Construction of Lactococcus lactis expressing secreted and anchored Eimeria tenella 3-1E protein and comparison of protective immunity against homologous challenge
Construction of Lactococcus lactis expressing secreted and anchored Eimeria tenella 3-1E protein and comparison of protective immunity against homologous challenge
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DOI：
10.1016/j.exppara.2017.05.001
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发表时间：
2017-07-01
影响因子：
2.1
通讯作者：
Ma, Dexing
中科院分区：
文献类型：
--
作者：
Ma, Chunli;Zhang, Lili;Ma, Dexing
构建了两个新的质粒pTX 8048-SP-Delta 3- 1 E和pTX 8048-SP-NA Delta 3- 1 E-CWA。将质粒分别电转化到乳酸乳杆菌NZ 9000中，以产生3 - 1 E蛋白在分泌物中表达的乳酸乳杆菌/pTX 8048-SP-Delta 3- 1 E和3 - 1 E蛋白共价锚定到细菌细胞表面的乳酸乳杆菌/pTX 8048-SP-NA Delta 3- 1 E-CWA。Western blot检测目的蛋白的表达。将表达分泌型3- 1 E蛋白、锚定型3- 1 E蛋白和胞质型3- 1 E蛋白的活乳球菌分别经口给予鸡，通过动物试验比较其免疫保护力和效力。结果表明，表达锚定3- 1 E蛋白的重组乳球菌经口免疫鸡，可诱导产生较高的3- 1 E特异性IgG，提高脾脏中CD 4+和CD 8 α +细胞的比例，减轻盲肠平均病变评分，降低单鸡卵囊产量。综上所述，这些发现表明，由L lacits展示的表面锚定的艾美耳球虫蛋白可以诱导针对同源感染的保护性免疫和部分保护。(C)2017爱思唯尔公司All rights reserved.
Two novel plasmids pTX8048-SP-Delta 3-1E and pTX8048-SP-NA Delta 3-1E-CWA were constructed. The plasmids were respectively electrotransformed into L lactis NZ9000 to generate strain of L lactis/pTX8048-SP-Delta 3-1E in which 3-1E protein was expressed in secretion, and L lactis/pTX8048-SP-NA Delta 3-1E-CWA on which 3-1E protein was covalently anchored to the surface of bacteria cells. The expression of target proteins were examined by Western blot. The live lactococci expressing secreted 3-1E protein, anchored 3-1E protein, and cytoplasmic 3-1E protein was administered orally to chickens respectively, and the protective immunity and efficacy were compared by animal experiment. The results showed oral immunization to chickens with recombinant lactococci expressing anchored 3-1E protein elicited high 3-1E-specific serum IgG, increased high proportion of CD4(+) and CD8 alpha(+) cells in spleen, alleviated average lesion score in cecum, decreased the oocyst output per chicken compared to lactococci expressing cytoplasmic or secreted 3-1E protein. Taken together, these findings indicated the surface anchored Eimeria protein displayed by L lacits can induce protective immunity and partial protection against homologous infection. (C) 2017 Elsevier Inc. All rights reserved.