• Title/Summary/Keyword: Human Papillomavirus 16

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Engineering of Recombinant Human Papillomavirus 16 L1 Protein for Incorporation with para-Azido-L-Phenylalanine

  • Jinhyeon Kim;Ki Jun Jeong;Geun-Joong Kim;Jong-il Choi
    • Journal of Microbiology and Biotechnology
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    • v.34 no.9
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    • pp.1926-1932
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    • 2024
  • Human papillomavirus (HPV) L1 capsid protein were produced in several host systems, but few studies have focused on enhancing the properties of the L1 protein. In this study, we aimed to produce recombinant Human papillomavirus (HPV) L1 capsid protein containing para-azido-L-phenylalanine (pAzF) in Escherichia coli. First, we expressed the maltose-binding protein (MBP)-fused HPV16 L1, and 5 residues in HPV16 L1 protein were selected by the in silico modeling for amber codon substitution. Among the variants of the five locations, we identified a candidate that exhibited significant differences in expression with and without pAzF via genetic code expansion (GCE). The expressed recombinant MBP-HPV16L1 protein was confirmed for incorporation of pAzF and the formation of VLPs was tested in vitro.

Establishment and Characterization of Immortalized Human Dermal Papilla Cells Expressing Human Papillomavirus 16 E6/E7

  • Seonhwa Kim;Kyeong-Bae Jeon;Hyo-Min Park;Jinju Kim;Chae-Min Lim;Do-Young Yoon
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.506-515
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    • 2024
  • Primary human dermal papilla cells (HDPCs) are often preferred in studies on hair growth and regeneration. However, primary HDPCs are limited by their reduced proliferative capacity, decreased hair induction potential, and extended doubling times at higher passages. To overcome these limitations, pTARGET vectors containing human papillomavirus16 (HPV16) E6/E7 oncogenes were transfected into HDPCs and selected using G-148 to generate immortalized cells here. HPV16 E6/E7 oncogenes were efficiently transfected into primary HDPCs. Immortalized HDPC showed higher proliferative activity than primary HDPC, confirming an increased proliferation rate. Expression of p53 and pRb proteins was downregulated by E6 and E7, respectively. E6/E7 expressing HDPC cells revealed that cyclin-dependent kinase (CDK) inhibitor p21 expression was decreased, while cell cycle-related genes and proteins (CDK2 and cyclin E) and E2F family genes were upregulated. Immortalized HDPCs maintained their responsiveness to Wnt/β-catenin pathway and hair follicle formation capability, as indicated by their aggregative properties and stemness. E6/E7 immortalized HDPCs may facilitate in vitro hair growth and regeneration studies.

Human Papillomavirus Type 16/18 Oncoproteins: Potential Therapeutic Targets in Non-smoking Associated Lung Cancer

  • Zhang, Er-Ying;Tang, Xu-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5363-5369
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    • 2012
  • High-risk human papillomavirus (HPV) especially HPV-16 and HPV-18 types are speculated to be important risk factors in non-smoking associated lung cancer in Asia. Increasing evidence has demonstrated that HPV oncoproteins may contribute to lung tumorigenesis and cell transformation. Importantly, HPV 16/18 E6 and E7 oncoproteins can mediate expression of multiple target genes and proteins, such as p53/pRb, VEGF, HIF-$1{\alpha}$, cIAP-2, and hTERT, and contribute to cell proliferation, angiogenesis and cell immortalization through different signaling pathways in lung cancer. This article provides an overview of experiment data on HPV-associated lung cancer, describes the main targets on which HPV E6/E7 oncoproteins act, and further discusses the potential signaling pathways in which HPV E6/E7 oncoproteins are involved. In addition, we also raise questions regarding existing problems with the study of HPV-associated lung cancer.

