Human Papillomavirus Type 16 Exists in Bacteria Isolated from Cervical Cancer Biopsies

2009 ◽  
Vol 37 (4) ◽  
pp. 1065-1074 ◽  
Author(s):  
Zhenghai Ma ◽  
Lihong Liu ◽  
Fuchun Zhang ◽  
Meng Yu ◽  
Kai Wang ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
Reverse Transcription Pcr ◽  
Bacterial Rna ◽  
L1 Protein

This study investigated the association between infectious microbes and persistent infection with human papillomavirus type 16 (HPV-16) in cervical cancer. Bacterial strains (identified as Enterococcus, Staphylococcus, Bacillus and Corynebacterium, based on their partial 16S rDNA sequence) were HPV-16 positive from 12 out of 14 cervical cancer biopsies. Total DNA was isolated from the four bacterial strains, and HPV-16 genes and genome were detected using polymerase chain reaction (PCR) and Southern blotting. RNA transcripts for HPV-16 E6 and L1 genes were detected in total bacterial RNA samples using reverse transcription-PCR, and HPV-16 L1 protein expression was detected in bacterial cells by Western blotting and immunocolloidal gold electron microscopy. The presence of virus particles in bacterial cells was demonstrated by transmission electron microscopy. The results suggest that bacteria carrying HPV-16 could provide a potential explanation for how infectious microbes contribute to the progression from HPV-16 infection to cervical cancer.

scholarly journals Multiple ASF/SF2 Sites in the Human Papillomavirus Type 16 (HPV-16) E4-Coding Region Promote Splicing to the Most Commonly Used 3′-Splice Site on the HPV-16 Genome

2010 ◽  
Vol 84 (16) ◽  
pp. 8219-8230 ◽  
Author(s):  
Monika Somberg ◽  
Stefan Schwartz
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Splice Site ◽  
Binding Sites ◽  
Mrna Levels ◽  
Coding Region ◽  
Cervical Lesions ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
E6 And E7

ABSTRACT Our results presented here demonstrate that the most abundant human papillomavirus type 16 (HPV-16) mRNAs expressing the viral oncogenes E6 and E7 are regulated by cellular ASF/SF2, itself defined as a proto-oncogene and overexpressed in cervical cancer cells. We show that the most frequently used 3′-splice site on the HPV-16 genome, site SA3358, which is used to produce primarily E4, E6, and E7 mRNAs, is regulated by ASF/SF2. Splice site SA3358 is immediately followed by 15 potential binding sites for the splicing factor ASF/SF2. Recombinant ASF/SF2 binds to the cluster of ASF/SF2 sites. Mutational inactivation of all 15 sites abolished splicing to SA3358 and redirected splicing to the downstream-located, late 3′-splice site SA5639. Overexpression of a mutant ASF/SF2 protein that lacks the RS domain, also totally inhibited the usage of SA3358 and redirected splicing to the late 3′-splice site SA5639. The 15 ASF/SF2 binding sites could be replaced by an ASF/SF2-dependent, HIV-1-derived splicing enhancer named GAR. This enhancer was also inhibited by the mutant ASF/SF2 protein that lacks the RS domain. Finally, silencer RNA (siRNA)-mediated knockdown of ASF/SF2 caused a reduction in spliced HPV-16 mRNA levels. Taken together, our results demonstrate that the major HPV-16 3′-splice site SA3358 is dependent on ASF/SF2. SA3358 is used by the most abundantly expressed HPV-16 mRNAs, including those encoding E6 and E7. High levels of ASF/SF2 may therefore be a requirement for progression to cervical cancer. This is supported by our earlier findings that ASF/SF2 is overexpressed in high-grade cervical lesions and cervical cancer.

scholarly journals Cloning and expression of Human Papilloma virus type 16 L1 capsid protein in bacteria

2019 ◽  
Vol 10 (2) ◽  
pp. 82-89
Author(s):  
Silvia Tri Widyaningtyas ◽  
Sofy Meilany ◽  
Budiman Bela
Keyword(s):  
Escherichia Coli ◽  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Recombinant Protein ◽  
Capsid Protein ◽  
Recombinant Plasmid ◽  
Health Science ◽  
Science Journal ◽  
Hpv 16 ◽  
L1 Protein

