Establishment of Recombinant Eimeria acervulina Expressing Multi-Copies M2e Derived from Avian Influenza Virus H9N2.
Establishment of Recombinant Eimeria acervulina Expressing Multi-Copies M2e Derived from Avian Influenza Virus H9N2.
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表达禽流感病毒 H9N2 多拷贝 M2e 的重组弯艾美耳球虫的建立
DOI：
10.3390/vaccines9070791
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发表时间：
2021-07-16
期刊：
影响因子：
7.8
通讯作者：
Liu X
中科院分区：
文献类型：
--
作者：
Zhang S;Tang X;Wang S;Shi F;Duan C;Bi F;Suo J;Hu D;Liu J;Wang C;Suo X;Liu X
艾美耳球虫寄生虫作为活疫苗载体的潜力已经报道，在几个物种上成功地进行了遗传操作，如E。tenella、E.美球虫和死灵在感染鸡的7种艾美耳球虫中，E.堆形菌属是一种高度流行的、中等致病性的物种。因此，它是研究转染和作为潜在的疫苗载体使用的价值。在本研究中，使用含有增强型黄色荧光蛋白（EYFP）、红色荧光蛋白（RFP）和12个拷贝的H9N2禽流感病毒M2（M2e）蛋白胞外结构域的表达盒的质粒进行转染。将核转染的子孢子通过翅静脉接种到鸡体内。重组大肠从接种的鸟的粪便中收集具有0.1%EYFP+和RFP+群体的堆型卵囊。经乙胺嘧啶体内筛选和荧光激活细胞分选（FACS）检测，转基因疟原虫的荧光率达到95%以上。Western blot证实M2e在转基因寄生虫（EaM2e）中表达，间接免疫荧光法（IFA）显示M2e在子孢子中的胞质定位。同时，我们发现EaM2e的繁殖力与野生型E相当。堆型（EaWT）。因此，E.成功地建立了堆型。今后的研究重点将放在转基因大肠杆菌是否具有生物学活性上。堆形菌可以作为活疫苗载体。
The potential of Eimeria parasites as live vaccine vectors has been reported with successful genetic manipulation on several species like E. tenella, E. mitis and E. necatrix. Among seven Eimeria species infecting chickens, E. acervulina is a highly prevalent, moderately pathogenic species. Thus, it is valuable for the study of transfection and for use as a potential as vaccine vector. In this study, a plasmid containing expression cassette with enhanced yellow fluorescent protein (EYFP), red fluorescent protein (RFP) and 12 copies of extracellular domain of H9N2 avian influenza virus M2 (M2e) protein was used for the transfection. Nucleofected sporozoites were inoculated into birds through wing vein. Recombinant E. acervulina oocysts with 0.1% EYFP+ and RFP+ populations were collected from the feces of the inoculated birds. The fluorescent rate of transgenic parasites reached over 95% after nine successive propagations with a pyrimethamine selection in vivo and fluorescent-activated cell sorting (FACS) of progeny oocysts. The expression of M2e in the transgenic parasites (EaM2e) was confirmed by Western blot and its cytoplasm localization in sporozoites was displayed by an indirect immunofluorescent assay (IFA). Meanwhile, we found that the fecundity of EaM2e was equivalent to that of wild type E. acervulina (EaWT). Taken together, the stable transfection of E. acervulina was successfully established. Future studies will focus on whether transgenic E. acervulina can serve as a live vaccine vector.
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影响因子：
1.5
作者：
Kelleher, M;Tomley, FM
通讯作者：
Tomley, FM
DOI：
10.1016/j.ijpddr.2014.10.002
发表时间：
2014-12
影响因子：
4
作者：
Chapman, H. David;Jeffers, Thomas K.
通讯作者：
Jeffers, Thomas K.
影响因子：
3.7
作者：
Kim JH;Lee SR;Li LH;Park HJ;Park JH;Lee KY;Kim MK;Shin BA;Choi SY
通讯作者：
Choi SY
表达最大艾美耳球虫抗原的尖复门艾美耳球虫的新型疫苗递送模型可保护鸡免受两种寄生虫的感染
影响因子：
7.3
作者：
Tang X;Liu X;Yin G;Suo J;Tao G;Zhang S;Suo X
通讯作者：
Suo X
影响因子：
7.3
作者：
NORTON, AJ;JORDAN, S;YEOMANS, P
通讯作者：
YEOMANS, P