Sera obtained at enrollment in the study from patients suffering from moderate to sever dysplasia (cervical intraepithelial neoplasia grade II), carcinoma in situ (cervical intraepithelial neoplasia grade III) and invasive carcinoma, or developing any of these conditions in the course of the prospective study, and from control subjects, were examined for herpes simplex type-2 (HSV-2) antibody presence. The controls were matched with the patients by age, age at first intercourse, number of sexual partners, smoking habits and history of diathermoelectrocoagulation of the ectopic epithelium and transformation zone of cervix. Only those subjects were selected as controls who remained free of pathological colposcopical and cytological findings throughout the observation period, i.e. for at least 4 years after their serum sample was obtained. The microneutralization test (MNT) and type-2-specific solid-phase radioimmunoassay (SPRIA) were used as serological tests. No difference in the prevalence of HSV-2 antibody between the patients and controls was revealed by either test. Various combinations of the results from the two tests also failed to show any difference between patients and controls. Moreover, no significant differences were observed in the prevalence of HSV-2 antibody between patients suffering from the various pathological conditions and those diagnosed at enrollment and later in the course of the study. These results do not provide any support for the hypothesis of the involvement of HSV-2 in cervical neoplasia.

To determine the risk associated with previous herpes simplex type-2 (HSV-2) infection and possibly other virus infections, a prospective study of cervical neoplasia in more than 10,000 women was performed in the 1975-1983 period. The subjects were selected at random from an alphabetical listing of eligible women living in one district of Prague. At enrollment colposcopy and cervical cytology were performed, a blood sample was taken and data regarding education, socio-economic status, personal habits and sexual and reproduction-associated attributes were obtained from each woman. A total of 10,683 women were enrolled; a complete set of data was obtained in 10,389 women. Women with normal or non-significant findings were invited for further colposcopical and cytological investigations after 2 years and 4 years, the other women were followed at 3- to 6-monthly intervals. In women with highly significant findings, histological investigation was performed. The total of 150 cases of moderate to severe dysplasia (i.e. cervical intraepithelial neoplasia, grade II, CIN II), 83 cases of carcinoma in situ (CIN III) and 21 cases of invasive carcinoma (INCA) were detected. More than 60% of the patients were ill at enrollment, the other cases developed in subjects with originally slightly suspicious (27 CIN II, 17 CIN III, 3 INCA) or negative findings (30 CIN II, 12 CIN III, 3 INCA). Analysis of the data indicated significantly positive correlation of one or more of these clinical conditions with a number of sexual and reproduction-related attributes of which early age at first intercourse was most consistent. Among the other attributes, the smoking habit was associated with the highest risk of developing the disease. A negative correlation of cervical neoplasia with several attributes was demonstrated; of these diathermoelectrocoagulation of the ectopic epithelium and transformation zone of cervix was the most important single protective factor. On the basis of these findings, control subjects were selected for serological studies.

The entire amino acid sequence of human cytomegalovirus (HCMV) 150K matrix phosphoprotein (pp150), consisting of 1048 amino acid residues, was divided into 95 overlapping 20 amino acid peptides which were synthesized on polyethylene rods. The rods were subjected to ELISA with pooled anti-HCMV-positive and anti-HCMV-negative sera. Four peptides recognized by the anti-HCMV-positi~,e pool only were synthesized by the solid-phase method and their reactivity in a conventional ELISA, using a panel of 14 individual anti-HCMV-negative and 20 anti-HCMVpositive antisera, was evaluated; three peptides were found to be specifically reactive. Results obtained with one of these peptides (residues 595 to 614) in ELISA showed a good correlation with those obtained using a routinely performed complement fixation test.

Sera from inlfectious mononucleosis patients aged 2 to 35 years and from normal subjects aged I I to 35 years were examinedfor the presence of antibody to the EB virus capsid antigen ( V C A ) Extracts of Burkitt lymphoma and other lymphoblastoid cell lines contain a soluble (S)antigen demonstrated by complement-fixation (CF) test (Armstrong P I ul., 1966;Vonka et al., 1969;1970a; 1970b;Pope et al., 1969;Gerber and Diehl, 1970). This antigen was found in both EB-virus-positive and -negative cell lines and was immunologically distinct from the viral capsid antigen (VCA) as detected by immunofluorescence (IF) or CF test. The S antibody, although present in many human sera, was never found in sera which were free of antibody to VCA. This indicates that EB virus infection is the pre-requisite for the S antibody development. On the basis of these observations the suggestion was made that the S antigen is an EB virus coded non-structural antigen.When studying the age distribution of VCA and S antibodies in human sera we found that 50 to 70% of subjects over 1 year of age, whose sera were VCA-positive, also possessed the S antibody. The antibody pattern was markedly different in children aged 6 to 12 months: only one out of 15 VCA-positive subjects in one group (Vonka et al., 1970a) and none of 30 VCApositive in the other group (Vonka and BenyeshMelnick, 1970) possessed the S antibody. The absence of S antibody in VCA-positive subjects aged 6 to 12 months, i.e. subjects most probably infected with EB virus only recently,

Several vaccinia virus recombinants inducing the synthesis of the middle surface (M) protein of hepatitis B virus (HBV) were constructed. One of them, denoted v137, was examined in some detail. The virus replicated nearly to the same extent in various cell lines, viz. human embryo diploid fibroblast LEP and MRC-5 cells, rabbit embryo fibroblast REF cells, TK- rat RAT-2 cells, and green monkey CV-1 cells. However, the production of M protein was found considerably lower in the human LEP and MRC-5 than in the other cells examined. In addition, the kinetics of M formation were different in these two cell systems, LEP cells lagging significantly behind CV-1 cells. The low-level production of M protein in LEP cells was not increased by repeated v137 passages in LEP cells, nor by a passage in a laboratory worker accidentally infected with the v137 virus, nor by shortening the leader sequence preceding the translation initiation codon. The greater part of the M antigen was found to be cell associated, more so in the cells of human than monkey origin. From the major HBV S antigen (HBsAg) isolated from the plasma of chronically infected subjects, the antigen released by cell destruction differed by binding to polymerized human albumin. This property was utilized in ELISA to detect anti-preS2 antibody. Rabbits inoculated intradermally with the v137 virus developed antibodies reactive in this assay as well as with a synthetic peptide corresponding in the amino acids 14-34 of the NH2 terminus of the HBsAg preS2 region.