Latar belakang: Secara alamiah protein kapsid L1 Human Papillomavirus (HPV) tipe 16 dapat mengalami auto assembly untuk membentuk Viral like particle (VLP). Terkait dengan penelitian vaksin HPV, VLP dapat digunakan untuk berbagai keperluan seperti vaksin, pseudovirion atau SpyTag-Spycatcher. Penelitian ini ditujukan untuk mendapatkan plasmid rekombinan yang digunakan untuk produksi protein L1 HPV 16. Metode: Gen penyandi protein L1 HPV 16 diklona ke dalam vector pQE80L, suatu plasmid yang mengandung sistem ekspresi untuk prokariota. DNA penyandi HPV 16 L1 disisipkan pada situs restriksi BamHI dan Hind III plasmid pQE80L. Plasmid rekombinan yang mengandung gen L1 HPV 16dikonfirmasi menggunakan PCR dan analisis enzim restriksi. Lebih lanjut untuk memastikan bahwa gen rekombinan L1 HPV 16 dapat diekspresikan dalam prokariota, plasmid rekombinan ditransformasikan ke bakteri Escherichia coli BL21 (DE3). Bakteri diinduksi dengan Isopropyl β-D-1-thiogalactopyranoside (IPTG) dengan berbagai konsentrasi dan berbagai waktu inkubasi. Hasil: protein rekombinan L1, berat 56 kDa, telah berhasil diekspresikan dalam sistem prokariota. Protein rekombinan L1 dapat dimurnikan menggunakan TalonR dalam kondisi denaturasi. Kesimpulan: gen L1 HPV 16 telah dikloning ke dalam pQE80L dan berhasil diekspresikan dalam sistem prokariota. (Health Science Journal of Indonesia 2019;10(2):82-9) Kata kunci: L1, HPV 16, cervical cancer   Abstract Background: Naturally Human Papillomavirus (HPV) type 16 L1 capsid protein can auto assemble to form Viral like particles (VLP). Concerning to vaccine development for HPV, VLP can be used for a variety of needs such as a vaccine, pseudovirion or SpyTag-Spycatcher. In this study, to obtain a vector expression that can be used in the production of HPV L1 protein, we cloned gene coding HPV 16 L1 protein into pQE80L a plasmid contains an expression system for prokaryote. Methods: The DNA coding HPV 16 L1 was inserted at BamHI and Hind III restriction sites of pQE80L plasmid. The recombinant plasmid containing the HPV L1 gene was confirmed using PCR colony and enzyme restriction. Further to ensure the recombinant HPV 16 L1 gene could be expressed in a prokaryote, the recombinant plasmid was transformed into bacteria Escherichia coli BL21 (DE3). The bacteria were induced with IPTG with various concentrations and various incubation time. Result: L1 recombinant protein, 56 kDa in weight, has successfully been expressed in prokaryote system. L1 recombinant protein can be purified using TalonR under denaturing conditions. Conclusion: L1 HPV 16 gene has been cloned into pQE80L and successfully expressed in prokaryote system. (Health Science Journal of Indonesia 2019;10(2):82-9) Keywords: L1, HPV 16, cervical cancer

The interaction between steroid hormones, human papillomavirus type 16, E6 oncogene expression, and cervical cancer

2003 ◽  
Vol 13 (6) ◽  
pp. 834-842 ◽  
Author(s):  
M. Moodley ◽  
S. Sewart ◽  
C. S. Herrington ◽  
R. Chetty ◽  
R. Pegoraro ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Steroid Hormones ◽  
Housekeeping Gene ◽  
Hpv 16 ◽  
Tissue Samples ◽  
Hpv Dna ◽  
Human Papillomavirus Type 16 ◽  
Transforming Proteins

Various risk factors have been implicated in the causation of cervical cancer including human papillomavirus (HPV), the early genes (E6 and E7) of which encode the main transforming proteins. Studies have suggested that steroid hormones may enhance the expression of these genes leading to loss of p53 gene-mediated cell apoptosis. A total of 120 cervical tissue samples were obtained from patients with proven cervical cancer. Patients who used depo-medroxyprogesterone acetate steroid contraception were recruited as part of the steroid arm. Only HPV DNA type 16 samples were used for the study. Controls included three cell lines (CaSki, SiHa, & C33A) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal housekeeping gene. Of 120 patients, there were 111 patients with HPV type 16 identified. Of this number, RNA was present in 63 samples. There were 30 women (30/63) who used steroid contraception. In relation to patients who used contraception, HPV 16 E6 gene expression was present in 79% (n = 23) and 88% (n = 30) of steroid users compared to nonusers, respectively. In total there were 25 patients (40%) with expression of the HPV 16 E6*I gene and 30 patients with expression of the E6*II gene. There were 57% of steroid users (n = 17) who had expression of the E6*I/E6*II gene, compared to 52% (n = 17) of nonusers (P = 0.800). From a molecular level, this study does not confirm the role of injectable progesterones in cervical carcinogenesis.

