Differential Antibody Response against Conformational and Linear Epitopes of the L1 Proteins from Human Papillomavirus Types 16/18 Is Observed in Vaccinated Women or with Uterine Cervical Lesions (original) (raw)
Related papers
Infectious agents and cancer, 2006
Human papillomaviruses (HPVs) are the primary etiological agents of cervical cancer and are also involved in the development of other tumours (skin, head and neck). Serological survey of the HPV infections is important to better elucidate their natural history and to disclose antigen determinants useful for vaccine development. At present, the analysis of the HPV-specific antibodies has not diagnostic value for the viral infections, and new approaches are needed to correlate the antibody response to the disease outcome. The aim of this study is to develop a novel ELISA, based on five denatured recombinant HPV16 proteins, to be used for detection HPV-specific antibodies. The HPV16 L1, L2, E4, E6 and E7 genes were cloned in a prokaryotic expression vector and expressed as histidine-tagged proteins. These proteins, in a denatured form, were used in ELISA as coating antigens. Human sera were collected from women with abnormal PAP smear enrolled during an ongoing multicenter HPV-Pathogen...
Infectious agents and …, 2006
BackgroundHuman papillomaviruses (HPVs) are the primary etiological agents of cervical cancer and are also involved in the development of other tumours (skin, head and neck). Serological survey of the HPV infections is important to better elucidate their natural history and to disclose antigen determinants useful for vaccine development. At present, the analysis of the HPV-specific antibodies has not diagnostic value for the viral infections, and new approaches are needed to correlate the antibody response to the disease outcome. The aim of this study is to develop a novel ELISA, based on five denatured recombinant HPV16 proteins, to be used for detection HPV-specific antibodies.MethodsThe HPV16 L1, L2, E4, E6 and E7 genes were cloned in a prokaryotic expression vector and expressed as histidine-tagged proteins. These proteins, in a denatured form, were used in ELISA as coating antigens. Human sera were collected from women with abnormal PAP smear enrolled during an ongoing multicenter HPV-PathogenISS study in Italy, assessing the HPV-related pathogenetic mechanisms of progression of cervical cancer precursor lesions. Negative human sera were collected from patients affected by other infectious agents. All the HPV-positive sera were also subjected to an avidity test to assess the binding strength in the antigen-antibody complexes.ResultsMost of the sera showed a positive reactivity to the denatured HPV16 proteins: 82% of the sera from HPV16 infected women and 89% of the sera from women infected by other HPV genotypes recognised at least one of the HPV16 proteins. The percentages of samples showing reactivity to L1, L2 and E7 were similar, but only a few serum samples reacted to E6 and E4. Most sera bound the antigens with medium and high avidity index, suggesting specific antigen-antibody reactions.ConclusionThis novel ELISA, based on multiple denatured HPV16 antigens, is able to detect antibodies in women infected by HPV16 and it is not genotype-specific, as it detects antibodies also in women infected by other genital HPVs. The assay is easy to perform and has low cost, making it suitable for monitoring the natural history of HPV infections as well as for detecting pre-existing HPV antibodies in women who receive VLP-based HPV vaccination.
International Journal of Cancer, 1992
Antibodies to the major (LI) coat protein of human papillomavirus type 16 (HPV-16) in sera from patients with cervical intra-epithelial neoplasia (CIN) have been investigated by means of recombinant proteins and synthetic peptides. When LI-HPV-I6 fusion proteins were used in immunoblot assays, no antibody reactivity was found in sera from 52 patients with CIN or from 2 I unrelated children. Amino-acid sequence analyses indicated that L I -HPV-I 6 amino acids 473 to 492 may contain an HPV-16 type-restricted epitope since the greatest diversity occurs in this region. In the ELISA, seropositivity to peptides 473 to 492 was more common among CIN patients whose biopsies contained HPV-I 6 DNA (9 I Yo, 2 I of 23) than among their children (24%, 5 of 2 I ; p < 0.00 I) or other CIN patients with HPV-I 6 DNA-negative biopsies (66Y0, I9 of 29; p < 0.05), but was unrelated to the severity of the CIN lesion. Antibodies to LI-HPV-16 peptide 473 to 492 among seropositive CIN patients cross-reacted with the analagous LI -HPV-33, but not with the LI-HPV-6b peptide, and were predominantly IgM. In contrast, antibodies which recognized a less variable region of L I -HPV-I 6 (amino acids 279 to 293) showed no association with HPV-I 6 DNA status. Seropositivity to the L I -HPV-6b (amino acids 473-492) was less frequent (33%) among CIN patients and unassociated with HPV-I 6 DNA status (p > 0. I); however
Clinical and Vaccine Immunology, 2008
We have very limited information on serum neutralizing antibody in women naturally infected with the human papillomaviruses (HPVs) that are causally associated with cervical cancer. In this study, serum samples collected from 217 Japanese women with low-grade cervical intraepithelial neoplasia were examined for their neutralizing activities against HPV16, -18, -31, -52, and -58 pseudovirions. Eighty-four patients (39%), 35 patients (16%), 17 patients (8%), and 1 patient were positive for neutralizing antibodies against one, two, three, and four of these types, respectively. Presence of neutralizing antibody did not always correlate with detection of HPV DNA in cervical swabs collected at the time of blood collection. The neutralizing titers of the majority of sera, ranging between 40 and 640, were found to be conserved in the second sera, collected 24 months later, independently of emergence of HPV DNA in the second cervical swabs. The data strongly suggest that HPV infection induce...
