Evaluation of a recombinant protein(rTc24) and synthetic peptides in anti-Trypanosoma cruzi positive samples from blood bank donors in chagasic endemic areas of ecuador.

At least eighteen species of triatominae have been found in the Brazilian Amazon, nine of them naturally infected with Trypanosoma cruzi or "cruzi-like" trypanosomes and associated with numerous wild reservoirs. Despite the small number of human cases of Chagas' disease described to date in the Brazilian Amazon the risk that the disease will become endemic in this area is increasing for the following reasons: a) uncontrolled deforestation and colonization altering the ecological balance between reservoir hosts and wild vectors; b) the adaptation of reservoir hosts of T.cruzi and wild vectors to peripheral and intradomiciliary areas, as the sole feeding alternative; c) migration of infected human population from endemic areas, accompanied by domestic reservoir hosts (dogs and cats) or accidentally carrying in their baggage vectors already adapted to the domestic habitat. In short, risks that Chagas' disease will become endemic to the Amazon appear to be linked to the transposition of the wild cycle to the domestic cycle in that area or to transfer of the domestic cycle from endemic areas to the Amazon.

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Molecular diagnosis of Trypanosoma cruzi/Leishmania spp. coinfection in domestic, peridomestic and wild mammals of Venezuelan co-endemic areas

Veterinary Parasitology Regional Studies and Reports ◽

10.1016/j.vprsr.2018.10.002 ◽

2018 ◽

Vol 14 ◽

pp. 123-130 ◽

Cited By ~ 1

Author(s):

Mercedes Viettri ◽

Leidi Herrera ◽

Cruz M. Aguilar ◽

Antonio Morocoima ◽

Jesús Reyes ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Molecular Diagnosis ◽

Wild Mammals ◽

Leishmania Spp ◽

Endemic Areas

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A prime-boost immunization with Tc52 N-terminal domain DNA and the recombinant protein expressed in Pichia pastoris protects against Trypanosoma cruzi infection

Vaccine ◽

10.1016/j.vaccine.2016.05.011 ◽

2016 ◽

Vol 34 (28) ◽

pp. 3243-3251 ◽

Cited By ~ 11

Author(s):

Marina N. Matos ◽

Andrés Sánchez Alberti ◽

Celina Morales ◽

Silvia I. Cazorla ◽

Emilio L. Malchiodi

Keyword(s):

Pichia Pastoris ◽

Recombinant Protein ◽

Trypanosoma Cruzi ◽

Terminal Domain ◽

Boost Immunization ◽

Cruzi Infection

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Evaluation of Serological Tests To Identify Trypanosoma cruzi Infection in Humans and Determine Cross-Reactivity with Trypanosoma rangeli and Leishmania spp.

Clinical and Vaccine Immunology ◽

10.1128/cvi.00127-07 ◽

2007 ◽

Vol 14 (8) ◽

pp. 1045-1049 ◽

Cited By ~ 99

Author(s):

Zuleima C. Caballero ◽

Octavio E. Sousa ◽

Waldelania P. Marques ◽

Amadeo Saez-Alquezar ◽

Eufrosina S. Umezawa

Keyword(s):

Trypanosoma Cruzi ◽

Synthetic Peptides ◽

Cross Reactivity ◽

Reference Test ◽

Trypanosoma Rangeli ◽

Serological Tests ◽

Western Blot Assay ◽

Leishmania Spp ◽

Cruzi Infection ◽

Secreted Antigens

ABSTRACT Five commercially available enzyme-linked immunosorbent assays (ELISAs), one in-house ELISA, and two hemagglutination assays were evaluated to determine their diagnostic accuracy for Chagas' disease in two studies. In study 1, ELISA kits showed 100% sensitivity, but specificities ranged from 82.84% to 100% when leishmaniasis cases were included and from 95.57% to 100% when leishmaniasis cases were excluded. Kits using recombinant antigens or synthetic peptides are more specific than those using crude extracts from Trypanosoma cruzi epimastigote forms. Kits evaluated in Panama, in study 2, showed 75% to 100% sensitivity and 97.12% to 100% specificity. These data were obtained by using a Western blot assay with T. cruzi trypomastigote excreted-secreted antigens as a reference test to confirm T. cruzi infection.

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CD8+-T-Cell-Dependent Control of Trypanosoma cruzi Infection in a Highly Susceptible Mouse Strain after Immunization with Recombinant Proteins Based on Amastigote Surface Protein 2

Infection and Immunity ◽

10.1128/iai.73.9.6017-6025.2005 ◽

2005 ◽

Vol 73 (9) ◽

pp. 6017-6025 ◽

Cited By ~ 44

Author(s):

Adriano F. S. Araújo ◽

Bruna C. G. de Alencar ◽

José Ronnie C. Vasconcelos ◽

Meire I. Hiyane ◽

Cláudio R. F. Marinho ◽

...

