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자궁경부암에서 HPV-16 단백질을 이용한 혈청내 진단물질의 임상적 효용성
김승조,남궁성은,박종섭,김현수,김찬주,박순희,안희경,정근택,박동춘,유왕돈 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.3
Background : Most markers in current clinical use are not cancer-specific, but the ideal tumor marker should be specific for cancer, highly sensitive, clinically practicable, inexpensive, and acceptable to individual. it is still debating that SCCA and CEA are clinically useful for specific tumor markers in management of cervical cancers. Recently, a variety of HPV- related proteins have been synthesized and their utility as diagnostic and prognostic tumor markers in cervical cancers is needed to be assessed by comparing the previous tumor markers ; CEA and SCCA. The early proteins(E6 & E7) of HPV-16 which are responsible for malignant transformation could be constructed by in vitro transcription and translation and the late structural proteins(L1/L2) of HPV-16 have been shown to self-assemble into virus-like particles(VLPs) from baculovirus system when expressed in insect cells. The ability to generate preparative amounts of HPV-16 E6, E7 proteins and L1/L2 VLPs may have implications for the development of a serologic assay to detect anti-HPV-16 virion immune responses to conformational epitopes of HPV-associated cervical neoplasia. Methods : We attempted to investigate serologic response in the sera obtained from Korean women with cervical neoplasia by RIPA(radioimmunoprecipitation assay) using in vitro translated HPV-16 E6, E7 proteins and ELISA using HPV-16 L1/L2 VLPs, PCR method using E6 type-specific primers and probes for HPV-16/18 was used to determine the presence and type of HPV infection in the study population(normal controls ; 15 cases, preinvasive lesions ; 28 patients, and invasive cervical cancers; 124 cases). Result : 1) The sera of 0% of preinvasive lesions and 34%(42/124) of cervical cancers were positive for SCCA and the sera of 0% of preinvasive lesions and 18% (22/124) of cervical cancers were positive for CEA. The positivity of SCCA was increased with advancing clinical stages, but the antibody levels were not correlated with clinical outcome of disease. 2) The sera of 7%(2/28) of preinvasive lesions and 51%(63/124) of cervical cancers were positive for in vitro translated HPV-16 E6 protein(P$lt;0.05) and the sera of 11%(3/28) of preinvasive lesions and 33%(41/124) of cervical cancers were positive for in vitro translated HPV-16 E7 protein(P$lt;0.05). The levels of HPV-16 E6 proteins showed high titers in some invasive cancers, but they didn`t show any fluctuations along with clinical outcomes. In contrast, the antibody levels to HPV-16 E7 protein were correlated to clinical stage and tumor burden in a significant number of cervical cancers. 3) The sera of 7% (1/15) of normal controls, 39%(11/28) of preinvasive lesions and 56% (70/124) of cervical cancers were ELISA positive for HPV-16 L1/L2 VLPs(P$lt;0.05). The positive reactivities for HPV-16 L1/L2 VLPs in cervical cancers by clinical stage, histopathology or HPV DNA types were not significantly different. Conclusion : These results suggest that the considerable number of patients with cervical neoplasia generated positive antibody response to in vitro translated HPV -16 E6, E7 proteins and L1/L2 VLPs. These HPV-16-associated proteins might be disease-specific markers which could be useful in adjunctive diagnostic assay and seroepidemiologic study of HPV-related cervical neoplasia. Especially, the monitoring of antibody to HPV-16 E7 protein seems to be valuable in proper management of cervical cancers for specific tumor marker.
