Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice
Introduction: Glycoprotein B (gB) is the major antigen for induction of humoral responses against human cytomegalovirus (HCMV) making it an attractive candidate for immune prophylaxis. In the present study, the humoral immune response of BALB/c mice to a truncated HCMV gB protein fused with GFP was...
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| Format: | Article |
| Language: | English |
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Pasteur Institute of Iran
2015-05-01
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| Series: | Vaccine Research |
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| Online Access: | http://vacres.pasteur.ac.ir/article-1-61-en.html |
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| author | R Vahabpour MR Aghasadeghi F Goudarzifar H Keyvani A Ataei- Pirkooh SH Monavari M Eslami |
| author_facet | R Vahabpour MR Aghasadeghi F Goudarzifar H Keyvani A Ataei- Pirkooh SH Monavari M Eslami |
| author_sort | R Vahabpour |
| collection | DOAJ |
| description | Introduction: Glycoprotein B (gB) is the major antigen for induction of humoral responses against human cytomegalovirus (HCMV) making it an attractive candidate for immune prophylaxis. In the present study, the humoral immune response of BALB/c mice to a truncated HCMV gB protein fused with GFP was evaluated. Methods: The truncated gB coding sequence was synthesized and cloned in pEGFPN1 eukaryotic expression vector and expressed in HEK 293T cell line. After optimization, expression of the recombinant truncated HCMV gB was verified using HRP-conjugated polyclonal antibody specific for HCMV gB.The level of humoral immune responses was assessed in mice using DNA/DNA, peptide/peptide, and DNA/ peptide (prime-boost) immunization strategies. Results: Cloning of the truncated gB coding sequence in the pEGFPN1 was verified by restriction enzyme analysis and sequencing. After optimizing the transfection procedure the number of the GFP positive cells reached 32%. Western blot analysis confirmed the in vitro expression of the truncated HCMV gB protein with an apparent molecular weight of approximately 70 kDa. In vivo prime-boost immunization using HCMV gB DNA/peptide regimen showed significantly higher humoral immune responses compared to the control groups. Conclusion: This study demonstrated that the pEGFPN1 eukaryotic expression vector could be used to optimize and evaluate the expression of this truncated protein.The results also showed that the DNA/peptide vaccination could induce a significant antibody response in animal model. |
| first_indexed | 2024-12-20T02:03:41Z |
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| id | doaj.art-5740ef0890894fbb8699cfaff1956e3a |
| institution | Directory Open Access Journal |
| issn | 2383-2819 2423-4923 |
| language | English |
| last_indexed | 2024-12-20T02:03:41Z |
| publishDate | 2015-05-01 |
| publisher | Pasteur Institute of Iran |
| record_format | Article |
| series | Vaccine Research |
| spelling | doaj.art-5740ef0890894fbb8699cfaff1956e3a2022-12-21T19:57:15ZengPasteur Institute of IranVaccine Research2383-28192423-49232015-05-01213337Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c miceR Vahabpour0MR Aghasadeghi1F Goudarzifar2H Keyvani3A Ataei- Pirkooh4SH Monavari5M Eslami6 Pasteur Institute of Iran Pasteur Institute of Iran Pasteur Institute of Iran Pasteur Institute of Iran Pasteur Institute of Iran Pasteur Institute of Iran Pasteur Institute of Iran Introduction: Glycoprotein B (gB) is the major antigen for induction of humoral responses against human cytomegalovirus (HCMV) making it an attractive candidate for immune prophylaxis. In the present study, the humoral immune response of BALB/c mice to a truncated HCMV gB protein fused with GFP was evaluated. Methods: The truncated gB coding sequence was synthesized and cloned in pEGFPN1 eukaryotic expression vector and expressed in HEK 293T cell line. After optimization, expression of the recombinant truncated HCMV gB was verified using HRP-conjugated polyclonal antibody specific for HCMV gB.The level of humoral immune responses was assessed in mice using DNA/DNA, peptide/peptide, and DNA/ peptide (prime-boost) immunization strategies. Results: Cloning of the truncated gB coding sequence in the pEGFPN1 was verified by restriction enzyme analysis and sequencing. After optimizing the transfection procedure the number of the GFP positive cells reached 32%. Western blot analysis confirmed the in vitro expression of the truncated HCMV gB protein with an apparent molecular weight of approximately 70 kDa. In vivo prime-boost immunization using HCMV gB DNA/peptide regimen showed significantly higher humoral immune responses compared to the control groups. Conclusion: This study demonstrated that the pEGFPN1 eukaryotic expression vector could be used to optimize and evaluate the expression of this truncated protein.The results also showed that the DNA/peptide vaccination could induce a significant antibody response in animal model.http://vacres.pasteur.ac.ir/article-1-61-en.htmlhuman cytomegalovirusglycoprotein bdna-based vaccine. |
| spellingShingle | R Vahabpour MR Aghasadeghi F Goudarzifar H Keyvani A Ataei- Pirkooh SH Monavari M Eslami Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice Vaccine Research human cytomegalovirus glycoprotein b dna-based vaccine. |
| title | Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice |
| title_full | Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice |
| title_fullStr | Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice |
| title_full_unstemmed | Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice |
| title_short | Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice |
| title_sort | assessment of humoral immune response of a cytomegalovirus dna vaccine candidate in balb c mice |
| topic | human cytomegalovirus glycoprotein b dna-based vaccine. |
| url | http://vacres.pasteur.ac.ir/article-1-61-en.html |
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