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1.
PLoS Negl Trop Dis ; 14(8): e0008402, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32797041

RESUMO

A Trypanosoma cruzi Loopamp kit was recently developed as a ready-to-use diagnostic method requiring minimal laboratory facilities. We evaluated its diagnostic accuracy for detection of acute Chagas disease (CD) in different epidemiological and clinical scenarios. In this retrospective study, a convenience series of clinical samples (venous blood treated with EDTA or different stabilizer agents, heel-prick blood in filter paper or cerebrospinal fluid samples (CSF)) from 30 infants born to seropositive mothers (13 with congenital CD and 17 noninfected), four recipients of organs from CD donors, six orally-infected cases after consumption of contaminated guava juice and six CD patients coinfected with HIV at risk of CD reactivation (N = 46 patients, 46 blood samples and 1 CSF sample) were tested by T. cruzi Loopamp kit (Tc LAMP) and standardized quantitative real-time PCR (qPCR). T. cruzi Loopamp accuracy was estimated using the case definition in the different groups as a reference. Cohen's kappa coefficient (κ) was applied to measure the agreement between Tc LAMP (index test) and qPCR (reference test). Sensitivity and specificity of T. cruzi Loopamp kit in blood samples from the pooled clinical groups was 93% (95% CI: 77-99) and 100% (95% CI: 80-100) respectively. The agreement between Tc LAMP and qPCR was almost perfect (κ = 0.92, 95% CI: 0.62-1.00). The T. cruzi Loopamp kit was sensitive and specific for detection of T. cruzi infection. It was carried out from DNA extracted from peripheral blood samples (via frozen EDTA blood, guanidine hydrochloride-EDTA blood, DNAgard blood and dried blood spots), as well as in CSF specimens infected with TcI or TcII/V/VI parasite populations. The T. cruzi Loopamp kit appears potentially useful for rapid detection of T. cruzi infection in congenital, acute and CD reactivation due to HIV infection.


Assuntos
Doença de Chagas/sangue , Doença de Chagas/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Trypanosoma cruzi/isolamento & purificação , Doença de Chagas/líquido cefalorraquidiano , Doença de Chagas/congênito , Coinfecção , DNA de Protozoário/análise , Feminino , Infecções por HIV , Humanos , Lactente , Recém-Nascido , Masculino , Reação em Cadeia da Polimerase em Tempo Real/métodos , Estudos Retrospectivos , Sensibilidade e Especificidade , Transplantados , Trypanosoma cruzi/fisiologia
2.
Cardiovasc Pathol ; 49: 107257, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32674046

RESUMO

BACKGROUND: Chronic Chagas disease (CCHD) associated with Systemic Arterial Hypertension (SAH) is frequently found in areas where the disease is endemic. The pathogenesis of patients with both pathologies (CCHD-SAH) is unsettled. Nitric Oxide (NO) and Kinins are important players in the myocardial inflammation process in experimental CCHD. No previous study has addressed this question in patients with CCHD, particularly in those with CCHD-SAH. Accordingly, this study was undertaken in an attempt to contribute to the understanding of the pathogenesis of patients with CCHD-SAH. METHODS: Thirty-seven patients with a positive serology for Chagas disease were enrolled; 15 had CCHD alone, 22 had CCHD-SAH (abnormal ECG/Doppler echocardiogram plus a systolic blood pressure > 140 mmHg or diastolic blood pressure > 90 mmHg on admission), and 11 had SAH alone. Thirty healthy individuals matched by age and sex served as controls. Plasma High-molecular (Hkg) and low-molecular weight (LKg) kininogens, plasma kallikrein levels (Pkal and Tcal), Kininase II, and plasma NO were measured. RESULTS: HKg and LKg were lower in CCHD-SAH patients in comparison with other groups (P < .0001). Pkal and Tcal were higher in CCHD-SAH patients in comparison with the other groups (P< .0001). Kininase II levels were similar in SAH, CCHD, and CCHD-SAH patients, but lower in comparison with controls (P< .0001). NO levels were similar in CCHD and CCHD-SAH patients, but higher in comparison with SAH patients and controls (P > .0001). CONCLUSION: Such findings suggest increased Kinin and NO activity in patients with CCHD-SAH, thus contributing to the understanding of the pathogenesis of this condition.


