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Reports of Biochemistry and Molecular Biology
rbmb.net
Basic Sciences
http://rbmb.net
1
admin
2322-3480
2322-3480
10.61882/rbmb
en
jalali
1397
10
1
gregorian
2019
1
1
7
2
online
1
fulltext
en
Increasing Cellular Immune Response in Liposomal Formulations of DOTAP Encapsulated by Fusion Protein Hspx, PPE44, And Esxv, as a Potential Tuberculosis Vaccine Candidate
میکروب شناسی
Microbiology
مقالات اصلی
Original Article
<div style="text-align: justify;"><strong><em>Background:</em></strong> Due to the ineffectiveness of the BCG vaccine, especially in adult pulmonary tuberculosis (TB), and variable efficacies against childhood forms of TB, developing an effective TB vaccine is a major priority in controlling this disease. The aim of this study was to evaluate the immunogenicity of a DOTAP liposome formulation containing a fusion protein (FP) containing <em>Mycobacterium tuberculosis</em> HspX, PPE44, and EsxV.<br>
<br>
<strong><em>Methods:</em></strong> The FP was expressed in <em>E. coli</em> BL21 cells and confirmed by SDS-PAGE and Western blots. The FP was then encapsulated in various liposomal formulations. Afterwards, liposomal size, zeta potential, and encapsulation efficiency were evaluated. Mice were subcutaneously vaccinated on days 0, 14, and 28 with liposomes containing the FP. Two weeks after the last injection, IFN-γ, IL-4, IL-17, and IL-12 in spleen cell culture supernatants, and IgG2a, IgG1, and IgG2b titers in sera were measured.<br>
<br>
<strong><em>Results:</em></strong> The FP concentration was 1mg/ml. The encapsulation efficiency of the liposomes varied from 69% in Lip (DOTAP/TDB/CHOL/FP) to 80% in Lip (DOTAP/CHOL/FP). The greatest IFN-γ and IL-12 levels were observed in BCG-primed mice that were boosted with Lip (DOTAP/CHOL/FP). In addition, IL-17 production was significantly greater in all groups than controls except in those that received histidine buffer and FP. The IgG2a/IgG1 ratios were greater in the Lip (DOTAP/TDB/CHOL/FP), Lip (DOTAP/CHOL/FP), Lip (DOTAP/CHOL), and BCG-primed and Lip (DOTAP/CHOL/FP)-boosted groups than in the other groups, indicating a cellular immune response.<br>
<br>
<strong><em>Conclusions:</em></strong> The liposomes containing DOTAP combined with the fusion protein induced a Th1 response. The mice that first received BCG and then Lip (DOTAP/CHOL/FP), produced the most IFN-γ and IL-12, indicating a strong Th1 response.</div>
DOTAP, Vaccines, Fusion Protein, Liposomes, Mycobacterium tuberculosis, Trehalose 6, 6’-Dibehenate.
156
166
http://rbmb.net/browse.php?a_code=A-10-177-1&slc_lang=en&sid=1
Davood
Mansury
mansuryd@varastegan.ac.ir
100319475328460017048
100319475328460017048
No
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. & Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Kiarash
Ghazvini
Ghazvinik@mums.ac.ir
100319475328460017049
100319475328460017049
No
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. & Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Saeid
Amel Jamehdar
ameljs@mums.ac.ir
100319475328460017050
100319475328460017050
No
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran. & Department of Microbiology and Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Ali
Badiee
badieea@mums.ac.ir
100319475328460017051
100319475328460017051
No
Nanotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical Sciences, Mashhad, Iran. & Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Mohsen
Tafaghodi
tafaghodiM@mums.ac.ir
100319475328460017052
100319475328460017052
No
Nanotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical Sciences, Mashhad, Iran. & Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Amin Reza
Nikpoor
nikpoora901@mums.ac.ir
100319475328460017053
100319475328460017053
No
Department of Immunology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Yousef
Amini
AminiY901@mums.ac.ir
100319475328460017054
100319475328460017054
No
Department of Microbiology and Virology, School of Medicine, Zahedan University of Medical Science, Zahedan, Iran.
Mahmoud Reza
Jaafari
JafariMR@mums.ac.ir
100319475328460017055
100319475328460017055
Yes
Nanotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical Sciences, Mashhad, Iran. & Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. & Biotechnology Research Center, Institute of Pharmaceutical Technology, Mashhad University of Medical Sciences, Mashhad, Iran.