Journal of Medical Virology, 1995
Antibodies against eight synthetic peptides spanning different epitopes located on L1, L2, and E4 proteins of human papillomavirus (HPV) types 16,6, and 11 were examined in sera from 73 women infected by HPV and from 139 healthy controls. Only three of these peptides were reactive. Two located on proteins L2 and E4 of HPV 16 seem type specific since antibodies to these peptides were detected, respectively, in 21% and 15% of the HPV 16 infected patients and in 2.5% and none of women infected by other HPVs. The third peptide located on the L1 protein of HPV 6 bears a common epitope since antibodies to this peptide were detected not only in 85% of women infected by HPV 6 or 11, but also in 82% of women infected by other HPVs, and in 74% and 71% of the control groups (lO-l2-year-old children and adults, respectively). In conclusion, none of the peptides investigated seems useful to develop ELlSAs for serological diagnosis of HPV infection. o 1995 WiIev-Liss, Inc.
2014
Background: Human papillomavirus (HPV) infection involved in the development of more than 90% of cases of cervical cancer. Therefore, the aim of this study was to assess role of IHC for detection of HPV subtypes 16 and 18, as compared to molecular identification. Methodology: A total of 84 cervical cancer tissues that were previously diagnosed as having cervical cancer, were investigated for the presence of HPV subtypes 16 and 18. Both immunohistochemistry and molecular (Polymerase Chain Reaction (PCR)). Results: In application of molecular (PCR) testing for HPV subtypes 16 and 18; 62/84 (73.8%) were found positive for HPV subtype 16 and the remaining 22/84 (26.2%) were negative. On the other hand, on testing the samples for HPV subtype 18; 23/84 (27.4%) were found positive and the remaining 61/84 (72.6%) were found negative. Conclusion: Immunohistochemistry using clone K1H8 (anti-HPV) is considerably lower-priced, and has reasonable specificity to be applied in Yemen for screening for HR_HPV.
Medicine, 2016
Cervical cancer (CC) is the second most frequent neoplasia among women worldwide. Cancer prevention programs around the world have used the Papanicolaou (Pap) smear as the primary diagnostic test to reduce the burden of CC. Nevertheless, such programs have not been effective in developing countries, thus leading to research on alternative tests for CC screening. During the virus life cycle and in the process toward malignancy, different human papillomavirus (HPV) proteins are expressed, and they induce a host humoral immune response that can be used as a potential marker for different stages of the disease. We present a new Slot blot assay to detect serum antibodies against HPV16 E4, E7, and VLPs-L1 antigens. The system was validated with sera from a female population (n ¼ 485) aged 18 to 64 years referred to the dysplasia clinic at the General Hospital in Cuautla, Morelos, Mexico. To evaluate the clinical performance of the serological markers, the sensitivity, specificity, positive, and negative predictive values and receiver-operating characteristic curves (for antibodies alone or in combination) were calculated in groups of lesions of increasing severity. The results showed high prevalence of anti-E4 (73%) and anti-E7 (80%) antibodies in the CC group. Seropositivity to 1, 2, or 3 antigens showed associations of increasing magnitude with CC (odds ratio [OR] ¼ 12.6, 19.9, and 58.5, respectively). The highest association with CC was observed when the analysis was restricted to only anti-E4þE7 antibodies (OR ¼ 187.7). The best clinical performance to discriminate CC from cervical intraepithelial neoplasia 2 to 3 was the one for the combination of anti-E4 and/or anti-E7 antibodies, which displayed high sensitivity (93.3%) and moderate specificity (64.1%), followed by anti-E4 and anti-E7 antibodies (73.3% and 80%; 89.6% and 66%, respectively). In addition, the sensitivity of anti-E4 and/or anti-E7 antibodies is high at any time of sexual activity (TSA), which suggests they can be biomarkers for the early detection of CC. The sensitivity of anti-E4 antibodies was low (<10%) when the TSA was <10 years, and it increased up to 100% in relation to the TSA, suggesting that anti-E4 antibodies can be useful as HPV exposure markers at early stages of the disease. (Medicine 95(6):e2769) Abbreviations: AU = arbitrary units, AUC = area under the receiveroperating characteristics curve, CC = cervical cancer, CI = confidence interval, CIN = cervical intraepithelial neoplasia, ELISA = Enzyme Link Immunosorbent Assay, GFP = green fluorescent protein, HPV = human papillomavirus, HR = high risk, NPV = negative predictive value, OR = odds ratio, Pap = Papanicolaou, PBS = phosphae buffer solution, PPV = positive predictive value, TSA = time of sexual activity, VLPs = virus-like particles.
