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Background: Development of Helicobacter pylori vaccine would be a new effective strategy for prevention and treatment of H. pylori infection. Recombinant H. pylori vaccine comprising a single subunit antigen can only induce immune response with limited protection efficiency. In this study, the protective effect of H. pylori multicomponent vaccines consisting of three recombinant subunit antigens was investigated using the Mongolian gerbil model. Materials and methods: Mongolian gerbils were immunized with different formulations of three recombinant H. pylori antigens (UreB, HspA, and HpaA) with two different adjuvants (Al(OH) 3 , LT R72DITH ) by intragastric (i.g.) or intramuscular (i.m.) routes. The protective effects of multicomponent vaccines were assessed after H. pylori challenge in different studies. The specific IgG antibodies in serum were monitored by ELISA, and the mRNA expressions of IL-4 and IFN- γ in spleen tissue were detected by reverse transcription- polymerase chain reaction (RT-PCR). Results: The protective effect against H. pylori challenge in gerbils immunized with three recombinant antigens and LT R72DITH or Al(OH) 3 was significantly higher than that in single- or double-antigen vaccine-immunized and control gerbils. Furthermore, the protective effect of the triple-antigen vaccine combined with the LT R72DITH adjuvant (average 86.3%) was significantly greater than that of vaccine combined with the Al(OH) 3 adjuvant (average 53.4%). After the first immunization, the anti-UreB/HspA/HpaA serum IgG level in gerbils immunized with triple-antigen vaccine combined with Al(OH) 3 was higher than that in gerbils immunized with the vaccine combined with LT R72DITH . Splenic interferon (IFN)- γ and interleukin (IL)-4 transcript levels were significantly increased in LT R72DITH vaccine-immunized gerbils as compared to the Al(OH) 3 vaccine group. Moreover, splenic IL-4 mRNA levels were higher than IFN- γ in gerbils immunized with triple-antigen vaccine with either LT R72DITH or Al(OH) 3 . Conclusions: This study indicated that the recombinant multicomponent vaccine provided effective protection against H. pylori infection as compared to the single-antigen vaccine. This protective immunity would be closely associated with a predominant Th2-type response.

To observe the effect of moxibustion of "Dachangshu" (BL 25) on pain reaction and expression of transient receptor potential vanilloid 1 (TRPV 1) of bone marrow cells in visceral hyperalgesia (VHA) rats so as to explore its mechanism underlying visceral pain-relief.Twenty-eight male SD rats were divided into control group, control+moxibustion group, VHA model and VHA+moxibustion group (n = 7/group). The VHA model was made by giving colorectal distension (CRD, 60 mmHg) to the newborn rats for 1 min (repeated once again in 30 min) from postnatal day 8 on, once daily for a week. Moxibustion was applied to ipsilateral "Dachangshu"(BL 25) area for 40 min from the 8th week on after birth. Abdominal withdrawal reflex (AWR) and pain threshold during CRD were measured before and after moxibustion. The TRPV 1 mRNA expressio of bone marrow cells was detected by real time-POR.(1) The AWR score of the model group was significantly higher than that of the control group, but the pain threshold of the model group was significantly lower than that of the control group (P0.01), suggesting a VHA in model rats. (2) After moxibustion, the AWR scores were significantly lower in the VHA+moxibustion group than in the model group (P0.05, P0.01), and the pain threshold was remarkably higher in the former group than in the latter group (P0.01). Similar results were found in the control+moxibustion group compared to the control group: the decreased AWR scores (CRD 40 mmHg, 60 mmHg and 80 mmHg, P0.01) and the increased pain threshold (P0.05). (3) The TRPV 1 mRNA expression level of bone marrow cells was significantly lower in the VHA + moxibustion group than in the model group (P0.01). No significant difference was found between the control and moxibustion+control groups in TRPV 1 mRNA expression level (P0.05).Moxibustion of "Dachangshu" (BL 25) can reduce visceral hyperalgesia and down-regulate TRPV 1 mRNA expression of bone marrow cells in VHA rats, suggesting an involvement of TRPV 1 mRNA of bone marrow cells in CRD-induced visceral pain development.

Development of Helicobacter pylori vaccine would be a new effective strategy for prevention and treatment of H. pylori infection. Recombinant H. pylori vaccine comprising a single subunit antigen can only induce immune response with limited protection efficiency. In this study, the protective effect of H. pylori multicomponent vaccines consisting of three recombinant subunit antigens was investigated using the Mongolian gerbil model.Mongolian gerbils were immunized with different formulations of three recombinant H. pylori antigens (UreB, HspA, and HpaA) with two different adjuvants (Al(OH)3, LT(R72DITH)) by intragastric (i.g.) or intramuscular (i.m.) routes. The protective effects of multicomponent vaccines were assessed after H. pylori challenge in different studies. The specific IgG antibodies in serum were monitored by ELISA, and the mRNA expressions of IL-4 and IFN-gamma in spleen tissue were detected by reverse transcribed polymerase chain reaction (RT-PCR).The protective effect against H. pylori challenge in gerbils immunized with three recombinant antigens and LT(R72DITH) or Al(OH)3 was significantly higher than that in single- or double-antigen vaccine-immunized and control gerbils. Furthermore, the protective effect of the triple-antigen vaccine combined with the LT(R72DITH) adjuvant (average 86.3%) was significantly greater than that of vaccine combined with the Al(OH)3 adjuvant (average 53.4%). After the first immunization, the anti-UreB/HspA/HpaA serum IgG level in gerbils immunized with triple-antigen vaccine combined with Al(OH)3 was higher than that in gerbils immunized with the vaccine combined with LT(R72DITH). Splenic interferon (IFN)-gamma and interleukin (IL)-4 transcript levels were significantly increased in LT(R72DITH) vaccine-immunized gerbils as compared to the Al(OH)3 vaccine group. Moreover, splenic IL-4 mRNA levels were higher than IFN-gamma in gerbils immunized with triple-antigen vaccine with either LT(R72DITH) or Al(OH)3.This study indicated that the recombinant multicomponent vaccine provided effective protection against H. pylori infection as compared to the single-antigen vaccine. This protective immunity would be closely associated with a predominant Th2-type response.