Chimeric symbionts expressing a Wolbachia protein stimulate mosquito immunity and inhibit filarial parasite development

Epis, Sara; Varotto-Boccazzi, Ilaria; Crotti, Elena; Damiani, Claudia; Giovati, Laura; Mandrioli, Mauro; Biggiogera, Marco; Gabrieli, Paolo; Genchi, Marco; Polonelli, Luciano; Daffonchio, Daniele; Favia, Guido; Bandi, Claudio

doi:10.1038/s42003-020-0835-2

Cited by 27 publications

(28 citation statements)

References 63 publications

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“…native host, culturable microorganisms can be directly exploited in different biotechnological applications (Crotti et al, 2012;Valiente Moro et al, 2013;Epis et al, 2020). For instance, they have been proposed as probiotics for honeybees to sustain insect health against specific pathogens (Crotti et al, 2013;Daisley et al, 2019) or as beneficial strains to improve the fitness of sterile male insects vs. conventional ones (Ben Ami et al, 2010;Augustinos et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Hydrolytic Profile of the Culturable Gut Bacterial Community Associated With Hermetia illucens

et al. 2020

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Larvae of the black soldier fly (BSF) Hermetia illucens (L.) convert organic waste into high valuable insect biomass that can be used as alternative protein source for animal nutrition or as feedstock for biodiesel production. Since insect biology and physiology are influenced by the gut microbiome, knowledge about the functional role of BSFassociated microorganisms could be exploited to enhance the insect performance and growth. Although an increasing number of culture-independent studies are unveiling the microbiota structure and composition of the BSF gut microbiota, a knowledge gap remains on the experimental validation of the contribution of the microorganisms to the insect growth and development. We aimed at assessing if BSF gut-associated bacteria potentially involved in the breakdown of diet components are able to improve host nutrition. A total of 193 bacterial strains were obtained from guts of BSF larvae reared on a nutritious diet using selective and enrichment media. Most of the bacterial isolates are typically found in the insect gut, with major representatives belonging to the Gammaproteobacteria and Bacilli classes. The hydrolytic profile of the bacterial collection was assessed on compounds typically present in the diet. Finally, we tested the hypothesis that the addition to a nutritionally poor diet of the two isolates Bacillus licheniformis HI169 and Stenotrophomonas maltophilia HI121, selected for their complementary metabolic activities, could enhance BSF growth. B. licheniformis HI169 positively influenced the larval final weight and growth rate when compared to the control. Conversely, the addition of S. maltophilia HI121 to the nutritionally poor diet did not result in a growth enhancement in terms of larval weight and pupal weight and length in comparison to the control, whereas the combination of the two strains positively affected the larval final weight and the pupal weight and length. In conclusion, we isolated BSF-associated bacterial strains with potential positive properties for the host nutrition and we showed that selected isolates may enhance BSF growth, suggesting the importance to evaluate the effect of the bacterial administration on the insect performance.

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Section: Discussionmentioning

confidence: 99%

Hydrolytic Profile of the Culturable Gut Bacterial Community Associated With Hermetia illucens

et al. 2020

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“…The bacterium Asaia WSP derived from the engineering of the bacterium Asaia SF2.1 strain, originally isolated from the Anopheles stephensi mosquito (Favia et al, 2007), with the plasmid pHM4-WSP (Epis et al, 2020). Briefly, the WSP cassette, inserted in the plasmid pHM4, is composed by the Wolbachia surface protein gene sequence (from the Wolbachia of the nematode Dirofilaria immitis ), the neomycin phosphotranferase promoter PnptII, the E-TAG epitope and the transcription terminator Trrn.…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

“…Briefly, the WSP cassette, inserted in the plasmid pHM4, is composed by the Wolbachia surface protein gene sequence (from the Wolbachia of the nematode Dirofilaria immitis ), the neomycin phosphotranferase promoter PnptII, the E-TAG epitope and the transcription terminator Trrn. Asaia pHM4 was also obtained from strain Asaia SF2.1, but transformed with the empty plasmid (without the WSP-coding gene) and was used as control bacterium (Epis et al, 2020). Both bacteria were grown overnight in GLY medium broth (glycerol 25 g/l and yeast extract 10 g/l, pH 5) supplemented with kanamycin 100 μg/ml, under constant agitation at 30°C.…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

“…In this study, we suggest that bacteria of the genus Wolbachia represent a promising source of molecules capable of stimulating and modulating innate immunity, with the potential to be exploited in immunotherapy and prophylaxis. In insects, Wolbachia has been shown to be a potent activator of innate immunity, able to determine the upregulation of several immune effectors such as antimicrobial peptides, autophagy-related proteins, and ROS (Zug & Hammerstein, 2015;Epis et al, 2020). Indeed, the successful use of Wolbachia to block the transmission of viruses by mosquitoes has in part been associated with this immune-activating capacity (Moreira et al, 2009;Rancès et al, 2012; LePage & Bordenstein, 2013).…”

