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.

The use of heterologous radioimmunoassays for the measurement of follicle-stimulating hormone and luteinizing hormone concentrations in horse and donkey serum

1983 ◽  
Vol 99 (2) ◽  
pp. 199-209 ◽  
Author(s):  
Valerie Urwin
Keyword(s):  
Luteinizing Hormone ◽  
Follicle Stimulating Hormone ◽  
Cross Reactivity ◽  
Chorionic Gonadotrophin ◽  
Cross Reaction ◽  
The Other ◽  
Corpora Lutea ◽  
Serum Concentrations ◽  
Study Support ◽  
Baseline Levels

Heterologous double-antibody radioimmunoassays were developed for the measurement of FSH and LH concentrations in the serum of both horses and donkeys. The FSH assay employed a rabbit anti-ovine FSH serum which showed a complete lack of cross-reaction with equine chorionic gonadotrophin (eCG) and negligible cross-reaction with equine LH. The LH assay utilized an antiserum raised against highly purified eCG. This similarly showed negligible cross-reaction with equine FSH but its high cross-reactivity with eCG prevented the measurement of equine LH concentrations in serum when eCG was also present. In both assays serial dilutions of horse and donkey serum were parallel to the standard. The assays were used to monitor changes in serum concentrations of FSH and LH during the first 100 days of pregnancy in pony mares and jenny donkeys. In both species during pregnancy LH levels reached a peak 1–2 days after ovulation. They then decreased rapidly to baseline levels where they remained until days 35–40 when the commencement of eCG production prevented their further measurement. Serum FSH concentrations, on the other hand, continued to fluctuate markedly throughout the first 100 days of pregnancy in both the ponies and donkeys. Pronounced surges in FSH levels occurred at regular intervals in some animals but the pattern of release was quite irregular in the others. The results of this study support the concept that it is continued pituitary FSH release, not eCG secretion, which is responsible for stimulating the secondary follicles which develop during early equine pregnancy. However, it appears likely that it is the LH-like activity of eCG which causes the subsequent ovulation and/or luteinization of these secondary follicles to produce accessory corpora lutea.

scholarly journals Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting)

1990 ◽  
Vol 32 (5) ◽  
pp. 379-383 ◽  
Author(s):  
Anna Maria Simonsen Stolf ◽  
Eufrosina Setsu Umezawa ◽  
Bianca Zingales
Keyword(s):  
Trypanosoma Cruzi ◽  
Western Blot ◽  
Chagas Disease ◽  
Western Blotting ◽  
Specific Antibodies ◽  
Internal Parasite ◽  
Sds Page ◽  
Blotting Technique ◽  
Simultaneous Identification ◽  
Acute Chagas Disease

A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained.

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 Analysis of the specificity of human antibodies to antigens of Leishmania braziliensis braziliensis

1989 ◽  
Vol 31 (4) ◽  
pp. 228-234 ◽  
Author(s):  
Aoi Masuda ◽  
Sueli Fátima do Nascimento ◽  
Carmem Silvia Guerra ◽  
Gláucia da Silva Paranhos ◽  
Antonio Walter Ferreira
Keyword(s):  
Visceral Leishmaniasis ◽  
Cutaneous Leishmaniasis ◽  
Chagas Disease ◽  
The Other ◽  
Leishmania Braziliensis ◽  
Human Antibodies ◽  
American Cutaneous Leishmaniasis

The antigenicity of promastigotes of Leishmania braziliensis braziliensis (L. b.braziliensis) treated with 1% sodium desoxycholate in 10 mM Tris-Hcl pH 8.2 was analysed by immunoblot using as probes sera from American cutaneous leishmaniasis (ACL), American visceral leishmaniasis (AVL), schistosomiasis, malaria and Chagas' disease. The ACL sera reacted constantly with a 60 kD band. No reactivity to this protein was observed with sera from the other diseases above mentioned indicating that the 60 kD protein may be used in serodiagnosis for ACL.