Assuntos
Pressão Arterial , Doença de Chagas/sangue , Hipertensão/sangue , Cininas/sangue , Óxido Nítrico/sangue , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Brasil/epidemiologia , Estudos de Casos e Controles , Doença de Chagas/diagnóstico , Doença de Chagas/epidemiologia , Doença de Chagas/fisiopatologia , Feminino , Humanos , Hipertensão/diagnóstico , Hipertensão/epidemiologia , Hipertensão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Regulação para Cima
3.
Mem Inst Oswaldo Cruz ; 115: e190364, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32130371

RESUMO

Oral transmission of Chagas disease has been increasing in Latin American countries. The present study aimed to investigate changes in hepatic function, coagulation factor levels and parasite load in human acute Chagas disease (ACD) secondary to oral Trypanosoma cruzi transmission. Clinical and epidemiological findings of 102 infected individuals attended in the State of Pará from October 2013 to February 2016 were included. The most common symptoms were fever (98%), asthenia (83.3%), face and limb edema (80.4%), headache (74.5%) and myalgia (72.5%). The hepatic enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of 30 ACD patients were higher compared with controls, and this increase was independent of the treatment with benznidazole. Moreover, ACD individuals had higher plasma levels of activated protein C and lower levels of factor VII of the coagulation cascade. Patients with the highest parasite load had also the most increased transaminase levels. Also, ALT and AST were associated moderately (r = 0.429) and strongly (r = 0.595) with parasite load respectively. In conclusion, the present study raises the possibility that a disturbance in coagulation and hepatic function may be linked to human ACD.


Assuntos
Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Doença de Chagas/fisiopatologia , Fator VIIa/análise , Fígado/fisiopatologia , Proteína C/análise , Doença Aguda , Adulto , Biomarcadores/sangue , Brasil/epidemiologia , Estudos de Casos e Controles , Doença de Chagas/sangue , Doença de Chagas/enzimologia , Doença de Chagas/transmissão , Feminino , Humanos , Fígado/enzimologia , Masculino , Pessoa de Meia-Idade , Carga Parasitária , Estudos Prospectivos
4.
Sci Rep ; 10(1): 5294, 2020 03 24.
Artigo em Inglês | MEDLINE | ID: mdl-32210339

RESUMO

The detection of pathogen-specific antibodies remains a cornerstone of clinical diagnostics. Yet, many test exhibit undesirable performance or are completely lacking. Given this, we developed serum epitope repertoire analysis (SERA), a method to rapidly discover conserved, pathogen-specific antigens and their epitopes, and applied it to develop an assay for Chagas disease caused by the protozoan parasite Trypanosoma cruzi. Antibody binding peptide motifs were identified from 28 Chagas repertoires using a bacterial display random 12-mer peptide library and next-generation sequencing (NGS). Thirty-three motifs were selected and mapped to candidate Chagas antigens. In a blinded validation set (n = 72), 30/30 Chagas were positive, 30/30 non-Chagas were negative, and 1/12 Leishmania sp. was positive. After unblinding, a Leishmania cross-reactive epitope was identified and removed from the panel. The Chagas assay exhibited 100% sensitivity (30/30) and specificity (90/90) in a second blinded validation set including individuals with other parasitic infections. Amongst additional epitope repertoires with unknown Chagas serostatus, assay specificity was 99.8% (998/1000). Thus, the Chagas assay achieved a combined sensitivity and specificity equivalent or superior to diagnostic algorithms that rely on three separate tests to achieve high specificity. NGS-based serology via SERA provides an effective approach to discover antigenic epitopes and develop high performance multiplex serological assays.


Assuntos
Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/imunologia , Doença de Chagas/sangue , Doença de Chagas/diagnóstico , Epitopos/imunologia , Trypanosoma cruzi/imunologia , Adulto , Anticorpos Antiprotozoários/sangue , Doença de Chagas/imunologia , Doença de Chagas/parasitologia , Feminino , Humanos , Masculino , Biblioteca de Peptídeos
5.
PLoS One ; 15(1): e0227828, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31951634