Section: Introductionmentioning

confidence: 99%

“…For example, Jacouton and colleagues (2019) have modified a Lactococcus lactis strain for the expression of the cytokine IL-17A, with tumor prevention in a mouse model, after intranasal delivery of the engineered bacterium. In this context, we recently selected an acetic acid bacterium, Asaia sp., as a bacterial vehicle for the expression of the Wolbachia surface protein, generating the chimeric symbiont Asaia WSP (Epis et al, 2020). Our hypothesis is that Asaia WSP should determine innate immune activation with a M1 bias, thus conferring protection against M1-impaired infections.…”

Section: Introductionmentioning

confidence: 99%

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Boosting innate immunity:Asaiabacteria expressing a protein fromWolbachiadetermine macrophage activation and killing ofLeishmania

Varotto-Boccazzi

Epis

Arnoldi

et al. 2020

Preprint

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AbstractLeishmaniases are severe vector-borne diseases affecting humans and animals, caused by Leishmania protozoans. Immune polarization plays a major role in determining the outcome of Leishmania infections: hosts displaying M1-polarized macrophages are protected, while those biased on the M2 side acquire a chronic infection, that could develop into an overt and potentially deadly disease. The identification of the factors involved in M1 polarization is essential for the design of therapeutic and prophylactic interventions, including vaccines. Infection by the filarial nematode Dirofilaria immitis could be one of the factors that interfere with leishmaniasis in dogs. Indeed, filarial nematodes induce a partial skew of the immune response towards M1, likely caused by their bacterial endosymbionts, Wolbachia. Here we have examined the potential of AsaiaWSP, a bacterium engineered for the expression of the Wolbachia surface protein (WSP), as an inductor of M1 macrophage activation and Leishmania killing. Macrophages stimulated with AsaiaWSP displayed a strong leishmanicidal activity, comparable to that determined by the choice-drug amphotericin B. Additionally, AsaiaWSP determined the expression of markers of classical macrophage activation, including M1 cytokines, ROS and NO, and an increase in phagocytosis activity. Asaia not expressing WSP also induced macrophage activation, although at a lower extent compared to AsaiaWSP. In summary, our study, while providing a strong evidence for the immune-stimulating properties of Wolbachia, highlights the translational potential of AsaiaWSP in the areas of the immune-prophylaxis and therapy of leishmaniases, as well as of other diseases that could be subverted by M1 macrophage activation.

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On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

Fricke

Klemm

Bott

et al. 2021

Appl Microbiol Biotechnol

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Acetic acid bacteria (AAB) are valuable biocatalysts for which there is growing interest in understanding their basics including physiology and biochemistry. This is accompanied by growing demands for metabolic engineering of AAB to take advantage of their properties and to improve their biomanufacturing efficiencies. Controlled expression of target genes is key to fundamental and applied microbiological research. In order to get an overview of expression systems and their applications in AAB, we carried out a comprehensive literature search using the Web of Science Core Collection database. The Acetobacteraceae family currently comprises 49 genera. We found overall 6097 publications related to one or more AAB genera since 1973, when the first successful recombinant DNA experiments in Escherichia coli have been published. The use of plasmids in AAB began in 1985 and till today was reported for only nine out of the 49 AAB genera currently described. We found at least five major expression plasmid lineages and a multitude of further expression plasmids, almost all enabling only constitutive target gene expression. Only recently, two regulatable expression systems became available for AAB, an N-acyl homoserine lactone (AHL)-inducible system for Komagataeibacter rhaeticus and an l-arabinose-inducible system for Gluconobacter oxydans. Thus, after 35 years of constitutive target gene expression in AAB, we now have the first regulatable expression systems for AAB in hand and further regulatable expression systems for AAB can be expected. Key points • Literature search revealed developments and usage of expression systems in AAB. • Only recently 2 regulatable plasmid systems became available for only 2 AAB genera. • Further regulatable expression systems for AAB are in sight.

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Chimeric symbionts expressing a Wolbachia protein stimulate mosquito immunity and inhibit filarial parasite development

Cited by 27 publications

References 63 publications

Hydrolytic Profile of the Culturable Gut Bacterial Community Associated With Hermetia illucens

Hydrolytic Profile of the Culturable Gut Bacterial Community Associated With Hermetia illucens

Boosting innate immunity:Asaiabacteria expressing a protein fromWolbachiadetermine macrophage activation and killing ofLeishmania

On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

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