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Published on: June 1, 2022
Can Anopheles gambiae be infected with Wolbachia pipientis? Insights from an in vitro system
Jason L Rasgon1, Xiaoxia Ren, Michael Petridis
1The 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. jrasgon@jhsph.edu
Abstract:
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.
Insights
Wolbachia pipientis bacteria can infect Anopheles gambiae mosquito cells in vitro. This finding suggests that introducing Wolbachia into Anopheles mosquitoes for malaria control may be possible.
Area of Science:
- Medical entomology
- Microbiology
- Genetics
Background:
- Wolbachia pipientis are endosymbionts that can drive genetic traits in mosquitoes.
- Cytoplasmic incompatibility mediated by Wolbachia is a strategy for transgenic trait introgression in mosquito populations, including for malaria control.
- Anopheles mosquitoes, vectors of malaria, have not been observed to harbor Wolbachia infections.
Purpose of the Study:
- To investigate the susceptibility of Anopheles gambiae cells to Wolbachia pipientis infection using an in vitro system.
- To determine if Anopheles gambiae cells can maintain Wolbachia pipientis infections.
Main Methods:
- An immunocompetent Anopheles gambiae cell line (Sua5B) was used for in vitro infection experiments.
- Wolbachia pipientis strains wRi and wAlbB were introduced into the cell line.
- Infection confirmation involved PCR, antibiotic curing, DNA sequencing, and fluorescence in situ hybridization (FISH).
Main Results:
- Successful establishment and maintenance of Wolbachia pipientis infections (strains wRi and wAlbB) in Anopheles gambiae Sua5B cells.
- Infections were sustained through over 30 passages, indicating stability.
- Confirmation of infection through multiple molecular and microscopic methods.
Conclusions:
- Anopheles gambiae cells are capable of harboring Wolbachia pipientis infections in vitro.
- There is no intrinsic genetic barrier preventing Wolbachia infection in Anopheles gambiae at the cellular level.
- These findings support the feasibility of establishing in vivo Wolbachia infections in Anopheles mosquitoes for potential malaria control strategies.