RESUMO

Trypanosoma cruzi, the protozoan agent of Chagas disease in the Americas, is comprised of six genetic lineages (TcI-TcVI) and a possible seventh (TcBat, related to TcI). Identification of T. cruzi lineages infecting reservoir mammalian species is fundamental to resolving transmission cycles. However, this is hindered by the limited sensitivity and technical complexity of parasite isolation and genotyping. An alternative approach is serology using T. cruzi lineage-specific epitopes, such as those of the trypomastigote small surface antigen (TSSA). For surveillance of T. cruzi lineage infections in mammal species from diverse Brazilian regions, we apply a novel rapid diagnostic test (RDT, Chagas Sero K-SeT), which incorporates the TSSA peptide epitope specific to TcII/V/VI (TSSApep-II/V/VI) and Protein G detection of antibodies. Chagas Sero K-SeT RDT results with sera from experimentally infected mice, from tamarin primates (Leontopithecus spp.) and from canines (Canis familiaris) were concordant with corresponding TSSApep-II/V/VI ELISAs. The Chagas Sero K-Set detected TcII/V/VI infections in Leontopithecus spp. from the Atlantic forest (n = 46), in C. familiaris (n = 16) and Thrichomys laurentius (n = 2) from Caatinga biome and Chiroptera (n = 1) from Acre, Amazonia. The Chagas Sero K-SeT RDT is directly applicable to TcII/V/VI-specific serological surveillance of T. cruzi infection in several different mammalian Orders. It can replace ELISAs and provides efficient, point-of-sampling, low-cost detection of TcII/V/VI infections, with at least equivalent sensitivity, although some mammals may be difficult to trap, and, not unexpectedly, Chagas Sero K-SeT could not recognise feline IgG. Knowledge of sylvatic hosts of T. cruzi can be expanded, new reservoir species discovered, and the ecology of transmission cycles clarified, particularly with adaptation to further mammalian Orders.


Assuntos
Doença de Chagas/veterinária , Trypanosoma cruzi/isolamento & purificação , Animais , Antígenos de Protozoários/sangue , Antígenos de Protozoários/imunologia , Brasil/epidemiologia , Gatos , Doença de Chagas/sangue , Doença de Chagas/diagnóstico , Testes Diagnósticos de Rotina , Cães , Ensaio de Imunoadsorção Enzimática , Humanos , Camundongos , Trypanosoma cruzi/imunologia
7.
Am J Trop Med Hyg ; 102(1): 159-163, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31701864

RESUMO

It is known that the immunoregulatory networks in human Chagas disease play a key role in parasitemia control during the acute phase. However, little is known regarding the control of parasitemia during the chronic phase. The aim of the study was to describe the serum cytokine profile of Trypanosoma cruzi chronically infected patients and to evaluate its relationship with the presence or absence of parasitemia in peripheral blood. This is a prospective observational study where adult Chagas disease patients were included. Patients previously treated for Chagas disease, pregnant women, and immunosuppressed patients were excluded. Demographic and clinical information was collected, and T. cruzi real-time polymerase chain reaction (RT-PCR) and serum cytokine profile were determined in peripheral blood. Forty-five patients were included. Trypanosoma cruzi RT-PCR in peripheral blood resulted positive in 19 (42.2%) patients. No differences in the serum cytokine profile were found depending on cardiac or digestive involvement. However, patients with positive T. cruzi RT-PCR had a higher median concentration of IL-10 and IL-1beta and a lower median concentration of IL-8 than those with negative T. cruzi PCR. These results reinforce the key role that this anti-inflammatory cytokine (IL-10) plays in parasitemia control.


Assuntos
Doença de Chagas/sangue , Interleucina-10/sangue , Parasitemia/sangue , Adulto , Doença de Chagas/patologia , Doença Crônica , DNA de Protozoário , Feminino , Humanos , Interleucina-10/metabolismo , Interleucina-1beta/sangue , Interleucina-8/sangue , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Adulto Jovem
8.
PLoS One ; 14(12): e0225588, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31841511

RESUMO

Macrophages can reprogram their metabolism in response to the surrounding stimuli, which affects their capacity to kill intracellular pathogens. We have investigated the metabolic and immune status of human macrophages after infection with the intracellular trypanosomatid parasites Leishmania donovani, L. amazonensis and T. cruzi and their capacity to respond to a classical polarizing stimulus (LPS and IFN-γ). We found that macrophages infected with Leishmania preferentially upregulate oxidative phosphorylation, which could be contributed by both host cell and parasite, while T. cruzi infection did not significantly increase glycolysis or oxidative phosphorylation. Leishmania and T. cruzi infect macrophages without triggering a strong inflammatory cytokine response, but infection does not prevent a potent response to LPS and IFN-γ. Infection appears to prime macrophages, since the cytokine response to activation with LPS and IFN-γ is more intense in infected macrophages compared to uninfected ones. Metabolic polarization in macrophages can influence infection and immune evasion of these parasites since preventing macrophage cytokine responses would help parasites to establish a persistent infection. However, macrophages remain responsive to classical inflammatory stimuli and could still trigger inflammatory cytokine secretion by macrophages.