scholarly journals Cross-Reactivity Using Chimeric Trypanosoma cruzi Antigens: Diagnostic Performance in Settings Where Chagas Disease and American Cutaneous or Visceral Leishmaniasis Are Coendemic

2019 ◽  
Vol 57 (8) ◽  
Author(s):  
Ramona Tavares Daltro ◽  
Leonardo Maia Leony ◽  
Natália Erdens Maron Freitas ◽  
Ângelo Antônio Oliveira Silva ◽  
Emily Ferreira Santos ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Chagas Disease ◽  
Diagnostic Performance ◽  
Latent Class ◽  
Cross Reactivity ◽  
Chimeric Proteins ◽  
Serum Samples ◽  
Perfect Agreement ◽  
Content Type ◽  
Chronic Chagas Disease

ABSTRACT Chimeric T. cruzi antigens have been proposed as a diagnostic tool for chronic Chagas disease (CD) in both settings where Chagas disease is endemic and those where it is not endemic. Antibody response varies in accordance to each T. cruzi strain, presenting challenges to the use of antigens lacking demonstrated cross-reactivity with Leishmania spp. Our group expressed four chimeric proteins (IBMP-8.1, IBMP-8.2, IBMP-8.3, and IBMP-8.4) and previously assessed their diagnostic performance to determine cross-reactivity with Leishmania spp. Here, we validated our findings using serum samples from different Brazilian geographic areas reporting endemic Chagas disease, endemic visceral or American cutaneous leishmaniasis (ACL), or both. Overall, 829 serum samples were evaluated using commercial and IBMP enzyme-linked immunosorbent assays. Due to the absence of a reference assay to diagnosis CD, latent class analysis (LCA) was performed through the use of a statistical model. The incidence of cross-reactivity for ACL-positive samples varied from 0.35% (IBMP-8.3) to 0.70% (IBMP-8.1 and IBMP-8.2). Regarding visceral leishmaniasis (VL)-positive samples, the IBMP-8.2 and IBMP-8.3 antigens cross-reacted with six (3.49%) and with only one sample (0.58%), respectively. No cross-reactivity with either ACL or VL was observed for the IBMP-8.4 antigen. Similarly, no cross-reactions were found when VL-positive samples were assayed with IBMP-8.1. The agreement among the results obtained using IBMP antigens ranged from 97.3% for IBMP-8.2 and 99% for IBMP-8.1 and IBMP-8.3 to 100% for IBMP-8.4, demonstrating almost perfect agreement with LCA. Accordingly, in light of the negligible cross-reactivity with both ACL and VL, we suggest the use of IBMP antigens in regions where T. cruzi and Leishmania spp. are coendemic.

scholarly journals Leishmania major-Like Antigen for Specific and Sensitive Serodiagnosis of Human and Canine Visceral Leishmaniasis

2002 ◽  
Vol 9 (6) ◽  
pp. 1361-1366 ◽  
Author(s):  
Rosângela Barbosa-de-Deus ◽  
Marcos Luíz dos Mares-Guia ◽  
Adriane Zacarias Nunes ◽  
Kátia Morais Costa ◽  
Roberto Gonçalves Junqueira ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Rheumatoid Factor ◽  
Chagas Disease ◽  
Leishmania Major ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitological Diagnosis ◽  
Human Sera ◽  
Indirect Immunofluorescent

ABSTRACT An antigen (LMS) prepared from Leishmania major-like promastigotes was used in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of human and dog visceral leishmaniasis. The results were compared with those from the indirect immunofluorescent antibody test (IFAT). A total of 1,822 canine sera were tested, including sera from dogs with visceral leishmaniasis, transmissible venereal tumors, ehrlichiosis, rickettsiosis, or Chagas' disease and sera from healthy dogs. The antigen was also tested with 227 samples of human sera, including sera from patients with visceral, cutaneous, or diffuse cutaneous leishmaniasis and from noninfected individuals, as well as sera from patients with Chagas' disease, toxoplasmosis, rickettsiosis, hepatitis B, schistosomiasis, ascaridiasis, malaria, rheumatoid factor, leprosy and rheumatoid factor, tuberculosis, or leprosy. All dogs and all human patients had a clinical and/or serological and/or parasitological diagnosis. For detecting antibodies in sera from dogs with leishmaniasis, the antigen showed a sensitivity of 98%, specificity of 95%, and concordance of 93% and when used for detecting antibodies in human sera presented a sensitivity of 92%, specificity of 100%, and concordance of 92%. Comparison between ELISA and IFAT demonstrated that ELISA using the LMS antigen yielded more reliable results than IFAT. The LMS antigen displayed no cross-reactivity with sera from patients or dogs that had any of the other diseases tested.

