scholarly journals Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (Viannia) braziliensis

1996 ◽  
Vol 38 (3) ◽  
pp. 177-185 ◽  
Author(s):  
Ana de Cássia Vexenat ◽  
Jaime M. Santana ◽  
Antonio R.L. Teixeira
Keyword(s):  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Cross Reactivity ◽  
Antigenic Determinants ◽  
Leishmania Braziliensis ◽  
Leishmania Chagasi ◽  
Mucocutaneous Leishmaniasis ◽  
Kala Azar ◽  
Homologous Antigen ◽  
Positive Results

We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100% positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100% positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100% positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95% of Kala-azar and 35% of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15% of Chagas sera agglutinated the parasite in dilutions 1:16 and below. Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.

scholarly journals Use of elisa employing homologous and heterologous antigens for the detection of IgG and subclasses (IgG1 and IgG2) in the diagnosis of Canine visceral leishmaniasis

2011 ◽  
Vol 53 (5) ◽  
pp. 283-289 ◽  
Author(s):  
Flávia Coelho Ribeiro ◽  
Armando de O. Schubach ◽  
Eliame Mouta-Confort ◽  
Tânia M.V. Pacheco ◽  
Maria de Fátima Madeira ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Large Scale ◽  
Cross Reactivity ◽  
Leishmania Braziliensis ◽  
Leishmania Chagasi ◽  
Serum Samples ◽  
Canine Ehrlichiosis ◽  
Control Serum ◽  
Homologous Antigen ◽  
Heterologous Antigens

Indirect immunofluorescence is the method recommended for the diagnosis of visceral leishmanisis in dogs, however, the accuracy of this technique is low and its use on a large scale is limited. Since ELISA does not present these limitations, this technique might be an option for the detection of IgG or specific IgG1 and IgG2 subclasses. Canine ehrlichiosis is an important differential diagnosis of American Visceral Leishmaniasis (AVL). The present study compared ELISA using Leishmania chagasi and Leishmania braziliensis antigen for the detection of anti-Leishmania IgG and subclasses in serum samples from 37 dogs naturally infected with L. chagasi (AVL) and in samples from four dogs co-infected with L. braziliensis and L. chagasi (CI). The occurrence of cross-reactivity was investigated in control serum samples of 17 healthy dogs (HC) and 35 infected with Ehrlichia canis (EC). The mean optical density obtained for the detection of IgG was significantly higher when L. chagasi antigen was used, and was also higher in subgroup VLs (symptomatic) compared to subgroup Vla (asymptomatic). The correlation between IgG and IgG1 was low. The present results suggest that IgG ELISA using homologous antigen yields the best results, permitting the diagnosis of asymptomatic L. chagasi infection and the discrimination between cases of AVL and ehrlichiosis in dogs.

scholarly journals Comparison on the performance of Leishmania major-like and Leishmania braziliensis braziliensis as antigen for new world leishmaniasis IgG-immunofluorescence test

1991 ◽  
Vol 33 (6) ◽  
pp. 503-508 ◽  
Author(s):  
Maria Carolina Soares Guimarães ◽  
Beatriz J. Celeste ◽  
Edelma María Corrales L. ◽  
Carlos M.F. Antunes
Keyword(s):  
Visceral Leishmaniasis ◽  
Chagas Disease ◽  
Leishmania Major ◽  
Cross Reactivity ◽  
Leishmania Braziliensis ◽  
Nuclear Factor ◽  
Mucocutaneous Leishmaniasis ◽  
Immunofluorescent Test ◽  
Normal Controls ◽  
Better Than