Assuntos
Doença de Chagas/imunologia , Citocinas/metabolismo , Leishmaniose/imunologia , Ativação de Macrófagos , Macrófagos/imunologia , Células 3T3 , Animais , Células Cultivadas , Doença de Chagas/sangue , Doença de Chagas/parasitologia , Citocinas/imunologia , Voluntários Saudáveis , Humanos , Leishmania donovani/imunologia , Leishmania donovani/isolamento & purificação , Leishmania mexicana/imunologia , Leishmania mexicana/isolamento & purificação , Leishmaniose/sangue , Leishmaniose/parasitologia , Macrófagos/metabolismo , Metaboloma/imunologia , Camundongos , Fosforilação Oxidativa , Cultura Primária de Células , Trypanosoma cruzi/imunologia , Trypanosoma cruzi/isolamento & purificação , Regulação para Cima
9.
Mediators Inflamm ; 2019: 5091630, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31772504

RESUMO

Infection with the protozoan Trypanosoma cruzi causes Chagas disease and consequently leads to severe inflammatory heart condition; however, the mechanisms driving this inflammatory response have not been completely elucidated. Nitric oxide (NO) is a key mediator of parasite killing in T. cruzi-infected mice, and previous studies have suggested that leukotrienes (LTs) essentially regulate the NO activity in the heart. We used infected 5-lipoxygenase-deficient mice (5-LO-/-) to explore the participation of nitric oxide synthase isoforms, inducible (iNOS) and constitutive (cNOS), in heart injury, cytokine profile, and oxidative stress during the early stage of T. cruzi infection. Our evidence suggests that the cNOS of the host is involved in the resistance of 5-LO-/- mice during T. cruzi infection. iNOS inhibition generated a remarkable increase in T. cruzi infection in the blood and heart of mice, whereas cNOS inhibition reduced cardiac parasitism (amastigote nests). Furthermore, this inhibition associates with a higher IFN-γ production and lower lipid peroxidation status. These data provide a better understanding about the influence of NO-interfering therapies for the inflammatory response toward T. cruzi infection.


Assuntos
Araquidonato 5-Lipoxigenase/sangue , Doença de Chagas/sangue , Doença de Chagas/enzimologia , Animais , Antioxidantes/metabolismo , Citocinas/sangue , Camundongos , Camundongos Knockout , Óxido Nítrico/sangue , Óxido Nítrico Sintase Tipo II/sangue , Trypanosoma cruzi/patogenicidade
10.
PLoS Negl Trop Dis ; 13(9): e0007715, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31553732

RESUMO

BACKGROUND: The National Program for Chagas disease was implemented in Bolivia in 2006, and it greatly decreased the number of infections through vector control. Subsequently, a treatment regimen of benznidazole (BNZ) was started in seropositive school-age children living in certified vector control areas. METHODS AND FINDINGS: We conducted a 12-month follow-up study and seven blood samples were taken during and after the treatment. Serology, conventional diagnostic PCR (cPCR) and quantitative Real-time PCR (qPCR) were performed. Plasma Th1/Th2/Th17 cytokines levels were also determined. Approximately 73 of 103 seropositive children complied with BNZ, with three interruptions due to side effects. To evaluate each individual's treatment efficacy, the cPCR and qPCR values during the final 6 months of the follow-up period were observed. Among 57 children who completed follow-up, 6 individuals (11%) showed both cPCR(+) and qPCR(+) (non reactive), 24 (42%) cPCR(-) but qPCR(+) (ambiguous) and 27 (47%) cPCR(-) and qPCR(-) (reactive). Within 14 Th1/Th2/Th17 cytokines, IL-17A showed significantly higher levels in seropositive children before the treatment compared to age-matched seronegative children and significantly decreased to the normal level one-year after. Moreover, throughout the follow-up study, IL-17A levels were positively co-related to parasite counts detected by qPCR. At the 12 months' time point, IL-17A levels of non-reactive subjects were significantly higher than either those of reactive or ambiguous subjects suggesting that IL-17A might be useful to determine the reactivity to BNZ treatment. CONCLUSIONS: Plasma levels of IL-17A might be a bio-marker for detecting persistent infection of T. cruzi and its chronic inflammation.


