Antiserum against perimicrovillar membranes and midgut tissue reduces the development of Trypanosoma cruzi in the insect vector, Rhodnius prolixus

Gonzalez, Marcelo S.; Hamedi, Afsaneh; Albuquerque-Cunha, José M.; Nogueira, Nadir F. S.; Souza, Wanderley de; Ratcliffe, Norman A.; Azambuja, Patrı́cia; Garcia, Elói S.; Mello, C.B.

doi:10.1016/j.exppara.2006.04.009

Cited by 29 publications

(16 citation statements)

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“…Although we have previously described that HBPs from flagella have more affinity to bind to heparin [27], the data presented here prove that HBPs from both flagella and membrane fractions are able to participate in the adhesion of promatigotes to the Lulo cell monolayers, interacting with heparan sulfate molecules of these cells. These findings are supported by previous studies that indicate the participation of sulfated glycosaminoglycans in host-parasite recognition processes, as: invasion of hepatocytes by Plasmodium sporozoites through the major surface proteins CSP and TRAP [39,40]; adhesion of Plasmodium sp.-infected erythrocytes to the placenta [41]; invasion of mosquito midgut by Plasmodium falciparum [42]; and, adhesion of Trypanosoma epimastigotes to the gut epithelial cells of Rhodnius prolixus [43,44]. …”

Section: Discussionsupporting

confidence: 84%

Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

et al. 2012

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BackgroundLeishmania (V.) braziliensis is a causative agent of cutaneous leishmaniasis in Brazil. During the parasite life cycle, the promastigotes adhere to the gut of sandflies, to avoid being eliminated with the dejection. The Lulo cell line, derived from Lutzomyia longipalpis (Diptera: Psychodidae), is a suitable in vitro study model to understand the features of parasite adhesion. Here, we analyze the role of glycosaminoglycans (GAGs) from Lulo cells and proteins from the parasites in this event.MethodsFlagellar (Ff) and membrane (Mf) fractions from promastigotes were obtained by differential centrifugation and the purity of fractions confirmed by western blot assays, using specific antibodies for cellular compartments. Heparin-binding proteins (HBP) were isolated from both fractions using a HiTrap-Heparin column. In addition, binding of promastigotes to Lulo cells or to a heparin-coated surface was assessed by inhibition assays or surface plasmon resonance (SPR) analysis.ResultsThe success of promastigotes subcellular fractionation led to the obtainment of Ff and Mf proteins, both of which presented two main protein bands (65.0 and 55.0kDa) with affinity to heparin. The contribution of HBPs in the adherence of promastigotes to Lulo cells was assessed through competition assays, using HS or the purified HBPs fractions. All tested samples presented a measurable inhibition rate when compared to control adhesion rate (17 ± 2.0% of culture cells with adhered parasites): 30% (for HS 20μg/ml) and 16% (for HS 10μg/ml); HBP Mf (35.2% for 10μg/ml and 25.4% for 20μg/ml) and HBP Ff (10.0% for 10μg/ml and 31.4% for 20μg/ml). Additionally, to verify the presence of sulfated GAGs in Lulo cells surface and intracellular compartment, metabolic labeling with radioactive sulfate was performed, indicating the presence of an HS and chondroitin sulfate in both cell sections. The SPR analysis performed further confirmed the presence of GAGs ligands on L. (V.) braziliensis promastigote surfaces.ConclusionsThe data presented here point to evidences that HBPs present on the surface of L. (V.) braziliensis promastigotes participate in adhesion of these parasites to Lulo cells through HS participation.

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Section: Discussionsupporting

confidence: 84%

Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

et al. 2012

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“…Once in the gut of the vector, the parasites multiply as epimastigotes forms and migrate to the hindgut where the metacyclogenesis takes place. This process (epimastigotes change to trypomastigotes forms) is important for vertebrate host infection (Gonzalez et al 2006). Many factors have contributed to the presence of trypanosomes in the gut of the invertebrate host; one of them is the perimicrovillar membrane (PMM), a physical and physiological barrier, which is more evident at 10 days after blood meal and important for T. cruzi development in the insect digestive tract (Nogueira et al 1997;Alves et al 2007).…”