Keyword(s):

Amino Acids ◽

T Cells ◽

T Cell ◽

Recombinant Protein ◽

Trypanosoma Cruzi ◽

Recombinant Proteins ◽

Protective Immunity ◽

Protective Efficacy ◽

Surface Protein ◽

Cpg Oligodeoxynucleotide

ABSTRACT We previously described that DNA vaccination with the gene encoding amastigote surface protein 2 (ASP-2) protects approximately 65% of highly susceptible A/Sn mice against the lethal Trypanosoma cruzi infection. Here, we explored the possibility that bacterial recombinant proteins of ASP-2 could be used to improve the efficacy of vaccinations. Initially, we compared the protective efficacy of vaccination regimens using either a plasmid DNA, a recombinant protein, or both sequentially (DNA priming and protein boosting). Survival after the challenge was not statistically different among the three mouse groups and ranged from 53.5 to 75%. The fact that immunization with a recombinant protein alone induced protective immunity revealed the possibility that this strategy could be pursued for vaccination. We investigated this possibility by using six different recombinant proteins representing distinct portions of ASP-2. The vaccination of mice with glutathione S-transferase fusion proteins representing amino acids 261 to 500 or 261 to 380 of ASP-2 in the presence of the adjuvants alum and CpG oligodeoxynucleotide 1826 provided remarkable immunity, consistently protecting 100% of the A/Sn mice. Immunity was completely reversed by the in vivo depletion of CD8+ T cells, but not CD4+ T cells, and was associated with the presence of CD8+ T cells specific for an epitope located between amino acids 320 and 327 of ASP-2. We concluded that a relatively simple formulation consisting of a recombinant protein with a selected portion of ASP-2, alum, and CpG oligodeoxynucleotide 1826 might be used to cross-prime strong CD8+-T-cell-dependent protective immunity against T. cruzi infection.

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Studies on the activity of the hypoxia-inducible-factor hydroxylases using an oxygen consumption assay

Biochemical Journal ◽

10.1042/bj20061151 ◽

2006 ◽

Vol 401 (1) ◽

pp. 227-234 ◽

Cited By ~ 145

Author(s):

Dominic Ehrismann ◽

Emily Flashman ◽

David N. Genn ◽

Nicolas Mathioudakis ◽

Kirsty S. Hewitson ◽

...

Keyword(s):

Oxygen Consumption ◽

Recombinant Protein ◽

Synthetic Peptides ◽

Hypoxia Inducible Factor ◽

Oxygen Sensors ◽

Hydroxylase Activity ◽

Protein Substrates ◽

Physiological Conditions ◽

The Relationship

The activity and levels of the metazoan HIF (hypoxia-inducible factor) are regulated by its hydroxylation, catalysed by 2OG (2-oxoglutarate)- and Fe(II)-dependent dioxygenases. An oxygen consumption assay was developed and used to study the relationship between HIF hydroxylase activity and oxygen concentration for recombinant forms of two human HIF hydroxylases, PHD2 (prolyl hydroxylase domain-containing protein 2) and FIH (factor inhibiting HIF), and compared with two other 2OG-dependent dioxygenases. Although there are caveats on the absolute values, the apparent Km (oxygen) values for PHD2 and FIH were within the range observed for other 2OG oxygenases. Recombinant protein substrates were found to have lower apparent Km (oxygen) values compared with shorter synthetic peptides of HIF. The analyses also suggest that human PHD2 is selective for fragments of the C-terminal over the N-terminal oxygen-dependent degradation domain of HIF-1α. The present results, albeit obtained under non-physiological conditions, imply that the apparent Km (oxygen) values of the HIF hydroxylases enable them to act as oxygen sensors providing their in vivo capacity is appropriately matched to a hydroxylation-sensitive signalling pathway.

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Th-17 cytokines are associated with severity of Trypanosoma cruzi chronic infection in pediatric patients from endemic areas of Mexico

Acta Tropica ◽

10.1016/j.actatropica.2017.11.009 ◽

2018 ◽

Vol 178 ◽

pp. 134-141 ◽

Cited By ~ 11

Author(s):

Mariana De Alba-Alvarado ◽

Paz María Salazar-Schettino ◽

Luis Jiménez-Álvarez ◽

Margarita Cabrera-Bravo ◽

Cecilia García-Sancho ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chronic Infection ◽

Pediatric Patients ◽

Th 17 ◽

Endemic Areas

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The potential risk of enzootic Trypanosoma cruzi transmission inside four training and re-training military battalions (BITER) in Colombia

Parasites & Vectors ◽

10.1186/s13071-021-05018-4 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Omar Cantillo-Barraza ◽

Jeffer Torres ◽

Carolina Hernández ◽

Yanira Romero ◽

Sara Zuluaga ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chagas Disease ◽

