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Interleukin-4 (IL-4), an immunomodulatory cytokine derived from activated T lymphocytes were shown to regulate Th2-type immune responses. It plays an important role in anti-parasitic infections. In this study, a recombinant plasmid was designed using murine IL-4 co-expressed with pgsA anchor system of Lactobacillus plantarum NC8 and was delivered by live Lactobacillus plantarum NC8, which exhibited an enhanced immunogenicity in protection of BALB/c mice from infection with Trichinella spiralis. The results showed that the levels of serum IgG1 and mucosal secretory IgA (sIgA) were both increased significantly in mice orally inoculated with NC8-pSIP409-pgsA-mIL-4, and the Th2 phenotype immune response was up-regulated. A 29.9 % reduction in adult worm burden at 7 days post-infection (dpi) and 83.3 % reduction in muscle larvae burden at 28 dpi were observed in immune-stimulated mice with NC8-pSIP409-pgsA-mIL-4. Moreover, weight loss and pathological changes were also improved in mice of NC8-pSIP409-pgsA-mIL-4 group. Taken together, it suggests that NC8-pSIP409-pgsA-mIL-4 could improve the intestinal mucosal immunity and promoted the elimination of the adult worm in Trichinella-infected mice. This study laid the foundation for the development of a novel vaccines against Trichinellosis.

Trichinellosis is a very important food-borne parasitic disease, that seriously endangers animal husbandry and food safety. Therefore, it is necessary to develop a safe and effective vaccine against Trichinella spiralis infection. In this experiment, invasive Lactobacillus plantarum carrying the FnBPA gene served as a live bacterial vector to deliver nucleic acids to the host to produce a novel oral nucleic acid vaccine. Coexpression of the T. spiralis cathepsin F-like protease 1 gene (TsCPF1) and murine IL-4 (mIL-4) by the nucleic acid vaccine was constructed and subsequently delivered to intestinal epithelial cells via invasive L. plantarum. Thirty-seven days after the first immunization, the experimental mice were challenged with 350 T. spiralis infective larvae by oral gavage. The results showed that mice orally immune-stimulated with invasive L. plantarum pValac-TsCPF1/pSIP409-FnBPA not only produce anti-TsCPF1-specific IgG antibodies, sIgA, Th1/Th2 cytokine distinctly increased but also intestinal damage and worm burden relieved compare to non-invasive TsCPF1 group (pValac-TsCPF1/pSIP409). Most notably, experimental mice immunized with invasive L. plantarum coexpressing TsCPF1 and mIL-4 (pValac-TsCPF1-IL-4/pSIP409-FnBPA) exhibited the highest protection efficiency against T. spiralis infection. The above results reveal that invasive L. plantarum-expressing the FnBPA protein improved mucosal and cellular immunity and enhanced resistance to T. spiralis. The nucleic acid vaccine delivered by invasive L. plantarum described in this study offers a novel idea for the prevention of T. spiralis.

The coccidiosis caused by Eimeria tenella (coccidian) and other species is a serious parasitic disease that affects the global poultry breeding industry. Lactobacillus strains exhibit a number of properties that make them attractive candidates as delivery vehicles for presentation to the mucosa of compounds with pharmaceutical interest, particularly vaccines. Here, the recombinant Lactobacillus plantarum (co-expressing SO7 and DCpep gene) was constructed, and its efficacy against E. tenella challenge was evaluated in this study. Broiler chickens were orally immunized with live recombinant L. plantarum NC8-pSIP409-SO7-DCpep for two weeks and were then challenged with 5×104E.tenella sporulated oocysts per chicken. During the experiment, body weight gains, cecum lesion scores, fecal oocyst shedding and antibody responses in serum and intestinal washes were assessed as measures of protective immunity. The results indicated that chickens immunized with live recombinant L. plantarum can increase body weight gains and serum antibody responses compared to the control groups. Meanwhile, fecal oocyst shedding in the immunized group was significantly reduced (p

Coccidiosis is an intestinal parasitic disease that is caused by Eimeria tenella and other species, and it seriously restricts the economic development of the broiler breeding industry. In this study, a recombinant Lactobacillus plantarum with an invasive effect was constructed, and it expressed the TA4-AMA1 protein of E. tenella. After oral immunization with recombinant L. plantarum, specific humoral and mucosal immune levels were measured by indirect ELISA, and the differentiation of T cells was analysed by flow cytometry. After challenge with sporulated oocysts, the body weight, oocyst shedding and cecum lesions of the chicken were evaluated. The results indicated that chickens immunized with recombinant invasive L. plantarum produced higher levels of specific antibodies in the serum than did the non-immunized controls, and the secretory IgA (sIgA) levels were increased in the intestinal washes compared to those of the controls (P < 0.05). Flow cytometry showed that recombinant invasive L. plantarum significantly stimulated T cell differentiation compared to the PBS group (P < 0.01, P < 0.001), and a higher proportion of CD4+ and CD8+ T cells were detected in peripheral blood. Moreover, the lesion scores and histopathological caecum sections showed that immunizing chickens with recombinant invasive L. plantarum can significantly relieve pathological damage in the cecum (P < 0.01), and the relative body weight gain was 89.64 %, which was higher than the 79.83 % gain in the chickens immunized with non-invasive L. plantarum. After the challenge, faeces from ten chickens in each group were collected between 4 and 7 days, and the oocysts per gram (OPG) was determined by the McMaster technique. The data indicated that oocysts in the faeces of chickens immunized with the recombinant invasive L. plantarum were significantly lower than those of the controls (P < 0.01). The results suggest that recombinant invasive L. plantarum effectively activated immune responses against E. tenella infection and can be used as a candidate vaccine against E. tenella infection.