scholarly journals Production of Human Papillomavirus Type 16 L1 Virus-Like Particles by Recombinant Lactobacillus casei Cells

2006 ◽  
Vol 72 (1) ◽  
pp. 745-752 ◽  
Author(s):  
Karina Araujo Aires ◽  
Aurora Marques Cianciarullo ◽  
Sylvia Mendes Carneiro ◽  
Luisa Lina Villa ◽  
Enrique Boccardo ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Lactobacillus Casei ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hpv 16 ◽  
Virus Like Particles ◽  
Human Papillomavirus Type 16 ◽  
Common Causes ◽  
Electron Microscopy Analysis ◽  
Microscopy Analysis ◽  
L1 Protein

ABSTRACT Infections with human papillomavirus type 16 (HPV-16) are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, the most promising vaccine against HPV-16 infection is based on the L1 major capsid protein, which self-assembles in virus-like particles (VLPs). In this work, we used a lactose-inducible system based on the Lactobacillus casei lactose operon promoter (plac) for expression of the HPV-16 L1 protein in L. casei. Expression was confirmed by Western blotting, and an electron microscopy analysis of L. casei expressing L1 showed that the protein was able to self-assemble into VLPs intracellularly. The presence of conformational epitopes on the L. casei-produced VLPs was confirmed by immunofluorescence using the anti-HPV-16 VLP conformational antibody H16.V5. Moreover, sera from mice that were subcutaneously immunized with L. casei expressing L1 reacted with Spodoptera frugiperda-produced HPV-16 L1 VLPs, as determined by an enzyme-linked immunosorbent assay. The production of L1 VLPs by Lactobacillus opens the possibility for development of new live mucosal prophylactic vaccines.

scholarly journals Cytotoxic T-Lymphocyte Responses to Human Papillomavirus Type 16 E5 and E7 Proteins and HLA-A*0201-Restricted T-Cell Peptides in Cervical Cancer Patients

2007 ◽  
Vol 81 (6) ◽  
pp. 2869-2879 ◽  
Author(s):  
Dai-Wei Liu ◽  
Yuh-Cheng Yang ◽  
Ho-Fan Lin ◽  
Mei-Fang Lin ◽  
Ya-Wen Cheng ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
T Cell ◽  
Cancer Patients ◽  
T Lymphocyte ◽  
Cytotoxic T Lymphocyte ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
E5 Protein ◽  
E7 Proteins

ABSTRACT Previously, we found that human papillomavirus type 16 (HPV-16) E5 protein is a tumor rejection antigen and can induce cytotoxic T-lymphocyte (CTL) activity. Therefore, in this study, human leukocyte antigen A*0201 (HLA-A*0201)-restricted human CTL epitopes of HPV-16 E5 protein were identified using a bioinformatics approach, and the abilities of these predicted peptides to induce an immune response in HLA-A*0201 transgenic mice were confirmed by assaying E5-specific CTLs and in vitro-generated CTLs from normal peripheral blood T lymphocytes of HLA-A2-positive human donors. Second, the CTL responses to HLA-A*0201 CTL epitopes (E5 63-71 and E7 11-20) were examined in HPV-16-infected patients with HLA-A2. Third, the effect of HLA-A-type alleles on CTL activities in response to the entire E5 and E7 proteins was examined in cervical cancer patients. E5 and E7 peptides (but not the whole proteins) stimulated E5- and E7-specific CTL recall responses in HPV-16- and HLA-A2-positive cervical cancer patients, and HPV-16 E5 and E7 proteins stimulated naïve T cells in HPV-16-negative cervical cancer patients with HLA-A11 and -A24 haplotypes. In summary, this is the first demonstration that E5 63-71 is an HLA-A*0201-restricted T-cell epitope of HPV-16 E5.

scholarly journals Influence of Codon Bias on Heterologous Production of Human Papillomavirus Type 16 Major Structural Protein L1 in Yeast

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Milda Norkiene ◽  
Alma Gedvilaite
Keyword(s):  
Gene Expression ◽  
Human Papillomavirus ◽  
Codon Bias ◽  
Heterologous Gene Expression ◽  
Heterologous Production ◽  
Structural Protein ◽  
Hpv 16 ◽  
Major Structural Protein ◽  
Human Papillomavirus Type 16 ◽  
L1 Protein

Heterologous gene expression is dependent on multistep processes involving regulation at the level of transcription, mRNA turnover, protein translation, and posttranslational modifications. Codon bias has a significant influence on protein yields. However, sometimes it is not clear which parameter causes observed differences in heterologous gene expression as codon adaptation typically optimizes many sequence properties at once. In the current study, we evaluated the influence of codon bias on heterologous production of human papillomavirus type 16 (HPV-16) major structural protein L1 in yeast by expressing five variants of codon-modified open reading frames (OFRs) encoding HPV-16 L1 protein. Our results showed that despite the high toleration of various codons used throughout the length of the sequence of heterologously expressed genes in transformed yeast, there was a significant positive correlation between the gene's expression level and the degree of its codon bias towards the favorable codon usage. The HPV-16 L1 protein expression in yeast can be optimized by adjusting codon composition towards the most preferred codon adaptation, and this effect most probably is dependent on the improved translational elongation.

