Dataset Information

New Cell Lines Derived from Laboratory Colony Triatoma infestans and Rhodnius prolixus, Vectors of Trypanosoma cruzi, Do Not Harbour Triatoma Virus.

ABSTRACT: Triatomine bugs of the genera Triatoma and Rhodnius are vectors of Chagas disease, a neglected tropical disease of humans in South America caused by Trypanosoma cruzi. Triatoma virus (TrV), a natural pathogen of Triatoma infestans, has been proposed as a possible tool for the bio-control of triatomine bugs, but research into this virus has been hampered by a lack of suitable host cells for in vitro propagation. Here we report establishment and partial characterisation of continuous cell lines from embryos of T. infestans (TIE/LULS54) and Rhodnius prolixus (RPE/LULS53 and RPE/LULS57). RNAseq screening by a sequence-independent, single primer amplification approach confirmed the absence of TrV and other RNA viruses known to infect R. prolixus, indicating that these new cell lines could be used for propagation of TrV.

SUBMITTER: Penrice-Randal R

PROVIDER: S-EPMC9603895 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature

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Publications

New Cell Lines Derived from Laboratory Colony Triatoma infestans and Rhodnius prolixus, Vectors of Trypanosoma cruzi, Do Not Harbour Triatoma Virus.

Penrice-Randal Rebekah R Hartley Catherine C Beliavskaia Alexandra A Dong Xiaofeng X Brandner-Garrod Luke L Whitten Miranda M Bell-Sakyi Lesley L

Insects 20221005 10

Triatomine bugs of the genera Triatoma and Rhodnius are vectors of Chagas disease, a neglected tropical disease of humans in South America caused by Trypanosoma cruzi. Triatoma virus (TrV), a natural pathogen of Triatoma infestans, has been proposed as a possible tool for the bio-control of triatomine bugs, but research into this virus has been hampered by a lack of suitable host cells for in vitro propagation. Here we report establishment and partial character ...[more]

PMID: 36292854

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Project description:Reactivation of Trypanosoma cruzi transmission by native vectors with different domiciliation capabilities is a major concern for Chagas disease control programs. T. cruzi transmission via intra-domestic Rhodnius prolixus was certified as interrupted by the Pan American Health Organization in Miraflores municipality (Boyacá, Colombia) in 2019. However, Triatoma venosa, a native vector infected with T. cruzi has been increasingly found inside human dwellings across rural areas. In this study, the aim was to describe the eco-epidemiological aspects of T. cruzi transmission in the rural area of Miraflores. For this, we designed a comprehensive, multi-faceted study in 6 rural villages and performed: (i) A cross-sectional serological and molecular study enrolling 155 people and 58 domestic dogs living within 80 households, (ii) a domestic entomological survey, (iii) a determination of the natural infection and blood meal source in collected triatomine bugs, and (iv) an evaluation of synanthropic mammal infection by parasitological and molecular tools. The T. cruzi seroprevalence rates in humans and dogs were 9.03% (14/155) and 22.4% (13/58), respectively. Most infected humans were adults between the ages of 55 and 85 years old. No evidence of T. cruzi DNA was found using qPCR in human blood samples, but we found high parasitemia levels in the infected dogs. In total, 38 triatomine bugs were collected inside dwellings and peridomestic areas: 68.4% (26/38) Triatoma venosa, 29% (11/38) Panstrongylus geniculatus, and 2.6% (1/38) P. rufotuberculatus. Natural infection prevalence was 88% (22/25) for T. venosa, 100% (12/12) for P. geniculatus, and 100% (1/1) P. rufotuberculatus: only TcI was found. No evidence of R. prolixus was found in the area. Two feeding sources were identified in T. venosa (humans and cats), while P. geniculatus fed on cows and bats. Lastly, seven D. marsupialis were captured in peridomestic areas, three were infected with T. cruzi (TcI). The results suggest the existence of T. cruzi transmission cycle between triatomines, dogs, and opossums representing a risk of infection for the human population in rural areas of Miraflores. Despite PAHO declaring Miraflores municipality, Colombia an area of T. cruzi transmission interruption in 2019, this study documents evidence of a secondary vector establishing in domestic settings. T. venosa entomological surveillance is warranted to evaluate prospective human transmission risk in an otherwise 'no-risk' perceived Chagas disease region.

Project description:BackgroundTrypanosoma cruzi is the etiological agent of Chagas' disease. During the parasite life cycle, many molecules are involved in the differentiation process and infectivity. Peptidases are relevant for crucial steps of T. cruzi life cycle; as such, it is conceivable that they may participate in the metacyclogenesis and interaction with the invertebrate host.Methodology/principal findingsIn this paper, we have investigated the effect of the calpain inhibitor MDL28170 on the attachment of T. cruzi epimastigotes to the luminal midgut surface of Rhodnius prolixus, as well as on the metacyclogenesis process and ultrastructure. MDL28170 treatment was capable of significantly reducing the number of bound epimastigotes to the luminal surface midgut of the insect. Once the cross-reactivity of the anti-Dm-calpain was assessed, it was possible to block calpain molecules by the antibody, leading to a significant reduction in the capacity of adhesion to the insect guts by T. cruzi. However, the antibodies were unable to interfere in metacyclogenesis, which was impaired by the calpain inhibitor presenting a significant reduction in the number of metacyclic trypomastigotes. The calpain inhibitor also promoted a direct effect against bloodstream trypomastigotes. Ultrastructural analysis of epimastigotes treated with the calpain inhibitor revealed disorganization in the reservosomes, Golgi and plasma membrane disruption.Conclusions/significanceThe presence of calpain and calpain-like molecules in a wide range of organisms suggests that these proteins could be necessary for basic cellular functions. Herein, we demonstrated the effects of MDL28170 in crucial steps of the T. cruzi life cycle, such as attachment to the insect midgut and metacyclogenesis, as well as in parasite viability and morphology. Together with our previous findings, these results help to shed some light on the functions of T. cruzi calpains. Considering the potential roles of these molecules on the interaction with both invertebrate and vertebrate hosts, it is interesting to improve knowledge on these molecules in T. cruzi.

Dataset Information

New Cell Lines Derived from Laboratory Colony Triatoma infestans and Rhodnius prolixus, Vectors of Trypanosoma cruzi, Do Not Harbour Triatoma Virus.

Publications

New Cell Lines Derived from Laboratory Colony <i>Triatoma infestans</i> and <i>Rhodnius prolixus</i>, Vectors of <i>Trypanosoma cruzi</i>, Do Not Harbour Triatoma Virus.

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