Binding by Immunoglobulin to the HPV-16-derived Proteins LI and E4 in Cervical Secretions of Women with HPV-related Cervical Disease1

Although DNA of the human papilloma viruses (HPV) can be identified in epithelium of a large proportion of patients with genital squamous lesions, relatively little is known about the extent of the local host immune response to this virus. We analyzed cervical secretions from patients undergoing evaluation because of abnormal Papanicolaou smears (cervi cal biopsy showed nonspecific atypia, flat condyloma, or intraepithelial neoplasia), as well as controls, for immunoglobulin binding to proteins produced in vitro to HPV-16 LI, E4, and E7 open reading frames. Segments of the HPV-16 genome, including portions of the LI (1111cleotides 6153-6794), E4 (nucleotides 3399-3648), and E7 (nucleotides 686-880) open reading frames, were cloned into pATH vectors and expressed as tryptophan synthetase E fusion proteins in Escherichia coli and used as a source of study antigens. Fusion proteins containing the HPV LI, E4, and E7 polypeptides were found to be distinct by molecular weight (59,000; 45,000; and 42,000) as well as by immunological deter minants recognized by heterologous immune sera. Of 8 cervical intraep ithelial neoplasia lesions tested by RNA-RNA in situ hybridization, 7 were found to be positive for HPV-16-related nucleic acids, in contrast to none (0 of 4) in the condyloma group (three positive for HPV DNA other than type 16). Immunoglobulin in cervical secretions showed reac tivity to HPV type 16 E4 or LI or both, with highest binding in patients with cervical intraepithelial neoplasia (P < 0.01 for HPV-16 LI and E4 compared with controls). Binding was not tryptophan synthetase E de pendent and was, in general, coincident for the HPV-16 E4 and LI proteins. We conclude that study of cervical secretions, using a quanti tative assay for immunoglobulin binding to HPV-16 proteins produced in vitro, may be useful to document the quality and quantity of the immune response of the host to this important human pathogen.