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자궁경부암 환자에서 ELISA를 이용한 HPV-16 VLP에 대한 혈청반응
김승조,남궁성은,박종섭,김현수,김찬주,박순희,안희경,황은성,정근택 대한산부인과학회 1996 Obstetrics & Gynecology Science Vol.39 No.1
1993년 1월부터 1994년 12월까지 가톨릭대학교 의과대학 강남성모병원 산부인과 종양학부에서 진단 및 치료를 받았던 환자들에서 치료전 채취한 혈청과 자궁경부 세포를 이용한 실험결과 다음과 같은 결론을 얻을 수 있었다. 1. 자궁경부암 전구병변 환자 20명은 HPV-16에 양성이었고, 나머지 4명은 다른 아형의 DNA HPV가 있거나, 발견되지 않았다. 96명의 자궁경부암 환자에서 69명은 HPV-16 DNA, 9명은 HPV-18 DNA에 양성이었고, 그외 3명은 HPV-16과 HPV-18이 동시에 발견되었다. 또한 7명에서는 다른 아형의 HPV DNA가 있었고, 나머지 8명은 HPV DNA를 발견할 수 없었다. 2. HPV-16 L1, L2 VLP에 대한 ELISA 결과는 정상 대조군의 9%에서 양성이었고, 자궁경부 전구병변의 환자에서는 50%(12/24), 자궁경부암 환자의 62.5%에서 각각 양성으로 나타나 자궁경부의 종양성 변화가 있는 환자에서 유의한 진단적 가치가 있었다. 3. HPV-16 DNA에 양성인 자궁경부암 환자의 68%(47/69)에서 HPV-16 VLP ELISA에 양성이었고, HPV-18 DNA에 양성인 환자의 44%(4/9)에서 HPV-16 VLP ELISA에 양성이었으나 HPV-16/18이외의 다른 HPV 아형에 양성인 환자에서는 14%(1/7)에서 ELISA 양성으로 나타나서 HPV-16또는 HPV-18 DNA 양성인 환자들과 다른 종류 HPV DNA에 양성인 환자들 사이에서 항체 반응 여부는 통계학적인 차이가 있었다. 4. 자궁경부암 환자의 임상기와 조직학적 특성에 따른 HPV-16 VLP에 대한 항체 형성을 관찰해 본 결과 각 임상기와 조직형에 따른 차이를 관찰할 수 없었다. 이상의 결과로 자궁경부암과 전구 병변 환자들의 대다수에서 ELISA 방법으로 HPV-16 VLP에 대한 항체 형성을 관찰할 수 있었다. 또한 HPV-16 VLP ELISA 검사는 HPV-16 DNA 검사의 보조진단법으로 유용하며, HPV와 연관된 자궁경부 종양성 질환의 혈청학적 역학 연구에도 도움을 줄 수 있을 것으로 사료된다. Background: The late structural proteins of HPV-16 have recently been shown to self-assemble into virus-lide particles(VLPs) from baculovirus system when expressed in insect cells. The ability to generate preparative amounts of HPV-16 L1-L2 VLP may have implications for the development of a serologic assay to detect anti-HPV-16 virion immune responses to conformational epitopes and for immunoprophylasis against HPV-16 infection. Methods: We attempted to investigate serologic responses in the sera obtained from Korean women with cervical neoplasia by ELISA using HPV-16 VLPs. PCR method using L1 consensus primers and type-specific oligomer probes was used to determine the presence and type of HPV infection in the study population(normal control; 23 cases, preinvasive lesions; 24 cases, and invasive cervical cancers; 96 cases). Result: 1) The sera of 93%(21/23) of women, who didn`t have HPV-ralated cervical lesions, were negative for HPV-16 VLPs in ELISA assay. The sera of 50%(12/24) of patients with preinvasive cervical lesion and 63%(60/96) of patients with invasive cervical cancer were ELISA positive for HPV-16 VLPs(p$lt;0.01). 2) The positive reactivities for ELISA of HPV-16 VLPs in cervical cancer patients according to clinical characteristics were nor signific antly different. 3) Among women with cervical neoplasia, HPV 16 VLP was positive in 50%(10/20) of HPV-16 positive cases with preinvasive cervical lesion and 72%(50/72) of HPV-16 positive cases with invasive cervical cancer. Conclusion: These results suggest that the considerable number of patients with cervical neoplasia generated an IgG antibody response to the HPV 16 VLPs that can be detected by ELISA. These HPV-16 VLP ELISA may be useful in adjunctive diagnostic asay to HPV-16 DNA test and seroepidemiologic study of HPV-related cervical neopl-asia.
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