The performance of an antigen of L. major-like promastigotes for the serological diagnosis of mucocutaneous leishmaniasis in the IgG-immunofluorescent test was compared to that of an antigen of L. braziliensis braziliensis. Each antigen was used to test two hundred and twenty-four sera of etiologies such as mucocutaneous leishmaniasis, deep mycoses, toxoplasmosis, malaria, Chagas' disease, visceral leishmaniasis, anti-nuclear factor, schistosomaiasis, rheumatoid factor and normal controls. Agreement between responses to each antigen was high: 77.2% of leishmaniases sera agreed on a positive or a negative result to both antigens and 91.1 % of control sera. Cross reactivity was restricted to Chagas' disease sera, visceral leishmaniasis, anti-nuclear factor and paracoccidiodomycosis. The quantitative response of leishmaniasis and Chagas' disease sera to both antigens was evaluated by a linear regression; although the y-intercept and the slope were different for each antigen, neither was better than the other in the disclosure of anti-Leishmania antibodies. In the case of Chagas' disease sera the L. major-like antigen was better than L. b. braziliensis' to disclose cross-reacting antibodies.

scholarly journals Trypanosoma cruzi: identification of specific epimastigote antigens by human immune sera

1989 ◽  
Vol 84 (3) ◽  
pp. 309-314 ◽  
Author(s):  
M. G. Morgado ◽  
J. Ivo-dos-Santos ◽  
R. T. Pinho ◽  
E. Argüelles ◽  
J. M. Rezende ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Trypanosoma Cruzi ◽  
Western Blot ◽  
Chagas Disease ◽  
Cross Reactivity ◽  
Cross Reaction ◽  
The Other ◽  
Molecular Weights ◽  
Human Visceral Leishmaniasis ◽  
Human Immune

Soluble antigens from epimastigotes of Trypanosoma cruzi were analyzed by western blot in terms of their reactivity with sera from patients with Chagas' disease. In addition, sera from patients with visceral (AVL) and tegumentar leishmaniasis (ATL) were also tested in order to identify cross-reactivities with Trypanosoma cruzy antigens. Twenty eight polypeptides with molecular weights ranging from 14 kDa to 113 kDa were identified with sera from Chagas' disease patients. An extensive cross-reactivity was observed when sera from human visceral leishmaniasis were used, while only a slight cross-reaction was observed with sera from tegumentar leishmaniasis. On the other hand, 10 polypeptidesspecifically reacting with sera from Chagas' disease patients were identified. Among them, the antigens with molecular weights of 46 kDa and 25 kDa reacted with all sera teste and may be good candidates for specific immunodiagnosis of Chagas' disease.

Lipid biosynthesis pathways as chemotherapeutic targets in kinetoplastid parasites

Parasitology ◽  
1997 ◽  
Vol 114 (7) ◽  
pp. 91-99 ◽  
Author(s):  
J. A. URBINA
Keyword(s):  
Trypanosoma Cruzi ◽  
Recent Work ◽  
Chagas Disease ◽  
Pneumocystis Carinii ◽  
Lipid Biosynthesis ◽  
Sterol Biosynthesis ◽  
Leishmania Braziliensis ◽  
Biosynthesis Inhibitors

Inhibitors of sterol and phospholipid biosynthesis in kinetoplastid parasites such as Trypanosoma cruzi, the causative agent of Chagas' disease, and different species of Leishmania have potent and selective activity as chemotherapeutic agents in vitro and in vivo. Recent work with the sterol C14α-demethylase inhibitor D0870, a bis triazole derivative, showed that this compound is capable of inducing radical parasitological cure in murine models of both acute and chronic Chagas' disease. Other inhibitors of this type, such as SCH 56592, have also shown curative, rather than suppressive, activity against T. cruzi in these models. Leishmania species have different susceptibilities to sterol biosynthesis inhibitors, both in vitro and in vivo. Leishmania braziliensis promastigotes, naturally resistant to C14α-demethylase inhibitors such as ketoconazole and D0870, were susceptible to these drugs when used in combination with the squalene epoxidase inhibitor terbinafine. Inhibitors of Δ24(25) sterol methyl transferase have been shown to act as potent antiproliferative agents against Trypanosoma cruzi, both in vitro and in vivo. New inhibitors of this type which show enhanced activity and novel mechanisms of action have been synthesized. Recent work has also demonstrated that this type of enzyme inhibitors can block sterol biosynthesis and cell proliferation in Pneumocystis carinii, a fungal pathogen which had previously been found resistant to other sterol biosynthesis inhibitors. Ajoene, an antiplatelet compound derived from garlic, was shown to have potent antiproliferative activity against epimastigotes and amastigotes of Trypanosoma cruzi in vitro; this activity was associated with a significant alteration of the phospholipid composition of the cells with no significant effects on the sterol content. In addition, alkyllsophospholipids such as ilmofosine, miltefosine and edelfosine have been shown to block the proliferation of T. cruzi and Leishmania and alter both the phospholipid and sterol composition. These results indicate the potential of lipid biosynthesis inhibitors as useful therapeutic agents in the treatment of leishmaniasis and Chagas' disease.

