Abstract
BackgroundTrichinellosis is a serious zoonotic disease distributed around the world. It is needed to develop a safe, effective and feasible anti-Trichinella vaccine for prevention and control of trichinellosis. The aim of this study was to construct a recombinant Lactobacillus plantarum encoding Trichinella spiralis inorganic pyrophosphatase (TsPPase) and investigate its immune protective effects against T. spiralis infection.Methodology/Principal findingsThe growth of recombinant L. plantarum was not affected by TsPPase/pSIP409-pgsA′ plasmid, and the recombinant plasmid was inherited stably in bacteria. Western blot and immunofluorescence assay (IFA) indicated that the rTsPPase was expressed on the surface of recombinant L. plantarum. Oral vaccination with rTsPPase induced higher levels of specific serum IgG, IgG1, IgG2a and mucosal secretory IgA (sIgA) in BALB/c mice. ELISA analysis revealed that the levels of IFN-γ and IL-4 released from spleen, mesenteric lymph nodes and Peyer’s patches were evidently increased at 2–4 weeks following vaccination, compared to MRS (De Man, Rogosa, Sharpe) medium control group (P < 0.05). Immunization of mice with rTsPPase exhibited a 67.18, 54.78 and 51.91% reduction of intestinal infective larvae, adult worms and muscle larvae at 24 hours post infection (hpi), 6 days post infection (dpi) and 35 dpi, respectively (P < 0.05), and the larval molting and development was significantly inhibited by 45.45% at 24 hpi, compared to the MRS group.ConclusionsTsPPase plays a crucial role in T. spiralis molting and development, oral vaccination with rTsPPase induced a significant local mucosal sIgA response and systemic Th1/Th2 immune response, and immune protection against T. spiralis infection in BALB/c mice.
Highlights
Trichinellosis is mainly caused by the zoonotic nematode Trichinella spiralis which is a tissuedwelling nematode infected over 150 kinds of mammals over the world [1,2]
Trichinella spiralis inorganic pyrophosphatase (TsPPase) plays a crucial role in T. spiralis molting and development, oral vaccination with rTsPPase induced a significant local mucosal secretory IgA (sIgA) response and systemic Th1/Th2 immune response, and immune protection against T. spiralis infection in BALB/c mice
The results of western blot revealed that an individual protein band of recombinant NC8 with about 44.0 kDa was recognized by anti-rTsPPase serum and infection serum, but no bands were recognized in the soluble protein of L. plantarum NC8 (Fig 4)
Summary
Trichinellosis is mainly caused by the zoonotic nematode Trichinella spiralis which is a tissuedwelling nematode infected over 150 kinds of mammals over the world [1,2]. Humans acquire trichinellosis by ingesting raw or semi-raw meat containing Trichinella larvae. There is an urgent need to develop a safe, effective and feasible antiTrichinella vaccine for control of Trichinella infection in food animals and for food safety. Trichinellosis is a serious zoonotic disease distributed around the world. It is needed to develop a safe, effective and feasible anti-Trichinella vaccine for prevention and control of trichinellosis. The aim of this study was to construct a recombinant Lactobacillus plantarum encoding Trichinella spiralis inorganic pyrophosphatase (TsPPase) and investigate its immune protective effects against T. spiralis infection
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