Rasgon Jason L, Ren Xiaoxia, Petridis Michael
The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, E4626, 615 N. Wolfe Street, Baltimore, MD 21205, USA.
Appl Environ Microbiol. 2006 Dec;72(12):7718-22. doi: 10.1128/AEM.01578-06. Epub 2006 Oct 6.
Wolbachia pipientis are maternally inherited endosymbionts associated with cytoplasmic incompatibility, a potential mechanism to drive transgenic traits into Anopheles populations for malaria control. W. pipientis infections are common in many mosquito genera but have never been observed in any Anopheles species, leading to the hypothesis that Anopheles mosquitoes are incapable of harboring infection. We used an in vitro system to evaluate the ability of Anopheles gambiae cells to harbor diverse W. pipientis infections. We successfully established W. pipientis infections (strains wRi and wAlbB) in the immunocompetent Anopheles gambiae cell line Sua5B. Infection was confirmed by PCR, antibiotic curing, DNA sequencing, and direct observation using fluorescence in situ hybridization. The infections were maintained at high passage rates for >30 passages. Our results indicate that there is no intrinsic genetic block to W. pipientis infection in A. gambiae cells, suggesting that establishment of in vivo W. pipientis infections in Anopheles mosquitoes may be feasible.
嗜吞噬细胞无形体是一种通过母体遗传的内共生菌，与细胞质不相容性有关，这是一种将转基因性状引入按蚊种群以控制疟疾的潜在机制。嗜吞噬细胞无形体感染在许多蚊子属中很常见，但从未在任何按蚊物种中观察到，这导致了一种假设，即按蚊无法携带感染。我们使用体外系统评估冈比亚按蚊细胞携带不同嗜吞噬细胞无形体感染的能力。我们在具有免疫能力的冈比亚按蚊细胞系Sua5B中成功建立了嗜吞噬细胞无形体感染（菌株wRi和wAlbB）。通过PCR、抗生素消除、DNA测序以及使用荧光原位杂交的直接观察来确认感染。这些感染以高传代率维持了超过30代。我们的结果表明，冈比亚按蚊细胞中不存在对嗜吞噬细胞无形体感染的内在遗传障碍，这表明在按蚊体内建立嗜吞噬细胞无形体感染可能是可行的。