TESA-blot for the diagnosis of Chagas disease in dogs from co-endemic regions for Trypanosoma cruzi, Trypanosoma evansi and Leishmania chagasi

Acta Tropica ◽  
2009 ◽  
Vol 111 (1) ◽  
pp. 15-20 ◽  
Author(s):  
E.S. Umezawa ◽  
A.I. Souza ◽  
V. Pinedo-Cancino ◽  
M. Marcondes ◽  
A. Marcili ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Trypanosoma Evansi ◽  
Leishmania Chagasi

scholarly journals Mapping Antigenic Motifs in the Trypomastigote Small Surface Antigen from Trypanosoma cruzi

2015 ◽  
Vol 22 (3) ◽  
pp. 304-312 ◽  
Author(s):  
Virginia Balouz ◽  
María de los Milagros Cámara ◽  
Gaspar E. Cánepa ◽  
Santiago J. Carmona ◽  
Romina Volcovich ◽  
...  
Keyword(s):  
Amino Acid ◽  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Surface Antigen ◽  
Humoral Response ◽  
Antigenic Structure ◽  
Antigenic Determinants ◽  
Small Surface ◽  
Content Type ◽  
Surface Receptors

ABSTRACTThe trypomastigote small surface antigen (TSSA) is a mucin-like molecule fromTrypanosoma cruzi, the etiological agent of Chagas disease, which displays amino acid polymorphisms in parasite isolates. TSSA expression is restricted to the surface of infective cell-derived trypomastigotes, where it functions as an adhesin and engages surface receptors on the host cell as a prerequisite for parasite internalization. Previous results have established TSSA-CL, the isoform encoded by the CL Brener clone, as an appealing candidate for use in serology-based diagnostics for Chagas disease. Here, we used a combination of peptide- and recombinant protein-based tools to map the antigenic structure of TSSA-CL at maximal resolution. Our results indicate the presence of different partially overlapping B-cell epitopes clustering in the central portion of TSSA-CL, which contains most of the polymorphisms found in parasite isolates. Based on these results, we assessed the serodiagnostic performance of a 21-amino-acid-long peptide that spans TSSA-CL major antigenic determinants, which was similar to the performance of the previously validated glutathioneS-transferase (GST)-TSSA-CL fusion molecule. Furthermore, the tools developed for the antigenic characterization of the TSSA antigen were also used to explore other potential diagnostic applications of the anti-TSSA humoral response in Chagasic patients. Overall, our present results provide additional insights into the antigenic structure of TSSA-CL and support this molecule as an excellent target for molecular intervention in Chagas disease.

scholarly journals Evaluation of a Prototype Flow Cytometry Test for Serodiagnosis of Canine Visceral Leishmaniasis

2013 ◽  
Vol 20 (12) ◽  
pp. 1792-1798 ◽  
Author(s):  
Henrique Gama Ker ◽  
Wendel Coura-Vital ◽  
Rodrigo Dian de Oliveira Aguiar-Soares ◽  
Bruno Mendes Roatt ◽  
Nádia das Dores Moreira ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Visceral Leishmaniasis ◽  
Cross Reactivity ◽  
Leishmania Braziliensis ◽  
Canine Visceral Leishmaniasis ◽  
Predictive Values ◽  
Content Type ◽  
Prototype Test ◽  
Serological Reactivity ◽  
Positive Results

