assessment of humoral immune response of a cytomegalovirus dna-vaccine candidate in balb/c mice

Authors

r vahabpour pasteur institute of iran

mr aghasadeghi pasteur institute of iran

f goudarzifar pasteur institute of iran

h keyvani pasteur institute of iran

abstract

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.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

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 evaluated. Methods: The truncated gB coding sequence was synthesized and cloned in pEGFPN1 eukary...

full text

Induction of Specific Humoral Immune Response in Mice against a Pseudomonas aeruginosa Chimeric PilQ/PilA Protein

Background: Pseudomonas aeruginosa, an opportunistic pathogen, is a common cause of healthcare-associated infections in immunocompromised individuals. The rapid emergence of multidrug-resistant strains has made P. aeruginosa infections progressively difficult to treat. In this study we evaluated the effect of a chimeric protein containing a P. aeruginosa PilQ fragment and the PilA disulfide loo...

full text

Xanthan gum enhances humoral immune response elicited by a DNA vaccine against leptospirosis in mice

Background Traditional vaccines (killed or inactivated) played vital roles in controlling and eradicating infectious diseases for a long time. Antigen-specific T cell response can be induced when mice are intramuscularly inoculated with naked plasmid DNA. Therefore, DNA vaccines were evaluated in many studies, demonstrating its safety, stability and easy production. Furthermore, it has been rep...

full text

Induction of humoral immune responses and inhibition of metastasis in mice by a VEGF peptide-based vaccine

Objective(s): Blocking of vascular endothelial growth factor (VEGF) plays a pivotal role in inhibition of metastasis and is a target for development of anti-angiogenic agents. In this study, a peptide-based vaccine was designed and its potential for induction of humoral immune responses, generation of neutralizing antibodies, inhibition of tumor growth and metastasis w...

full text

CFP10: mFcγ2 as a novel tuberculosis vaccine candidate increases immune response in mouse

Objective(s): Despite treatment with antibiotics and vaccination with BCG, tuberculosis (TB) is still considered as one of the most important public health problems in the world. Therefore, designing and producing a more effective vaccine against TB seems urgently. In this study, immunogenicity of a fusion protein which consisting or comprising CFP-10 from Mycobacterium tuberculosis and the Fc-...

full text

The Effect of Cationic Liposomes Encapsulating pcDNA3.1+PA Plasmids on Humoral Immune Response in Mice

Background: DNA vaccines are third generation vaccines which have made promises to combat infectious diseases. Cationic liposomes are used as effective delivery systems for DNA vaccines to generate stronger immunity. Objective: Encapsulation of pcDNA3.1+PA plasmid, encoding protective antigen (PA) of Bacillus anthracis (B. anthracis) into cationic liposomes, and evaluation of its effect on spec...

full text

My Resources

Save resource for easier access later


Journal title:
vaccine research

جلد ۲، شماره ۳، صفحات ۳۳-۳۷

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023