Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18
More than 99% of cervical cancers are associated with human papillomaviruses (HPVs) worldwide. Current HPV vaccines are safe, highly immunogenic, with effective immunity against specific HPV types. However, DNA vaccines are a new appealing platform which can be considered for designing the HPV vacci...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2021-09-01
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| Series: | Iranian Journal of Allergy, Asthma and Immunology |
| Subjects: | |
| Online Access: | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/3092 |
| _version_ | 1818390581716451328 |
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| author | Somayeh Shokri Shahab Mahmoudvand Manoochehr Makvandi Reza Taherkhani Mohammad Rashno Farid Azizi Jalilian Kambiz Ahmadi Angali Mohammad Ali Foroughi |
| author_facet | Somayeh Shokri Shahab Mahmoudvand Manoochehr Makvandi Reza Taherkhani Mohammad Rashno Farid Azizi Jalilian Kambiz Ahmadi Angali Mohammad Ali Foroughi |
| author_sort | Somayeh Shokri |
| collection | DOAJ |
| description | More than 99% of cervical cancers are associated with human papillomaviruses (HPVs) worldwide. Current HPV vaccines are safe, highly immunogenic, with effective immunity against specific HPV types. However, DNA vaccines are a new appealing platform which can be considered for designing the HPV vaccines. This study aimed to construct a recombinant eukaryotic expression plasmid containing L1 of HPV-18, tissue plasminogen activators (tPA), and pan HLA DR-binding epitope (PADRE) genes into the pVAX1 vector.
The L1, tPA, and PADRE genes were amplified in a thermocycler. The polymerase chain reaction (PCR) products were cloned and insertion of the genes was confirmed using colony PCR, restriction enzymes analysis, and sequencing methods. Indirect immunofluorescence, RT-PCR, and western blot assays were applied to identify the target gene in HEK-293 cells. Total IgG and its isotypes in immunized mice were measured by enzyme-linked immunosorbent assay technique.
Western blot analysis showed a protein band of about 67.5 kDa in supernatant and cell lysate of transfected cells. The results of mice immunization with different constructs (group 1: the pVAX-L1, group 2: pVAX-tPA-PADRE-L1, group 3: pVAX1, and group 4: PBS as controls) indicated that the pVAX1-tPA-PADRE-L1 construct induced a significantly higher level of total IgG than pVAX1-L1 (p=0.003).
In conclusion, pVAX1-tPA-PADRE-L1 recombinant plasmid is a highly immunogenic construct and suggests as a promising candidate for vaccine development against HPV type 18 in low-middle-income countries. |
| first_indexed | 2024-12-14T04:59:54Z |
| format | Article |
| id | doaj.art-43755c15c8ca42b096813e1ebd7aeeb6 |
| institution | Directory Open Access Journal |
| issn | 1735-1502 1735-5249 |
| language | English |
| last_indexed | 2024-12-14T04:59:54Z |
| publishDate | 2021-09-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Allergy, Asthma and Immunology |
| spelling | doaj.art-43755c15c8ca42b096813e1ebd7aeeb62022-12-21T23:16:16ZengTehran University of Medical SciencesIranian Journal of Allergy, Asthma and Immunology1735-15021735-52492021-09-0120510.18502/ijaai.v20i5.7403Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18Somayeh Shokri0Shahab Mahmoudvand1Manoochehr Makvandi2Reza Taherkhani3Mohammad Rashno4Farid Azizi Jalilian5Kambiz Ahmadi Angali6Mohammad Ali Foroughi7Infectious and Tropical Disease Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran AND Department of Virology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranInfectious and Tropical Disease Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran AND Department of Virology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranInfectious and Tropical Disease Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran AND Department of Virology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranDepartment of Virology, School of Medicine, Bushehr University of Medical Sciences, Bushehr, IranDepartment of Immunology, Medicine Faculty, Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranDepartment of Virology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, IranDepartment of Biostatistics, School of Health, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IranDepartment of Pathobiology, Section Biotechnology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad IranMore than 99% of cervical cancers are associated with human papillomaviruses (HPVs) worldwide. Current HPV vaccines are safe, highly immunogenic, with effective immunity against specific HPV types. However, DNA vaccines are a new appealing platform which can be considered for designing the HPV vaccines. This study aimed to construct a recombinant eukaryotic expression plasmid containing L1 of HPV-18, tissue plasminogen activators (tPA), and pan HLA DR-binding epitope (PADRE) genes into the pVAX1 vector. The L1, tPA, and PADRE genes were amplified in a thermocycler. The polymerase chain reaction (PCR) products were cloned and insertion of the genes was confirmed using colony PCR, restriction enzymes analysis, and sequencing methods. Indirect immunofluorescence, RT-PCR, and western blot assays were applied to identify the target gene in HEK-293 cells. Total IgG and its isotypes in immunized mice were measured by enzyme-linked immunosorbent assay technique. Western blot analysis showed a protein band of about 67.5 kDa in supernatant and cell lysate of transfected cells. The results of mice immunization with different constructs (group 1: the pVAX-L1, group 2: pVAX-tPA-PADRE-L1, group 3: pVAX1, and group 4: PBS as controls) indicated that the pVAX1-tPA-PADRE-L1 construct induced a significantly higher level of total IgG than pVAX1-L1 (p=0.003). In conclusion, pVAX1-tPA-PADRE-L1 recombinant plasmid is a highly immunogenic construct and suggests as a promising candidate for vaccine development against HPV type 18 in low-middle-income countries.https://ijaai.tums.ac.ir/index.php/ijaai/article/view/3092AlphapapillomavirusesDNA vaccinesIndirect fluorescent antibody techniquePolymerase chain reactionUterine cervical neoplasmsWestern blotting |
| spellingShingle | Somayeh Shokri Shahab Mahmoudvand Manoochehr Makvandi Reza Taherkhani Mohammad Rashno Farid Azizi Jalilian Kambiz Ahmadi Angali Mohammad Ali Foroughi Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18 Iranian Journal of Allergy, Asthma and Immunology Alphapapillomaviruses DNA vaccines Indirect fluorescent antibody technique Polymerase chain reaction Uterine cervical neoplasms Western blotting |
| title | Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18 |
| title_full | Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18 |
| title_fullStr | Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18 |
| title_full_unstemmed | Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18 |
| title_short | Designing of DNA Vaccine Based on a Secretory Form of Major Capsid Protein of Human Papillomavirus Type 18 |
| title_sort | designing of dna vaccine based on a secretory form of major capsid protein of human papillomavirus type 18 |
| topic | Alphapapillomaviruses DNA vaccines Indirect fluorescent antibody technique Polymerase chain reaction Uterine cervical neoplasms Western blotting |
| url | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/3092 |
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