Military Personnel ◽

Potential Risk ◽

Epidemiological Surveillance ◽

12S Rrna ◽

Military Population ◽

Endemic Areas ◽

The Military ◽

Feeding Sources

Abstract Background Colombia’s National Army is one of the largest military institutions in the country based on the number of serving members and its presence throughout the country. There have been reports of cases of acute or chronic cases of Chagas disease among active military personnel. These may be the result of military-associated activities performed in jungles and other endemic areas or the consequence of exposure to Trypanosoma cruzi inside military establishments/facilities located in endemic areas. The aim of the present study was to describe the circulation of T. cruzi inside facilities housing four training and re-training battalions [Battalions of Instruction, Training en Re-training (BITERs)] located in municipalities with historical reports of triatomine bugs and Chagas disease cases. An entomological and faunal survey of domestic and sylvatic environments was conducted inside each of these military facilities. Methods Infection in working and stray dogs present in each BITER location was determined using serological and molecular tools, and T. cruzi in mammal and triatomine bug samples was determined by PCR assay. The PCR products of the vertebrate 12S rRNA gene were also obtained and subjected to Sanger sequencing to identify blood-feeding sources. Finally, we performed a geospatial analysis to evaluate the coexistence of infected triatomines and mammals with the military personal inside of each BITER installation. Results In total, 86 specimens were collected: 82 Rhodnius pallescens, two Rhodnius prolixus, one Triatoma dimidiata and one Triatoma maculata. The overall T. cruzi infection rate for R. pallescens and R. prolixus was 56.1 and 100% respectively, while T. dimidiata and T. maculata were not infected. Eight feeding sources were found for the infected triatomines, with opossum and humans being the most frequent sources of feeding (85.7%). Infection was most common in the common opossum Didelphis marsupialis, with infection levels of 77.7%. Sylvatic TcI was the most frequent genotype, found in 80% of triatomines and 75% of D. marsupialis. Of the samples collected from dogs (n = 52), five (9.6%; 95% confidence interval: 3.20–21.03) were seropositive based on two independent tests. Four of these dogs were creole and one was a working dog. The spatial analysis revealed a sympatry between infected vectors and mammals with the military population. Conclusions We have shown a potential risk of spillover of sylvatic T. cruzi transmission to humans by oral and vectorial transmission in two BITER installations in Colombia. The results indicate that installations where 100,000 active military personnel carry out training activities should be prioritized for epidemiological surveillance of Chagas disease. Graphical abstract

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A recombinant protein based on the Trypanosoma cruzi metacyclic trypomastigote 82-kilodalton antigen that induces and effective immune response to acute infection.

Infection and Immunity ◽

10.1128/iai.64.4.1093-1099.1996 ◽

1996 ◽

Vol 64 (4) ◽

pp. 1093-1099 ◽

Cited By ~ 23

Author(s):

R Santori F ◽

G S Paranhos-Bacalla ◽

J Franco DA Silveira ◽

L M Yamauchi ◽

J E Araya ◽

...

Keyword(s):

Immune Response ◽

Recombinant Protein ◽

Trypanosoma Cruzi ◽

Acute Infection ◽

Effective Immune Response ◽

Metacyclic Trypomastigote

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The efficacy of recombinant protein lbk39 for the diagnosis of leishmaniosis in dogs

Parasitology ◽

10.1017/s0031182020001997 ◽

2020 ◽

pp. 1-9

Author(s):

Joyce Carvalho Pereira ◽

Pedro Dos Santos Sousa ◽

Ligia Moraes Barizon De Souza ◽

Aline Kuhn Sbruzzi Pasquali ◽

Michelle Bates ◽

...

Keyword(s):

Recombinant Protein ◽

Amino Acid Identity ◽

Human Infection ◽

Antibody Detection ◽

Leishmania Braziliensis ◽

Total Extract ◽

Elimination Programme ◽

Long Time ◽

Endemic Areas ◽

Visceral Leishmaniosis

Abstract Visceral leishmaniosis is one of the most important zoonotic diseases on the planet and dogs are the main reservoir of canine visceral leishmaniosis (CVL) in endemic areas. They play an important role in human infection because in dogs the disease appears long time after infection, and they can move uncontrollably, contributing to disperse the parasite. To take the decision to treat the animals or for euthanasia, in an elimination programme, in order to reduce the parasitic load, it is necessary to diagnose correctly, having more effective tools. Our group has developed a new recombinant antigen-based kinesin-related gene of Leishmania braziliensis (Lbk39), which shows 59% amino acid identity to the L. infantum homologue. The Lbk39 gene was synthesized, inserted into the pLEXSY-sat2 vector and transfected into L. tarentolae cells by electroporation. The recombinant protein was secreted in the culture with a C-terminal histidine marker, purified, generating a product at 337.68 μg mL−1. A total of 152 sera from dog's endemic and non-endemic areas were used, being 78 positives and 75 negatives. The antigen Lbk39 showed 100% sensitivity and 96.1% specificity. We compared this antigen with other antigens such as total extract of the parasite, TRDPP, and our data indicate that Lbk39 has potential application in the diagnosis of CVL through antibody detection.

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