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Korean Journal of Obstetrics & Gynecology 1998;41(7):1865-1871.
Published online January 1, 2001.
Comparison of In Vitro Translated HPV-16 E7 Protein with GST-fusion HPV-16 E7 Protein in Clinical Utility as the Serologic Marker in the Patients with Cervical Cancer.
J S Park, T Y Kim, E J Kim, C J Kim, D C Park, S H Park, S J Kim, S E Namkoong, S J Um
Abstract
OBJECTIVE
Recently, a variety of HPV-related proteins have been synthesized and their utility as diagnostic and prognostic markers in cervical cancers is needed. The ability to generate preparative amounts of HPV-16 E7 proteins may have implications for the development of a serologic assay to detect anti-HPV-16 virion immune responses. The purpose of the study is to improve the way of proper management of the cervical cancer by investigating the clinical utility of in vitro translated HPV-16 E7 protein and GST-fusion HPV-16 E7 protein as the serologic marker through comparing the antibody reactions in the sera of the patients. METHODS: The serologic responses were investigated in Korean women with cervical neoplasia by radioimmunoprecipitation assay (RIPA) using in vitro translated HPV-16 E7 protein and ELISA using GST-fusion HPV-16 E7 protein. PCR using E6 type-specific primers for HPV-16/18 were used to determine the presence and type of HPV infection (normal controls; 15 cases, preinvasive lesions; 15 cases, and invasive cervical cancers; HPV-16 DNA positive 78 cases and HPV-16 DNA negative 22 cases). RESULTS: The sera of 39% (39/100) of patients with cervical cancer were RIPA positive for in vitro translated HPV-16 E7 protein [HPV-16 DNA positive cases; 41% (32/78), HPV-16 DNA negative cases; 32% (7/22)]. The sera of 29% (29/100) of cervical cancers were ELISA positive for HPV-16 GST-fusion E7 protein [HPV-16 DNA positive cases; 33% (26/78), HPV-16 DNA negative cases; 14% (3/22)]. By the two different methods, 12 cases were positively matched and 32 cases were negatively matched. The positivities for in vitrotranslated HPV-16 E7 protein in the patients with cervical cancer were 14% (7/49) in stage I, 59% (26/44) in stage II, and 86% (6/7) in stage III/IV, by significantly correlating with the advancing stage of the disease (P<0.01). CONCLUSIONS: These data suggests that the considerable number of patients with cervical neoplasia generated positive antibody response to in vitro translated protein and GST-fusion protein of HPV-16 E7. In contrast to RIPA method using in vitro translated protein, ELISA method using GST-fusion protein has several advantages for clinical application; use of non-isotope, easy preparation to produce a sufficient of protein, available large scale study. But the positive rates to in vitro translated HPV-16 E7 protein in the patients of cervical cancer were higher in general than those to GST-fusion HPV-16 proteins. Furthermore, the antibody positivities to in vitro translated HPV-16 E7 protein were increased by advancing clinical stage of disease. These HPV-16 E7 proteins might be the disease-specific markers which could be useful in adjunctive diagnostic assay and seroepidemiologic study of HPV-related cervical neoplasia.
Key Words: HPV-16 E7 in vitro translated protein, HPV-16 E7 GST-fusion protein, Cervical cancer, RIPA, ELISA
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