Antibodies to nuclear proteins in cervical cancer. The Pap smear, an important clinical screen for uterine cervical cancer, has limited prognostic value for individual patients. The staging of cervical intraepithelial neoplasia (CIN) by cytological alone is often subjective. The recent correlation of human papilloma virus (HPV) types 16 and 18 with cervical carcinoma has added to the difficulty of making correct prognoses from Pap smears. We have fractionated Pap smear samples and purified nuclear matrix proteins from normal patients and from patients with carcinoma in situ. Nuclear matrix proteins are low abundance nonhistone proteins that are involved in nuclear structure and some aspects of the regulation of gene expression. Analysis of Pap smear nuclear matrix proteins by two-dimensional gel electrophoresis demonstrates that the pattern of expression of these proteins is diagnostic of each patient class. Several proteins are expressed specifically in normal or tumor- bearing patients. Monoclonal antibodies that react with these diagnostic proteins can be used with Pap smear samples to dramatically improve the prognostic accuracy of the test. We propose to characterize and develop monoclonal antibodies to several of these proteins. In this proposal we will: 1. Prepare and analyze nuclear matrix proteins from a clinically significant number of patients. We will seek to establish the diagnostic accuracy of nuclear matrix protein expression in normal and CIN III patients. We will extend this analysis to Pap smears from patients with mild-moderate dysplasias (CIN grades I and II) as well as normal patients with HPV 16 and 18 infections. 2. Eight proteins, initially selected for diagnostic potential from two-dimensional gel analysis, will be microsequenced. From the peptide sequence, synthetic peptides will be made in quantities sufficient for immunization and monoclonal antibody production. 3. Antibodies will be assayed immunohistochemical techniques using cervical cell suspensions. The clinical significance of an individual antibody will be determined by comparing it with the cytological diagnosis and with the progression of the disease. 4. The role of diagnostic nuclear matrix proteins in HPV infected patients of various CIN grades will be determined by simultaneous immunohistochemical assay and in situ hybridization of specific HPV types. The goal of these studies is to produce diagnostic antibodies that, when used in conjunction with conventional Pap smears, will reduce the subjectivity and improve the prognostic accuracy of the test.