caa a22 4500 605502471 CHVBK 20210128100606.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6952-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6952-z APC targeting enhances immunogenicity of a novel multistage Fc-fusion tuberculosis vaccine in mice [Elektronische Daten] [Saman Soleimanpour, Hadi Farsiani, Arman Mosavat, Kiarash Ghazvini, Mohammad Eydgahi, Mojtaba Sankian, Hamid Sadeghian, Zahra Meshkat, Seyed Rezaee] Numerous studies have demonstrated that targeting immunogens to FcγR on antigen-presenting cells (APCs) can selectively uptake and increase cellular immunity in vitro and in vivo. Therefore, the present study was conducted to evaluate immunogenicity of a novel multistage tuberculosis vaccine, a combination of an early and a dormant immunogenic protein, ESAT6 and HspX, fused to Fcγ2a fragment of mouse IgG2a to target all forms of tuberculosis. Codon-optimized genes consisting of ESAT6, a linker, and HspX fused either to mouse Fcγ2a (ESAT6:HspX:mFcγ2a) or 6× His-tag (ESAT6:HspX:His) were synthesized. The resulting proteins were then produced in Pichia pastoris. The fusion proteins were separately emulsified in dimethyldioctadecylammonium bromide(DDA)-trehalose-6,6-dibehenate(TDB) adjuvant, and their immunogenicity with and without bacille Calmette-Guérin (BCG) was assessed in C57BL/6 mice. Th1, Th2, Th17, and T-reg cytokine patterns were evaluated using the ELISA method. Both multistage vaccines induced very strong IL-12 and IFN-γ secretion from splenic cells; the Fc-tagged subunit vaccine induced a more effective Th1 immune response (IFN-γ, 910pg/mL, and IL-12, 854pg/mL) with a very low increase in IL-17 (∼0.1pg/mL) and IL-4 (37pg/mL) and a mild increase in TGF-β (543pg/mL) compared to the BCG or ESAT6:HspX:His primed and boosted groups. The production of IFN-γ to ESAT6:HspX:Fcγ2a was very consistent and showed an increasing trend for IL-12 compared to the BCG or ESAT6:HspX:His primed and boosted groups. Fcγ2a used as a delivery vehicle supported the idea of selective uptake, inducing cross-presentation and forming a proper anti-tuberculosis response in context of Th1/Th2 and Th17/T-reg balances, which is important for protection and prevention of damage. Springer-Verlag Berlin Heidelberg, 2015 Tuberculosis nationallicence APC-targeting nationallicence Fc-fusion vaccine nationallicence ESAT-6 nationallicence HspX nationallicence Soleimanpour Saman Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut Farsiani Hadi Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran aut Mosavat Arman Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut Ghazvini Kiarash Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut Eydgahi Mohammad Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran aut Sankian Mojtaba Immunobiochemistry Lab, Immunology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut Sadeghian Hamid Organic Chemistry, Department of Laboratory Sciences, Mashhad University of Medical Sciences, Mashhad, Iran aut Meshkat Zahra Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut Rezaee Seyed Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/24(2015-12-01), 10467-10480 0175-7598 99:24<10467 2015 99 253 https://doi.org/10.1007/s00253-015-6952-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6952-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Soleimanpour Saman Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 700 1- Farsiani Hadi Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran aut NATIONALLICENCE 700 1- Mosavat Arman Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 700 1- Ghazvini Kiarash Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 700 1- Eydgahi Mohammad Biotechnology Research Center, Semnan University of Medical Sciences, Semnan, Iran aut NATIONALLICENCE 700 1- Sankian Mojtaba Immunobiochemistry Lab, Immunology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 700 1- Sadeghian Hamid Organic Chemistry, Department of Laboratory Sciences, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 700 1- Meshkat Zahra Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 700 1- Rezaee Seyed Microbiology & Virology Research Center, Bu-Ali Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/24(2015-12-01), 10467-10480 0175-7598 99:24<10467 2015 99 253