Assuntos
Doença de Chagas/tratamento farmacológico , Interleucina-17/sangue , Nitroimidazóis/uso terapêutico , Resultado do Tratamento , Adolescente , Biomarcadores/sangue , Bolívia , Doença de Chagas/sangue , Criança , Pré-Escolar , Citocinas , Feminino , Seguimentos , Humanos , Masculino , Nitroimidazóis/sangue , Reação em Cadeia da Polimerase/métodos , Tripanossomicidas/sangue , Tripanossomicidas/uso terapêutico , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/isolamento & purificação
11.
JCI Insight ; 4(18)2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31479429

RESUMO

Chagas disease is a lifelong pathology resulting from Trypanosoma cruzi infection. It represents one of the most frequent causes of heart failure and sudden death in Latin America. Herein, we provide evidence that aerobic glycolytic pathway activation in monocytes drives nitric oxide (NO) production, triggering tyrosine nitration (TN) on CD8+ T cells and dysfunction in patients with chronic Chagas disease. Monocytes from patients exhibited a higher frequency of hypoxia-inducible factor 1α and increased expression of its target genes/proteins. Nonclassical monocytes are expanded in patients' peripheral blood and represent an important source of NO. Monocytes entail CD8+ T cell surface nitration because both the frequency of nonclassical monocytes and that of NO-producing monocytes positively correlated with the percentage of TN+ lymphocytes. Inhibition of glycolysis in in vitro-infected peripheral blood mononuclear cells decreased the inflammatory properties of monocytes/macrophages, diminishing the frequency of IL-1ß- and NO-producing cells. In agreement, glycolysis inhibition reduced the percentage of TN+CD8+ T cells, improving their functionality. Altogether, these results clearly show that glycolysis governs oxidative stress on monocytes and modulates monocyte-T cell interplay in human chronic Chagas disease. Understanding the pathological immune mechanisms that sustain an inflammatory environment in human pathology is key to designing improved therapies.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Comunicação Celular/imunologia , Doença de Chagas/imunologia , Glicólise/imunologia , Monócitos/metabolismo , Trypanosoma cruzi/imunologia , Adulto , Animais , Linfócitos T CD8-Positivos/efeitos dos fármacos , Estudos de Casos e Controles , Comunicação Celular/efeitos dos fármacos , Doença de Chagas/sangue , Doença de Chagas/tratamento farmacológico , Chlorocebus aethiops , Técnicas de Cocultura , Feminino , Glicólise/efeitos dos fármacos , Humanos , Ativação Linfocitária/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/imunologia , Cultura Primária de Células , Proteínas de Protozoários/imunologia , Tirosina/metabolismo , Células Vero , Adulto Jovem
12.
Molecules ; 24(17)2019 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-31480397

RESUMO

Trypanosoma cruzi, the etiological agent of Chagas disease, is dependent on proline for a variety of processes, such as energy metabolism, host cell invasion, differentiation, and resistance to osmotic, metabolic, and oxidative stress. On this basis, we investigated a possible relationship between prolinemia and severity of T. cruzi infection in chronic patients, as reported here. The study population consisted of 112 subjects, separated into 83 chronically T. cruzi-infected patients and 29 age-matched healthy volunteers (control) of both sexes, recruited at the Chagas Disease Service from the Department of Cardiology, Hospital Provincial del Centenario de Rosario (Rosario, Argentina). Chagasic patients were separated into three groups: chronic asymptomatic, mild/moderate, and severe chronic chagasic cardiomyopathy (CCC) subjects. We observed a significant decrease of 11.7% in prolinemia in chagasic patients when compared to controls. Further analysis within the three groups of chagasic patients also revealed a statistically significant decrease of prolinemia in severe CCC patients compared to controls, showing a relative difference of 13.6% in proline concentrations. These data point to the possibility that collagen-which participates in the healing process of cardiac tissue-and proline metabolism in the myocardium could constitute new factors affecting the evolution of Chagas disease.


Assuntos
Doença de Chagas/sangue , Doença de Chagas/patologia , Prolina/sangue , Adolescente , Adulto , Idoso , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
13.
Am J Trop Med Hyg ; 101(5): 1135-1138, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31516110

RESUMO

The discovery and characterization of novel parasite antigens to improve the diagnosis of Trypanosoma cruzi by serological methods and for accurate and rapid follow-up of treatment efficiency are still needed. TcTASV is a T. cruzi-specific multigene family, whose products are expressed on the parasite stages present in the vertebrate host. In a previous work, a mix of antigens from subfamilies TcTASV-A and TcTASV-C (Mix A + C) was sensitive and specific to identify dogs with active infection of high epidemiological relevance. Here, TcTASV-A and TcTASV-C were assayed separately as well as together (Mix A + C) in an ELISA format on human samples. The Mix A + C presented moderate sensitivity (78%) but high diagnostic accuracy with a 100% of specificity, evaluated on healthy, leishmaniasic, and Strongyloides stercoralis infected patients. Moreover, antibody levels of pediatric patients showed-2 years posttreatment-diminished reactivity against the Mix A + C (P < 0.0001), pointing TcTASV antigens as promising tools for treatment follow-up.


