Abstract
Recently, we have developed a novel Mycobacterium-Escherichia coli shuttle vector system using pMyong2, which can provide an enhanced expression of heterologous genes in recombinant Mycobacterium smegmatis (rSmeg). To investigate the usefulness of rSmeg using pMyong2 in vaccine application, we vaccinated M. smegmatis with pMyong2 system expressing Human Immunodeficiency Virus Type I (HIV-1) Gag p24 antigen (rSmeg-pMyong2-p24) into mice and examined its cellular and humoral immune responses against HIV gag protein. We found that rSmeg-pMyong2-p24 expressed higher levels of Gag protein in bacteria, macrophage cell line (J774A.1) and mouse bone marrow derived dendritic cells (BMDCs) compared to rSmeg strains using two other vector systems, pAL5000 derived vector (rSmeg-pAL-p24) and the integrative plasmid, pMV306 (rSmeg-pMV306-p24). Inoculation of mice with rSmeg-pMyong2-p24 elicited more effective immunity compared to the other two rSmeg strains, as evidenced by higher levels of HIV-1 Gag-specific CD4 and CD8 T lymphocyte proliferation, interferon gamma ELISPOT cell induction, and antibody production. Furthermore, rSmeg-pMyong2-p24 showed a higher level of cytotoxic T cell response against target cells expressing Gag p24 proteins. Our data suggest that Mycobacterium-Escherichia coli shuttle vector system with pMyong2 may provide an advantage in vaccine application of rSmeg over other vector systems.
Highlights
An effective human immunodeficiency virus (HIV) vaccine will likely need to elicit virus-specific neutralizing antibodies and cytotoxic T-lymphocyte (CTL) responses
To explore the usefulness of pMyong[2] vector system in the generation of recombinant M. smegmatis (rSmeg) strains for HIV type 1 (HIV-1) p24 Gag vaccination, we generated a total of 3 types of rSmeg strains expressing p24, rSmeg-pMyong2-p24, rSmeg-pAL-p24 and rSmeg-pMV306-p24 using different types of Mycobacterium-E. coli shuttle vectors, pMyong2-TOPO28, pAL-TOPO28, and pMV30629, respectively (Fig. 1)
For vaccine development against diseases such as AIDS and tuberculosis, attention has focused on developing strategies for the vaccine induction of cellular immunity, CTL1
Summary
An effective human immunodeficiency virus (HIV) vaccine will likely need to elicit virus-specific neutralizing antibodies and cytotoxic T-lymphocyte (CTL) responses. Mycobacterium bovis BCG (BCG), currently the most widely administered vaccine in the world, is a live attenuated vaccine used to protect against tuberculosis and leprosy[2,3,4,5,6] It demonstrates excellent adjuvant properties, induces long lasting immunity and has a low production cost[7,8,9]. The aim of the present study is to investigate the usefulness of rSmeg with pMyong[2] in HIV vaccine application To this end, we constructed the rSmeg with pMyong[2] system expressing HIV-1 p24 Gag antigen (rSmeg-pMyong2-p24) and examined its cellular and humoral immune responses against HIV Gag proteins in vaccinated mice compared with rSmeg strains transfected with 2 other vector systems, an episomal plasmid, pAL5000 derived vector (rSmeg-pAL-p24) and an integrative plasmid, pMV306 (rSmeg-pMV306-p24)
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