Section: Introductionmentioning

confidence: 99%

“…After this, mainly in the small intestine (posterior part of midgut), the epimastigotes divide by binary division and can attach to the perimicrovillar membrane on the intestinal cells that act as an adhesion site for T. cruzi epimastigotes. This process is one of the several important factors for the establishment of the parasite in the vector insect (Alves et al 2007;Gonzalez et al 2006). After this, epimastigotes can attach to the rectal cuticle in the rectum to transform into metacyclic (Kollien and Schaub 2000).…”

Section: Introductionmentioning

confidence: 99%

In vitro and in vivo documentation of quantum dots labeled Trypanosoma cruzi–Rhodnius prolixus interaction using confocal microscopy

et al. 2009

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Semiconductor quantum dots (QDs) are highly fluorescent nanocrystals markers that allow long photobleaching and do not destroy the parasites. In this paper, we used fluorescent core shell quantum dots to perform studies of live parasite-vector interaction processes without any observable effect on the vitality of parasites. These nanocrystals were synthesized in aqueous medium and physiological pH, which is very important for monitoring live cells activities, and conjugated with molecules such as lectins to label specific carbohydrates involved on the parasite-vector interaction. These QDs were successfully used for the study of in vitro and in vivo interaction of Trypanosoma cruzi and the triatomine Rhodnius prolixus. These QDs allowed us to acquire real time confocal images sequences of live T. cruzi-R. prolixus interactions for an extended period, causing no damage to the cells. By zooming to the region of interest, we have been able to acquire confocal images at the three to four frames per second rate. Our results show that QDs are physiological fluorescent markers capable to label living parasites and insect vector cells. QDs can be functionalized with lectins to specifically mark surface carbohydrates on perimicrovillar membrane of R. prolixus to follow, visualize, and understand interaction between vectors and its parasites in real-time.

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“…R. prolixus is an important triatomine vector of Trypanosoma cruzi , the etiologic agent of Chagas disease in Latin America [22-24]. In the insect vector, T. cruzi remains exclusively inside the R. prolixus gut where, in order to survive, the parasite counteracts various host defence factors, including the AMPs [12].…”

Section: Introductionmentioning

confidence: 99%

Humoral responses in Rhodnius prolixus: bacterial feeding induces differential patterns of antibacterial activity and enhances mRNA levels of antimicrobial peptides in the midgut

et al. 2014

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BackgroundThe triatomine, Rhodnius prolixus, is a major vector of Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. It has a strictly blood-sucking habit in all life stages, ingesting large amounts of blood from vertebrate hosts from which it can acquire pathogenic microorganisms. In this context, the production of antimicrobial peptides (AMPs) in the midgut of the insect is vital to control possible infection, and to maintain the microbiota already present in the digestive tract.MethodsIn the present work, we studied the antimicrobial activity of the Rhodnius prolixus midgut in vitro against the Gram-negative and Gram-positive bacteria Escherichia coli and Staphylococcus aureus, respectively. We also analysed the abundance of mRNAs encoding for defensins, prolixicin and lysozymes in the midgut of insects orally infected by these bacteria at 1 and 7 days after feeding.ResultsOur results showed that the anterior midgut contents contain a higher inducible antibacterial activity than those of the posterior midgut. We observed that the main AMP encoding mRNAs in the anterior midgut, 7 days after a blood meal, were for lysozyme A, B, defensin C and prolixicin while in the posterior midgut lysozyme B and prolixicin transcripts predominated.ConclusionOur findings suggest that R. prolixus modulates AMP gene expression upon ingestion of bacteria with patterns that are distinct and dependent upon the species of bacteria responsible for infection.

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Antiserum against perimicrovillar membranes and midgut tissue reduces the development of Trypanosoma cruzi in the insect vector, Rhodnius prolixus

Cited by 29 publications

References 50 publications

Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

Participation of heparin binding proteins from the surface of Leishmania (Viannia) braziliensis promastigotes in the adhesion of parasites to Lutzomyia longipalpis cells (Lulo) in vitro

In vitro and in vivo documentation of quantum dots labeled Trypanosoma cruzi–Rhodnius prolixus interaction using confocal microscopy

Humoral responses in Rhodnius prolixus: bacterial feeding induces differential patterns of antibacterial activity and enhances mRNA levels of antimicrobial peptides in the midgut

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