / Q000097:blood* / Q000276:immunology; D009500:Neutralization Tests; D010530:Peritoneal Dialysis; D012995:Solubility; D001613:beta 2-Microglobulin / Q000032:analysis / Q000378:metabolism
D015314:Dialysis Solutions; D006680:HLA Antigens; D006684:HLA-DR Antigens; D015395:Histocompatibility Antigens Class I; D001613:beta 2-Microglobulin