AUTOREACTIVE EPITOPES WITHIN THE HUMAN ALPHA-ENOLASE AND THEIR RECOGNITION BY SERA FROM PATIENTS WITH ENDOMETRIOSIS

Patients with endometriosis significantly develop autoantibodies direc ted against endometrial proteins, which may be involved in the aetiolo gy of this gynaecological disease. Based on standard Western blot anal ysis, a 48 kDa protein was localized in the soluble protein extract of endometrial adenocarcinoma cells using sera from patients with clinic ally staged endometriosis and identified as the glycolytic enzyme alph a-enolase. The corresponding cDNA coding for the human alpha-enolase w as isolated from a human endometrial cDNA library and cloned into the vector pH6EX3, allowing the efficient expression of recombinant human alpha-enolase with an N-terminal histidine-hexapeptide as affinity Lig and in Escherichia coli. The purified recombinant human alpha-enolase was evaluated as a specific antigenic tool for the diagnostic measurem ent of antiendometrial antibodies in sera from patients with endometri osis. With selected endometriosis sera, two linear autoreactive epitop es were localized within the recombinant human alpha-enolase using epi tope mapping techniques, and they were characterized. (C) 1995 Academi c Press Limited