Antibody detection against HPV16 E7 & GP96 fragments as biomarkers in cervical cancer patients.
OBJECTIVE
Cervical cancer is the second most frequent cancer among females worldwide, especially human papilloma viruses (HPV) types 16 and 18. In viral systems the identification of serological markers would facilitate the diagnosis of HPV infections and virus-related disease. The aim of the present investigation was to determine and search for serologic markers in cervical cancer patients associated with HPV.


METHODS
A total of 58 Iranian women with invasive cervical carcinoma including adenocarcinoma and squamous cell carcinoma (SCC) were included. Serum antibody response to HPV infections in patients was detected by Western blot and ELISA techniques based on recombinant HPV16E7 and the N-terminal and C-terminal fragments of gp96 (NT-gp96 and CT-gp96) proteins. These recombinant proteins were expressed in Escherichia coli as a His-tag protein and purified using affinity chromatography.


RESULTS
The ELISA results indicated that patients with high antibody response to HPV16E7 had significant seroreactivity to CT-gp96 fragment. In Western blot analysis, a strong association between anti-E7, anti-NT-gp96 and anti-CT-gp96 reactivity and cervical cancer was obtained using purified recombinant proteins. In adenocarcinoma cases, no significant difference was observed in seroreactivities between normal and patients.


CONCLUSIONS
The evaluation of cervical cancer patients' seroreactivities against three recombinant proteins (rE7, rNT-gp96 and rCT-gp96) showed significantly higher levels of these markers in SCC only, but not in adenocarcinoma and control groups. Also, the usage of both techniques (ELISA and Western blotting) can provide more reliable tools for diagnosis of cervical cancer.
Bolhassani
Azam
A
Molecular Immunology & Vaccine Research Laboratory, Pasteur Institute of Iran, Tehran, Iran.
Zahedifard
Farnaz
F
Taslimi
Yasaman
Y
Taghikhani
Mohammad
M
Nahavandian
Bijan
B
Rafati
Sima
S
D000328:Adult; D000368:Aged; D000912:Antibodies, Neoplasm / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000276:immunology; D017931:DNA Primers / Q000235:genetics; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics / Q000276:immunology; D006801:Humans; D007492:Iran; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D008875:Middle Aged; D050725:Papillomavirus E7 Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D002583:Uterine Cervical Neoplasms / Q000276:immunology / Q000821:virology*; D055815:Young Adult
D000912:Antibodies, Neoplasm; D000914:Antibodies, Viral; D014408:Biomarkers, Tumor; D017931:DNA Primers; D008562:Membrane Glycoproteins; D050725:Papillomavirus E7 Proteins; D011994:Recombinant Proteins; C053387:endoplasmin; C059731:oncogene protein E7, Human papillomavirus type 16