Recombinant Lactobacillus casei expressing Clostridium perfringens toxoids α, β2, ε and β1 gives protection against Clostridium perfringens in rabbits

被引：14
|
作者：
Zhao, Li [1 ]
Guo, Zhihou [1 ]
Liu, Jiali [1 ]
Wang, Zi [1 ]
Wang, Ruichong [2 ]
Li, Yijing [1 ]
Wang, Li [1 ]
Xu, Yigang [1 ]
Tang, Lijie [1 ]
Qiao, Xinyuan [1 ]
机构：
[1] Northeast Agr Univ, Heilongjiang Key Lab Anim Dis Control & Pharmaceu, Dept Prevent Vet, Coll Vet, Harbin, Heilongjiang, Peoples R China
[2] Heilongjiang Prov Ctr Dis Control & Prevent, Dept Radiol & Protect, Harbin, Heilongjiang, Peoples R China
来源：
VACCINE | 2017年 / 35卷 / 32期
基金：
中国国家自然科学基金;
关键词：
Clostridium perfringens; Alpha-Beta2-Epsilon-Beta1Toxin; Lactobacillus casei; Mucosal immunization; LACTIC-ACID BACTERIA; SITE-DIRECTED MUTAGENESIS; ORAL IMMUNIZATION; MUCOSAL DELIVERY; IMMUNE-RESPONSE; TOXIN; VACCINE; MICE; MUTANT; ANTIGEN;
D O I：
10.1016/j.vaccine.2017.05.076
中图分类号：
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号：
100102 ;
摘要：
The present study used Lactobacillus casei ATCC 393 as antigen delivery system to express C perfringens toxoids alpha-beta 2-epsilon-beta 1 to construct the recombination Lactobacillus casei pPG-2-alpha-beta 2-epsilon-beta 1/L. casei 393. After being induced by 1% xylose, the specificity and integrity of recombinant strain were determined by Western-blotting. Rabbits as native animal model were immunized orally with pPG-2-alpha-beta 2-epsilon-beta 1/L. casei 393 and the titers of specific IgG and sIgA were determined by ELISA. The result showed that oral administration with the recombinants could elicit both local mucosal and systemic immune responses. The proliferation of spleen lymphocytes in rabbits immunized with pPG-2-alpha-beta 2-epsilon-beta/L. casei 393 was observed. Levels of IL-4 and IFN-gamma produced were significantly higher in lymphocytes isolated from the vaccine group than those from the control groups. Flow cytometry assay showed that both the percentages of CD4 + T cells and CD8 + T cells from the vaccine group were significantly increased than the control groups. All these results showed that immunizing with recombinants can elicit both humoral immunity and cellular immunity. Besides, in order to determine the effectiveness of oral immunization with pPG-2-alpha-beta 2-epsilon-beta 1/L. casei 393, rabbits of vaccine group and control groups were challenged with 1 x LD100 unit of culture filtrate of C. perfringens type C and type D toxins respectively. After challenge, 100% of the immunized rabbits survived, while the rabbits of the control group were killed within 48 h. Observation on histopathology showed that histopathological changes were obviously found in heart, liver, spleen, lung, kidney, intestine and brain of rabbits from the control groups, while no apparent histopathological change was observed in the vaccine group. All the results show that pPG-2-alpha-beta 2-epsilon-beta 1/L. casei 393 can elicit effective immunoprotection against C. perfringens. All of these suggest that the use of pPG-2-alpha-beta 2-epsilon-beta 1/L. casei 393 can be regarded as candidate for the development of a vaccine against C. perfringens. (C) 2017 Elsevier Ltd. All rights reserved.
引用
收藏
页码：4010 / 4021
页数：12
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