Production and Prophylactic Efficacy Study of Human Papillomavirus-like Particle Expressing HPV16 L1 Capsid Protein

  • Park, Jie-Yun;Pyo, Hyun-Mi;Yoon, Sun-Woo;Baek, Sun-Young;Park, Sue-nie;Kim, Chul-Joong;Haryoung Poo
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.313-318
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    • 2002
  • To perform the prophylactic study of a vaccine derived from human papillomavirus (HPV) using Balb/c mice, we produced virus like particles consisting of HPV capsid protein L1 which has been reported to induce significant humoral and cellular immunity using various animal model systems. In order to produce HPV16 VLPs, the cDNA of L1 capsid protein in HPV type 16, obtained by polymerase chain reaction, was inserted into yeast expression vector, YEG$\alpha$-HIR525 under the control of GAL10 promoter. The transformation of YEG$\alpha$-HPV16 L1 was performed into the yeast Saccharomyces cerevisiae Y2805 by the lithium acetate method and the yeast clone expressing the highest level of L1 capsid protein of human papillomavirus type 16 was selected by Western blot analysis using anti-HPV16 L1 antibody. The purification of HPV16 VLP has been performed by the ultracentrifugation and gel-filtration methods. To validate the vaccine efficacy of the purified HPV16 VLPs and investigate the properties of HPV16 VLPs to induce humoral immunity, ELISA assay was performed. A significantly increased production of anti-HPV16 VLP antibodies was observed in sera from immunized mice. The neutralization activity of antibodies in the sera from the vaccinated mice was demonstrated by a rapid and simple assay to detect hemagglutihation inhibition activity.

Cervical Pathology in High-Risk Human Papillomavirus-Positive, Cytologically Normal Women

  • Supho, Buasorn;Supoken, Amornrat;Kleebkaew, Pilaiwan;Kietpeerakool, Chumnan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7977-7980
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    • 2014
  • This study was undertaken to evaluate the prevalence of significant cervical pathology among women who are high-risk human papillomavirus (HR-HPV)-positive/cytology negative, the most common combination of positive co-tests. The records of 244 women HR-HPV-positive/cytology-negative who had undergone colposcopy at Srinagarind Hospital, Khon Kaen University during January 2010 and April 2014 were reviewed. Mean age was 46.4 years. Of these 224 women, 75 were positive for HPV types 16/18 (33.5%) and 123 were positive for non-16/18 types (54.9%). HR-HPV was not genotyped in the remaining 26 women (11.6%). Prevalence of significant lesions for the entire cohort was 2.4%, and 2.6% and 3.3%, respectively, for those with HPV 16/18 and other oncogenic HPV types. One woman with HPV 16/18 (1.3%) had invasive cervical cancer. Multiparous women were more likely to be infected with HPV 16/18 compared to nulliparous women (36.3% versus 17.6%, respectively). In conclusion, the prevalence of significant cervical lesion among our study population was 2.4%. Multiparous women were more likely to be infected with HPV 16/18 compared to nulliparous women.

Establishment of Transgenic Mouse with the E6 and E7 Genes of Human Papillomavirus Type 16 (인간 Papillomavirus의 E6, E7 유전자를 이용한 Transgenic Mouse의 확립)

  • Hwang, Yong-Il;Lee, Seung-Cheol;Kim, Hyun-Su
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.115-120
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    • 1996
  • Human papillomavirus (HPV), especially type 16 and 18, has been closely associated with carcinomas and uterine cevical cancer, recently. From in vitro assays, E6 and E7 genes of HPV16 are closely linked with transformation of cell lines of rodent fibroplasts. However, the transforming activity of E6 and E7 genes of HPV type 16 in vivo has not been fully elucidated. For explaining this mechanism, we prepared a expression system with the promoter of mouse mammary tumorvirus long terminal repeat and E6E7's open reading frames. This expression system was introduced in rodent cell lines, No. 7, 3Y1 and shown normal transforming abilities. And, we produced transgenic mice with E6, E7 expression system. These transgenic mice were confirmed from Southern blot analysis. One male of them was observed enlargement of the testis after 5 months postdelivery.

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Human Papilloma Virus Attributable Head and Neck Cancer in the Sudan Assessed by p16INK4A Immunostaining

  • Ahmed, Hussain Gadelkarim;Mustafa, Saadalnour Abusail;Warille, Eyman
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6083-6086
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    • 2012
  • Background: The aim of this study was to screen for human papillomavirus (HPV) infections in head and neck squamous cell carcinomas (HNSCCs) using P16 immunostaining. Materials and Methods: A retrospective study was performed on 150 samples from patients diagnosed with HNSCCs. HPV status was determined using $p16^{INK4A}$. Results: 31 of the 150 (20.7%) HNSCCs were HPV positive. Conclusions: A large proportion of HNSCCs in Sudan are associated with HPV infection. The fact that the prevalence of HPV is high among Sudanese patients with head and neck cancers (HNC) has obvious implications for vaccine therapy.