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.

Screening diagnostic candidates from Leishmania infantum proteins for human visceral leishmaniasis using an immunoproteomics approach

Parasitology ◽  
2019 ◽  
Vol 146 (11) ◽  
pp. 1467-1476 ◽  
Author(s):  
Daniela P. Lage ◽  
Fernanda Ludolf ◽  
Patrícia C. Silveira ◽  
Amanda S. Machado ◽  
Fernanda F. Ramos ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Infantum ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Diagnostic Tools ◽  
Endemic Region ◽  
Endonuclease Iii ◽  
Human Visceral Leishmaniasis ◽  
Variable Sensitivity ◽  
Epidemiological Evaluation

AbstractThere is no suitable vaccine against human visceral leishmaniasis (VL) and available drugs are toxic and/or present high cost. In this context, diagnostic tools should be improved for clinical management and epidemiological evaluation of disease. However, the variable sensitivity and/or specificity of the used antigens are limitations, showing the necessity to identify new molecules to be tested in a more sensitive and specific serology. In the present study, an immunoproteomics approach was performed in Leishmania infantum promastigotes and amastigotes employing sera samples from VL patients. Aiming to avoid undesired cross-reactivity in the serological assays, sera from Chagas disease patients and healthy subjects living in the endemic region of disease were also used in immunoblottings. The most reactive spots for VL samples were selected, and 29 and 21 proteins were identified in the promastigote and amastigote extracts, respectively. Two of them, endonuclease III and GTP-binding protein, were cloned, expressed, purified and tested in ELISA experiments against a large serological panel, and results showed high sensitivity and specificity values for the diagnosis of disease. In conclusion, the identified proteins could be considered in future studies as candidate antigens for the serodiagnosis of human VL.

scholarly journals Hemolymphatic components in vectors of Trypanosoma cruzi: study in several species of the subfamily Triatominae (Hemiptera: Reduviidae)

1993 ◽  
Vol 35 (2) ◽  
pp. 123-128 ◽  
Author(s):  
Lilián Etelvina Canavoso ◽  
Edilberto René Rubiolo
Keyword(s):  
Chemical Composition ◽  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Protein Composition ◽  
The Other ◽  
Adult Males ◽  
American Trypanosomiasis ◽  
Number Of Species ◽  
Medical Importance ◽  
Lipoprotein Profiles

The members of the subfamily Triatominae (Hemiptera : Reduviidae) comprise a great number of species of medical importance in the transmission of the T. cruzi (American trypanosomiasis). The aim of this study was to contribute to the knowledge about the chemical composition in proteins, lipids, lipoproteins, and carbohydrates of vectors of Chagas' disease corresponding to twelve members of the subfamily Triatominae. This study was carried out in ninphs of the fifth instar and adult males of the species: T. delpontei, T. dimidiata, T. guasayana, T. infestans, T. mazzotti, T. pallidipennis, T. patagonica, T. platensis, T. rubrovaria, T. sordida of the Triatoma genus, and D. maximus and P. megistus of the Dipatalogaster and Panstrongylus genera respectively. The results show on one hand, qualitative differences in the protein composition, and on the other hand, similarity in the lipoprotein profiles. Lipids, proteins, and carbohydrates did not show significant differences between species or/and stages.

Sign in / Sign up

Export Citation Format

Share Document