Assuntos
Antígenos de Protozoários/sangue , Antiprotozoários/uso terapêutico , Doença de Chagas/sangue , Doença de Chagas/tratamento farmacológico , Trypanosoma cruzi/imunologia , Anticorpos Antiprotozoários/imunologia , Biomarcadores/sangue , Criança , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Sensibilidade e Especificidade
14.
Rev Soc Bras Med Trop ; 52: e20190133, 2019 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-31508781

RESUMO

INTRODUCTION: Chagas disease (CD) is an important public health problem in Brazil and worldwide. Aging and obesity are important matters in patients with CD, as is hypovitaminosis D3, which can decrease the quality of life of these patients. Immunomodulation mediated by vitamin D3, especially the production of antimicrobial peptides such as cathelicidin LL-37, might be related to the severity and symptoms of CD. This study aimed to determine the serum levels of vitamin D and LL-37 and VDR gene polymorphisms in patients with chronic CD. METHODS: This study included male patients with cardiac and indeterminate clinical forms of CD. Clinical, anthropometric, and blood parameters were obtained. Serum levels of 25(OH)D3 and LL-37 were determined by chemiluminescence and enzyme-linked immunosorbent assay respectively. Fok (rs731236), Bsm (rs1544410), Apa (rs7975232), and Taq (rs731236) polymorphisms of the VDR gene were investigated by PCR-RFLP. RESULTS: Sixty-four patients were included in the study: 18 of the cardiac form and 46 of the indeterminate form. No differences in age, ethnicity, BMI, arterial hypertension, diabetes mellitus, or dyslipidemias were observed between groups. However, the serum levels of 25(OH)D3, but not of LL-37, were lower in the cardiac form group. The association among polymorphisms, vitamin D, and clinical form was not significant. CONCLUSIONS: Decreased levels of vitamin D suggest an association with the cardiac form of CD. Studies investigating the roles of vitamin D and LL-37 in the immune response and their associations with VDR polymorphisms and disease susceptibility are necessary.


Assuntos
Peptídeos Catiônicos Antimicrobianos/sangue , Doença de Chagas/sangue , Doença de Chagas/genética , Colecalciferol/sangue , Polimorfismo Genético/genética , Receptores de Calcitriol/genética , Biomarcadores/sangue , Humanos , Masculino , Pessoa de Meia-Idade
15.
Korean J Parasitol ; 57(4): 435-437, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31533412

RESUMO

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.


Assuntos
Anticorpos Antiprotozoários/sangue , Doença de Chagas/diagnóstico , Proteínas Ribossômicas/imunologia , Trypanosoma cruzi/imunologia , Western Blotting , Brasil , Doença de Chagas/sangue , Doença de Chagas/imunologia , Doenças Endêmicas , Humanos , Proteínas Recombinantes/imunologia
16.
ACS Infect Dis ; 5(11): 1813-1819, 2019 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-31538468

RESUMO

trans-Sialidase and cruzipain are important virulence factors from Trypanosoma cruzi, the etiological agent of Chagas disease, that have highly antigenic domains in their structure and were reported as potential tools for diagnosis of the illness. The aim of the present study is to assess the possibility of using cruzipain and the catalytic domain of trans-sialidase in a Surface Plasmon Resonance-based immunosensor for the diagnosis of chronic Chagas disease. Immunoassays carried out with canine sera verified that cruzipain allows the detection of anti-Trypanosoma cruzi antibodies whereas recombinant trans-sialidase did not yield specific detections, due to the high dilutions of serum used in the immunoassays that hinder the possibility to sense the specific low titer antibodies. The developed cruzipain-based biosensor, whose price per assay is comparable to a commercial enzyme-linked immunosorbent assay (ELISA), was successfully applied for the rapid quantification of specific antibodies against Trypanosoma cruzi in fresh human sera showing an excellent agreement with ELISA.