Prevalence of Human Papillomavirus in Women from Saudi Arabia

  • Turki, Rola;Sait, Khalid;Anfinan, Nisreen;Sohrab, Sayed Sartaj;Abuzenadah, Adel Mohammed
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3177-3181
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    • 2013
  • Background: Human papillomavirus (HPV) infection is the main causes of cervical cancer in women worldwide. The goal of the present study was to determine the prevalence and distribution of HPV genotypes in women from Saudi Arabia. Recently, several HPV detection methods have been developed, each with different sensitivities and specificities. Methods: In this study, total forty cervical samples were subjected to polymerase chain reaction and hybridization to BioFilmChip microarray assessment. Results: Human papillomavirus (HPV) infections were found in 43% of the specimens. The most prevalent genotypes were HPV 16 (30%) HPV 18 (8.0%) followed by type HPV 45, occurring at 5.0%. Conclusion: Our finding showed the HPV infection and prevalence is increasing at alarming rate in women of Saudi Arabia. There was no low risk infection detected in the tested samples. The BioFilmChip microarray detection system is highly accurate and suitable for detection of single and multiple infections, allowing rapid detection with less time-consumption and easier performance as compared with other methods.

HPV DNA TYPING OF ADULT-ONSET LARYNGEAL PAPILLOMATOSIS (성인의 후두유두종에 대한 Human Papillomavirus의 DNA Typing)

  • 장문수;임영묵;나기상;유장렬;박찬일
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1991.06a
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    • pp.19-19
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    • 1991
  • 성인의 후두유두종은 Human Papillonavirus에 의해 발생하는 것으로 알려져 있다. 저자들은 1985년 4월부터 1989년 12월까지 충남대학교병원 이비인후과에서 후두유두종으로 수술받은 19예중 보관된 Paraffin block의 이용이 가능하였던 14예에서 Human Papillomavirus type 6, 11, 16 및 18에 대해 In situ hybridization 방법으로 DNA typing을 시행하여 다음과 같은 결과를 얻었다. 14예중 10예에서 type 6에 양성을 보였고 type 6에 양성을 보인 10예중 6예에서 type 11에 대해서도 양성을 보였다. Type 11 단독으로 양성을 보인 예는 없었다. 또한, type 16 및 18에 대해서는 모두 음성을 보였다. 이상의 결과로 성인의 후두유두종은 Human Papillomavirus type 6 및 11과 밀접한 관계가 있을 것으로 사료된다.

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Whole Genome Analysis of Human Papillomavirus Type 16 Multiple Infection in Cervical Cancer Patients

  • Chansaenroj, Jira;Theamboonlers, Apiradee;Junyangdikul, Pairoj;Swangvaree, Sukumarn;Karalak, Anant;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.599-606
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    • 2012
  • The characterization of the whole genome of human papillomavirus type 16 (HPV16) from cervical cancer specimens with multiple infections in comparison with single infection samples as the oncogenic potential of the virus may differ. Cervical carcinoma specimens positive for HPV16 by PCR and INNO-LiPA were randomly selected for whole genome characterization. Two HPV16 single infection and six HPV16 multiple infection specimens were subjected to whole genome analysis by using conserved primers and subsequent sequencing. All HPV16 whole genomes from single infection samples clustered in the European (E) lineage while all multiple infection specimens belonged to the non-European lineage. The variations in nucleotide sequences in E6, E7, E2, L1 and Long control region (LCR) were evaluated. In the E6 region, amino acid changes at L83V were related to increased cancer progression. An amino acid variation N29S within the E7 oncoprotein significantly associated with severity of lesion was also discovered. In all three domains of the E2 gene non synonymous mutations were found. The L1 region showed various mutations which may be related to conformation changes of viral epitopes. Some transcription factor binding sites in the LCR region correlated to virulence were shown on GRE/1, TEF-1, YY14 and Oct-1. HPV16 European variant prone to single infection may harbor a major variation at L83V which significantly increases the risk for developing cervical carcinoma. HPV16 non-European variants prone to multiple infections may require many polymorphisms to enhance the risk of cervical cancer development.