Journal of General Virology, 2008
We investigated neutralizing antibodies to human papillomavirus type 16 (HPV-16) in serum and cervical washes from 84 women with normal cytology or cervical disease. Serum neutralizing antibodies were detected in 78 % of women infected at the cervix with HPV-16, compared with 35 % (P50.002) of women infected with HPV-16-related types (a9 HPV types), 14 % (P,0.0001) of women infected with HPV-16 non-related types and none of HPV-uninfected women. A significant correlation between HPV-16 infection and serum HPV-16-neutralizing antibodies was observed (r s 50.97; P50.032). Cervical neutralizing antibodies were detected in 38 % of women with HPV-16 infection and in 17 % of women infected with the HPV-16-related type HPV-31. Cervical neutralizing antibodies correlated with HPV-16 infection (r s 50.95; P50.08), but not with cervical disease. Serum and cervical HPV-16 antibody responses were not affected significantly by human immunodeficiency virus type 1 infection. In conclusion, serum and cervical HPV-16-neutralizing antibodies were found to correlate with HPV-16 infection, but not with cervical disease.
Cancer Research, 2006
Oncogenic human papillomavirus (HPV) infection is a necessary cause of cervical cancer. Therefore, vaccination to prevent or eliminate HPV infection could reduce the incidence of cervical cancer. A fusion protein comprising HPV16 L2, E6, and E7 is a candidate combination preventive and therapeutic HPV vaccine. The L1-and L2-specific and neutralizing serum antibody titers and peripheral blood mononucleocyte antigen-specific proliferative responses generated by vaccination thrice at monthly intervals with HPV16 L2E7E6 were compared in two studies: a phase I randomized double-blind placebo controlled dose escalation trial in 40 healthy volunteers and a phase II trial of HPV16 L2E7E6 at the maximum dose in 29 women with highgrade anogenital intraepithelial neoplasia (AGIN). Vaccination of healthy volunteers induced L2-specific serum antibodies that were detected 1 month after the final vaccination (P binomial < 0.001). There was a significant trend to seroconversion for HPV16 and HPV18 neutralizing antibodies with increasing vaccine dose (P = 0.006 and P = 0.03, respectively). Seroconversion for HPV18 neutralizing antibodies showed a significant positive trend with increasing dose (P = 0.03) and was associated with seroconversion for HPV16 neutralizing antibodies (P exact = 0.04). The antigen-specific proliferative response of vaccinated healthy volunteers also showed a significant trend with increasing vaccine dose (P = 0.04). However, AGIN patients responded less effectively to vaccination than healthy patients for induction of HPV16 L2-specific antibody (P < 0.001) and proliferative responses (P < 0.001). Vaccination of healthy volunteers thrice with 533-Mg HPV16 L2E7E6 at monthly intervals induced L2specific serum antibodies that neutralized across papillomavirus species. Responses in AGIN patients were infrequent. (Cancer Res 2006; 66(23): 11120-4) Materials and Methods Human sera. Studies were done with the permission of the Johns Hopkins University Internal Review Board. The study sera were obtained in two HPV16 L2E7E6 vaccination trials from 40 healthy volunteers (30 men, ages 19-55 years, and 10 postmenopausal women, ages 43-55 years, with a negative Pap smear at entry) at weeks 0 and 12 in a phase I trial (12) and from 27 women, ages 22 to 61 years, all with noncervical high-grade AGIN (2 of 27 with VAIN3 and 25 of 27 with VIN3, all but two being HPV16+) in a phase II trial (14, 15). Sufficient sera from 23 of 27 AGIN patients were available for evaluation. Proliferation assays. The proliferation assays are described in refs. 12, 14.