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KMID : 0350519940470020887
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Journal of Catholic Medical College
1994 Volume.47 No. 2 p.887 ~ p.897
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Immunologic Diagnosis Using Human Papillomavirus Recombinant Proteins in the Patients of Cervical Cancer
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Abstract
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Host factors are important in determining the outcome of genital human papillomavirus (HPV) infection as demonstrated by an increase in cervical cancers in immunosuppressed patients. HPV proteins may pepresent immunological target such as serum
antibodies against HPV infection. However, evaluation of the antibody response in patients exposed to high-risk HPV types has been hampered by the lack of a readily available source of viral proteins for use as target antigens in serologic
assays.
The
goals of the study presented here aro to determine whether serum antibodies recognized HPV fusion proteins, to identify which antigens are targets of the humoral immune response, and to assess the prevalence of HPV antibodies in defined
populations. In
this study, the assay was tested on selected group of sera from the patients with carcinomas (n=81), squamous intraepithelial lesion (SIL) (n=25) of uterine cervix and normal controls (n=40) using the purified TrpE fusion open reading frame (ORF)
proteins of HPV-16 by Western blot immunoassay.
@ES The results of our experiments were
@EN 1) Restriction enzyme fragments from ORFs of HPV-16 were expressed in E. Coli as tryptophan E synthetass-HPV fusion proteins through the expression vector pATH. Each fusion protein was induced by 3-indoleacrylic acid and was purified from the
bacterial lysates for polyacrylamide gel electrophoresis and Western blot immunoassay.
2) Among women with cervical cancer, 15/18 (19%) and 54/81(19%) and 54/81 (67%) were positive for antibodies by Western blot immuncassay to any gene products of HPV-6b and HPV-16 respectively. And in 25 patients with HPV-related SIL, 10 (40%)
and
11
(44%) were positive for antibodies against HPV-16 related proteins respectively. The positive prevalence rate for HPV-6b antibodies in the sera of SIL patients were significantly higher than that of in the sera of cervical cancer patients
(P<0.05).
The
prevalence rates of antibodies to ORF proteins of HPV-16 in the sera of cervical cancer patients were 12% (10/81 ; E1), 10% (8/81 ; E2), 21% (17/18 ; E4), 33% (27/81 ; E6), 36% (29/81 ; E7), 37% (30/81 ; L1), 11% (9/81 ; L2) respectively, while
8%
(2/25) were for E1, 8%( 2/25) for E2, 12% (3/25) for E4, 40% (10/25) for E6, 32% (8/25) for E7, 28% (7/25) for L1, and 8% (2/25) for L2 protein among the patients with HPV-related cervical SIL. The positive prevalence rates for HPV-16 antibodies
in
the
sera of SIL and cervical cancer patients were significantly higher than those of in the sera of normal control s(p<0.01).
3) There was decreasing pattern of positivity against E6 protein of HPV-16 with the severity of cervical cancer, but there were no significant differences in positivities of other antibodies in the patients of cervical cancers according to the
different stages of disease.
These results suggest that the recombinant proteins from HPV-6b and HPV-16 may be useful in the immunologic diagnosis of HPV-related cervical SILs and cancers respectively and antibodies to HPV-16 E7 and L1 proteins appear to be virus-specific
and
disease-specific markers for HPV-associated cervical neoplastic lesions.
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KEYWORD
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