Assuntos
Anticorpos Antiprotozoários/sangue , Doença de Chagas/diagnóstico , Doença de Chagas/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Trypanosoma cruzi/isolamento & purificação , Animais , Doença de Chagas/sangue , Doença de Chagas/parasitologia , Cisteína Endopeptidases/análise , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/imunologia , Doenças do Cão/sangue , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Glicoproteínas/análise , Glicoproteínas/genética , Glicoproteínas/imunologia , Humanos , Neuraminidase/análise , Neuraminidase/genética , Neuraminidase/imunologia , Proteínas de Protozoários/análise , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Trypanosoma cruzi/genética , Trypanosoma cruzi/imunologia , Fatores de Virulência/sangue , Fatores de Virulência/genética , Fatores de Virulência/imunologia
17.
Acta Trop ; 200: 105167, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31513762

RESUMO

It is not currently known which individuals with chronic Chagas disease (ChD) will develop cardiopathy in a determined period and which will be maintained asymptomatic with normal routine laboratory tests all their lives. The parasite burden is a factor that could explain this different evolution. The objective of this study was to quantify Trypanosoma cruzi burden by real-time PCR in blood (qPCR-B) and dejections of triatomines fed by xenodiagnosis (qPCR-XD) in 90 individuals with chronic ChD untreated, classified according to XD results and the presence or absence of cardiopathy. All individuals came from hyperendemic areas of Chile and participated in the study under Informed Consent. The standard qPCR curves for qPCR-B and qPCR-XD were elaborated with a mixture of known concentrations of T. cruzi strains, performing DNA serial dilutions (1/10) with a dynamic range between 105 and 10-1 parasite equivalents/mL. The TaqManⓇ detection system was applied in a Stratagene Mx3000P thermocycler (Agilent Technologies, USA) with cruzi 1 and cruzi 2 satellite primers. 22.2% and 15.6% of cases with cardiopathy or without cardiopathy were XD positive. There was no significant difference between the groups. The positivity of qPCR-B and qPCR-XD in the positive XD group was 82.35% and 100%, respectively, while in the negative XD group was 55.26% and 42.10%, respectively. A superior qPCR value in chronic ChD patients with and without cardiopathy was determined for qPCR in cases with positive XD and positive qPCR-XD. The receiver operating characteristic (ROC) curve analyses show better accuracy for detecting parasite burden (area under the curve, AUC) for qPCR-XD in comparison to qPCR-B. That is to say, major performance in DNA samples obtained of positive XD (gold standard for viable T. cruzi) detected and quantified by qPCR-XD. A high percentage of cases with XD and qPCR-XD positive (80-100%) have result concordant with qPCR-B. In absence of XD, future challenges are especially related to the low parasitic load of chronic ChD patients treated with trypanocidal drugs and post-therapy parasitological evaluations by qPCR-B. Finally, no statistically significant differences were found between presence or absence of cardiopathy and XD, qPCR-B or qPCR-XD.


Assuntos
Doença de Chagas/complicações , Doença de Chagas/parasitologia , Cardiopatias/etiologia , Carga Parasitária , Triatoma/parasitologia , Trypanosoma cruzi/isolamento & purificação , Xenodiagnóstico/métodos , Adulto , Fatores Etários , Idoso , Animais , Doença de Chagas/sangue , Doença de Chagas/epidemiologia , Chile/epidemiologia , Doença Crônica/epidemiologia , Testes Diagnósticos de Rotina , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real/métodos , Tripanossomicidas , Trypanosoma cruzi/genética
18.
Front Immunol ; 10: 1671, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31379862

RESUMO

Monocytes are classified according to their CD14 and CD16 expression into classical (reparative), intermediate (inflammatory), and non-classical. This study assessed the frequency of monocyte and the relationship between monocyte subset percentages and the levels of blood cytokines in Colombian chagasic patients with different clinical forms. This study included chagasic patients in different clinical stages: indeterminate (IND) n = 14, chronic chagasic cardiomyopathy (CCC) n = 14, and heart transplant chagasic (HTCC) n = 9; controls with non-chagasic cardiopathy (NCC) n = 15, and healthy individuals (HI) n = 15. Peripheral blood mononuclear cells (PBMCs) were isolated, labeled for CD14, CD16, and HLA-DR, and analyzed by flow cytometry. Cytokines were measured with a bead-based immunoassay. Percentages of total CD14+ CD16+ and CD14+ HLA-DR+ monocytes were higher in patients with heart involvement (CCC, HTCC, and NCC) than controls. Percentages of intermediate monocytes increased in symptomatic chagasic patients (CCC and HTCC) compared to asymptomatic chagasic patients (IND) and controls (HI). Asymptomatic chagasic patients (IND) had higher percentages of classical monocytes, an increased production of CCL17 chemokine compared to chagasic symptomatic patients (CCC), and their levels of CCL17 was positively correlated with the percentage of classical monocyte subset. In CCC, the percentages of intermediate and classical monocytes were positively correlated with IL-6 levels, which were higher in this group compared to HI, and negatively with IL-12p40 concentration, respectively. Remarkably, there also was an important increased of classical monocytes frequency in three chronic chagasic patients who underwent cardiac transplant, of which one received anti-parasitic treatment. Our findings suggest that cardiac chagasic patients have an increased percentage of inflammatory monocytes and produce more IL-6, a biomarker of heart failure and left ventricular dysfunction, whereas asymptomatic chagasic individuals present a higher percentage of reparative monocytes and CCL17.