ABSTRACTDiagnosing canine visceral leishmaniasis (CVL) is a critical challenge since conventional immunoserological tests still present some deficiencies. The current study evaluated a prototype flow cytometry serology test, using antigens and fluorescent antibodies that had been stored for 1 year at 4°C, on a broad range of serum samples. Noninfected control dogs andLeishmania infantum-infected dogs were tested, and the prototype test showed excellent performance in differentiating these groups with high sensitivity, specificity, positive and negative predictive values, and accuracy (100% in all analyses). When the CVL group was evaluated according to the dogs' clinical status, the prototype test showed outstanding accuracy in all groups with positive serology (asymptomatic II, oligosymptomatic, and symptomatic). However, in dogs which had positive results by PCR-restriction fragment length polymorphism (RFLP) but negative results by conventional serology (asymptomatic I), serological reactivity was not observed. Additionally, sera from 40 dogs immunized with different vaccines (Leishmune, Leish-Tec, or LBSap) did not present serological reactivity in the prototype test. Eighty-eight dogs infected with other pathogens (Trypanosoma cruzi,Leishmania braziliensis,Ehrlichia canis, andBabesia canis) were used to determine cross-reactivity and specificity, and the prototype test performed well, particularly in dogs infected withB. canisandE. canis(100% and 93.3% specificities, respectively). In conclusion, our data reinforce the potential of the prototype test for use as a commercial kit and highlight its outstanding performance even after storage for 1 year at 4°C. Moreover, the prototype test efficiently provided accurate CVL serodiagnosis with an absence of false-positive results in vaccinated dogs and minor cross-reactivity against other canine pathogens.

scholarly journals A Trypanosoma cruzi Small Surface Molecule Provides the First Immunological Evidence that Chagas' Disease Is Due to a Single Parasite Lineage

2002 ◽  
Vol 195 (4) ◽  
pp. 401-413 ◽  
Author(s):  
Javier M. Di Noia ◽  
Carlos A. Buscaglia ◽  
Claudia R. De Marchi ◽  
Igor C. Almeida ◽  
Alberto C.C. Frasch
Keyword(s):  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Cross Reactivity ◽  
Economic Problem ◽  
Future Research ◽  
Biological Traits ◽  
Small Surface ◽  
Genetic Lineages ◽  
Human Infections ◽  
Glycosylphosphatidyl Inositol

Chagas' disease is a major health and economic problem caused by the protozoan Trypanosoma cruzi. Multiple independently evolving clones define a complex parasite population that can be arranged into two broad genetic lineages termed T. cruzi I and II. These lineages have different evolutionary origin and display distinct ecological and biological traits. Here we describe a novel molecule termed TSSA for trypomastigote small surface antigen that provides the first immunological marker allowing discrimination between lineages. TSSA is a surface, glycosylphosphatidyl inositol (GPI)-anchored mucin-like protein, highly antigenic during the infection. TSSA sequences from different parasite isolates reveal a population dimorphism that perfectly matches with the two T. cruzi lineages. Interestingly, this dimorphism is restricted to the central region of the molecule, which comprises the immunodominant B cell epitopes. This sequence variability has a major impact on TSSA antigenicity, leading to no immunological cross-reactivity between both isoforms for antibodies present either in immunization or infection sera. Furthermore, the absolute seroprevalence for TSSA in confirmed Chagasic patients is restricted to T. cruzi II isoform, strongly suggesting that human infections are due to this particular subgroup. Even though association of T. cruzi II with Chagas' disease has been proposed based on molecular markers, this is the first immunological evidence supporting this hypothesis. The implications of these results for the future research on Chagas' disease could be envisaged.

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