Genetic variability in the major capsid L1 protein of human papillomavirus type 16 (HPV-16) and 18 (HPV-18)

2011 ◽  
Vol 11 (8) ◽  
pp. 2119-2124 ◽  
Author(s):  
E. Frati ◽  
S. Bianchi ◽  
D. Colzani ◽  
A. Zappa ◽  
G. Orlando ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Genetic Variability ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
L1 Protein ◽  
Hpv 18

scholarly journals Concordance of Human Papillomavirus Type 16 and 18 in Cervical and Oral Specimen of Cervical Cancer Patients

2019 ◽  
pp. 70
Author(s):  
Willy Akbar ◽  
Syahrul Rauf ◽  
Deviana S. Riu ◽  
St. Maisuri T. Chalid
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Oral Cavity ◽  
Cancer Patients ◽  
Oral Fluid ◽  
Cross Sectional Study ◽  
Cross Sectional ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
Oral Hpv

Abstract Objective : To determine the conformity of HPV type 16 and 18 in cervical and oral/buccal specimens from cervical cancer patients. Methods :A cross-sectional study was conducted in March - September 2016 at several hospitals in Makassar. HPV 16 and 18 genotyping in cervical and oral fluid of 77 patients with cervical cancer performed with PCR method. Results : The prevalence of HPV type 18 infection both in the cervical and the oral fluid was higher than HPV type 16 [9(47.4%) vs 5(26.3%)]. The aggreement of HPV type 18 infection (r=0.328;p=0.000) in the cervical-oral sites was higher than HPV type 16 (r=0.194;p=0.042). Conclusion : HPV type 16 and 18 could infect both cervix and oral cavity although type-specific concordance is low. Keywords :Human papillomavirus,servix, oral cavity   Abstrak Tujuan: Mengetahui tingkat kesesuaian hasil pemeriksaan HPV tipe 16 dan 18 antara spesimen serviks dan oral/buccal pada penderita kanker serviks. Metode: Penelitian cross sectional ini dilakukan pada Maret – September 2016 pada beberapa rumah sakit di Makassar. Pemeriksaan HPV 16 dan 18 pada cairan serviks dan oral dari 77 orang penderita kanker serviks menggunakan teknik PCR. Hasil: Prevalensi infeksi bersama pada serviks dan oral HPV tipe 18 lebih tinggi dibandingkan HPV tipe 16 [9(47,4%) vs 5(26,3%)]. Tingkat kesesuaian antara HPV tipe 18 (r=0,328;p=0,000) pada serviks dan oral lebih tinggi dibandingkan tipe 16 (r=0,194;p=0,042). Kesimpulan: HPV tipe 16 dan 18 dapat menginfeksi serviks dan oral meskipun tingkat kesesuaian kedua tipe ini lemah. Kata kunci : Human papillomavirus, serviks, kavum oral

scholarly journals Human Papillomavirus Types 16, 31, and 58 Use Different Endocytosis Pathways To Enter Cells

2003 ◽  
Vol 77 (6) ◽  
pp. 3846-3850 ◽  
Author(s):  
Latifa Bousarghin ◽  
Antoine Touzé ◽  
Pierre-Yves Sizaret ◽  
Pierre Coursaget
Keyword(s):  
Electron Microscopy ◽  
Human Papillomavirus ◽  
Intracellular Trafficking ◽  
Dominant Negative ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
Virus Like Particle ◽  
Dominant Negative Form ◽  
Negative Form

ABSTRACT The early steps of the intracellular trafficking of human papillomavirus type 16 (HPV-16), -31, and -58 pseudovirions were studied by investigating the effects of drugs acting at defined points of endocytosis pathways on virus-like particle-mediated pseudoinfection by overexpression of a dominant-negative form of the Eps15 protein to inhibit clathrin-mediated endocytosis and by electron microscopy. The results obtained suggested the involvement of clathrin-mediated endocytosis in HPV-16 and HPV-58 entry and caveola-mediated endocytosis in HPV-31 entry.

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