Assuntos
Doença de Chagas/imunologia , Citocinas/imunologia , Monócitos/imunologia , Adulto , Idoso , Doença de Chagas/sangue , Citocinas/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença
19.
Rev Soc Bras Med Trop ; 52: e20190146, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31390443

RESUMO

INTRODUCTION: Chagas disease (CD) affects 5.7-7.0 million individuals worldwide, and its prevalence reached 25.1% in the state of Bahia, Brazil. There is an association between the prevalence of CD, the socioeconomic status of the population, and the risk of re-emergence due to non-vectorial transmission, such as blood transfusion. This study determined the seroprevalence of T. cruzi infection among blood donors in the state of Bahia, located in northeastern Brazil, and their epidemiological profile during a 10-year period. METHODS: We performed a descriptive cross-sectional study involving a database review. Data were collected from patients with non-negative results for T. cruzi infection during a 10-year period. RESULTS: A total of 3,084 (0.62%) samples were non-negative for T. cruzi infection in an initial serological screening, and 810 (0.16%) samples were non-negative in the second screening. The correlation between infection and age (30 years or older) and between infection and lower educational level (12 years or less) in the first and second screening was statistically significant. The seroprevalence of T. cruzi infection was higher in men in the first screening. In addition, 99.52% of the municipalities of Bahia had at least one case of CD. Livramento de Nossa Senhora and Salvador presented the highest disease prevalence and recurrence, respectively. CONCLUSIONS: The seroprevalence of T. cruzi infection in these populations was lower than that found in other studies in Brazil but was comparatively higher in densely-populated areas. The demographic characteristics of our population agreed with previous studies.


Assuntos
Doadores de Sangue/estatística & dados numéricos , Doença de Chagas/epidemiologia , Trypanosoma cruzi/isolamento & purificação , Distribuição por Idade , Anticorpos Antiprotozoários/sangue , Brasil/epidemiologia , Doença de Chagas/sangue , Doença de Chagas/transmissão , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Programas de Rastreamento/estatística & dados numéricos , Prevalência , Estudos Soroepidemiológicos , Distribuição por Sexo , Fatores Socioeconômicos
20.
Int J Infect Dis ; 87: 100-108, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31357062

RESUMO

OBJECTIVE: Chagas disease affects over six million people, but less than 1% are diagnosed and treated. Complicated diagnostic processes are a major barrier. Colombia's previous diagnostic algorithm, using in-house tests, was difficult to scale up, creating significant access barriers for patients. A new algorithm using commercially manufactured immunoassays would potentially improve access, but these tests' performance in Colombian patients with Chagas disease is not well known. METHODS: We assessed seven commercially available assays. Samples (n=501), 93.8% originating from Colombia, were characterized as positive or negative based on standard procedure at the National Reference Laboratory. Performance characteristics were calculated for individual assays and hypothetical test pairings, then compared to the existing algorithm. RESULTS: Five of seven assays exhibited sensitivity >98% while six showed specificity >97%. A total antigen ELISA paired with a recombinant assay provided similar performance to the current diagnostic process. Six of six assays tested proved capable of detecting different Trypanosoma cruzi genetic lineages. CONCLUSIONS: The study indicated that several commercial assays accurately detect T. cruzi infection in Colombian patients. A simplified testing process with two commercial assays could perform comparably to the previous process, reducing cost and accessibility barriers and facilitating national scale-up.


Assuntos
Doença de Chagas/diagnóstico , Imunoensaio/métodos , Anticorpos Antiprotozoários/sangue , Doença de Chagas/sangue , Doença de Chagas/parasitologia , Colômbia , Humanos , Sensibilidade e Especificidade , Trypanosoma cruzi/genética , Trypanosoma cruzi/fisiologia
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