Detection of Potentially Diagnostic Leishmanial Antigens by Western Blot Analysis of Sera from Patients with Kala-Azar or Multilesional Cutaneous Leishmaniasis

Bogdan, Christian; Stosiek, N; Fuchs, Harald; Röllinghoff, Martin; Solbach, Werner

doi:10.1093/infdis/162.6.1417

Cited by 23 publications

(22 citation statements)

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“…This method has used soluble extracts, 15,18,20,21,44 purified membrane proteins, 45 or recombinant proteins as antigens. [46][47][48] The immunoblot results obtained with soluble protein extracts of Leishmania species and strain 268T showed a complex reactivity pattern (Figures 1-4), in which most (93%) of the polypeptide fractions of parasites were recognized by at least one serum sample from the individuals tests.…”

Section: Discussionmentioning

confidence: 99%

“…55 A 42-kD antigen was identified by sera from patients with cutaneous and visceral leishmaniasis in an L. major extract. 21 The immune response induced by a recombinant protein from L (L.) amazonensis (33 kD [Larp33]) was also evaluated. Western blotting showed that Larp33 was a 40-kD protein expressed in L. (L.) amazonensis and L. (V.) brazilliensis.…”

Section: Discussionmentioning

confidence: 99%

“…63 It allows specific serodiagnosis of visceral and mucocutaneous leishmaniasis in patients living in nonendemic areas, and appears to be superior to the classic IFA. 21 The use of this test has been assayed in following chemotherapy and cure. 13 Our results showed that some proteins were specific for the serum samples from patients with Chagas' disease ( Table 2).…”

Section: Discussionmentioning

confidence: 99%

“…19 Crude extracts of cultured promastigote forms of Leishmania species are the current source of antigen for seroimmunologic assays in the clinical laboratory. [20][21][22] The parasites Trypanosoma cruzi and Leishmania species share antigenic determinants that cause significant cross-reactions in serologic tests, thus impairing accurate laboratory diagnosis of the specific infections. 23,24 Other trypanosomatid-derived antigens, which also share antigenic determinants with Leishmania, are easily produced and contain epitopes recognized by human antibodies.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis.

Gonçalves¹,

Reiche²,

Filho³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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Abstract. A Western blot method that uses antigens from culture promastigote forms of Leishmania (Viannia) braziliensis, L. (Leishmania) amazonensis, L. (Leishmania) tropica, and a trypanosomatid (strain 268T) isolated from naturally infected tomatoes was evaluated for laboratory diagnosis of American tegumentary leishmaniasis (ATL). Serum samples were obtained from 108 patients with ATL (group I), 23 chagasic patients (group II), 32 patients with other diseases (group III), and 78 healthy individuals (group IV). The overall analysis showed a sensitivity of 76.90%, 90.40%, 78.50%, and 87.90%, a specificity of 100%, 93.80%, 87.80%, and 77.10%, a positive predictive value of 100%, 94.00%, 89.50%, and 72.50%, a negative predictive value of 75.70%, 90.00%, 75.40%, and 90.20%, and a concordance coefficient kappa of 0.7358, 0.8400, 0.6491, and 0.6287 for L.(L.) tropica, and strain 268T antigens, respectively. The antigenic profile recognized by serum samples from patients with ATL and with Chagas' disease permits serologic distinction between these infections.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis.

Gonçalves¹,

Reiche²,

Filho³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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“…After three final washings, the cells were mounted with Molecular Probes ProLong Gold Antifade Mountant (Life Technologies), containing DAPI (4=,6-diamidino-2-phenylindole) to stain DNA, on coverslips and cured overnight. Mouse anti-HA Ab (diluted 1:500; clone 16B12; Covance), rat anti-LAMP-1 (diluted 1:600; clone 1D4B; DSHB), and polyclonal human antiLeishmania serum (diluted 1:1,000; derived from a patient with multilesional cutaneous leishmaniasis due to L. infantum infection [85]) were used as primary Abs. Goat anti-mouse IgG Ab coupled to Alexa 488 (diluted 1:400; Dianova), goat anti-mouse IgG Ab coupled to Alexa 594 (diluted 1:1,600; Dianova), donkey anti-rat IgG Ab coupled to Cy5 (diluted 1:400; Dianova), goat-anti-human IgG coupled to Alexa 488 (diluted 1:1,600; Jackson ImmunoResearch), and goat-anti-human IgG coupled to Alexa 647 (diluted 1:100; Jackson ImmunoResearch) were used as secondary Abs.…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

et al. 2017

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Similar to other intracellular pathogens, Leishmania parasites are known to evade the antimicrobial effector functions of host immune cells. To date, however, only a few virulence factors have been described for Leishmania major, one of the causative agents of cutaneous leishmaniasis. Here, we have characterized the expression and function of an L. major phosphatase, which we termed LmPRL-1. This enzyme shows a strong structural similarity to the human phosphatases of regenerating liver (PRL-1, -2, and -3) that regulate the proliferation, differentiation, and motility of cells. The biochemical characterization of the L. major phosphatase revealed that the enzyme is redox sensitive. When analyzing the subcellular localization of LmPRL-1 in promastigotes, amastigotes, and infected macrophages, we found that the phosphatase was predominantly expressed and secreted by promastigotes via the exosome route. Finally, we observed that ectopic expression of LmPRL-1 in L. major led to an increased number of parasites in macrophages. From these data, we conclude that the L. major phosphatase LmPRL-1 contributes to the intracellular survival of the parasites in macrophages.KEYWORDS Leishmania, exosome, macrophage, tyrosine phosphatase L eishmaniasis is an infectious disease prevalent worldwide that is caused by kinetoplastid protozoan parasites belonging to the genus Leishmania (1). In nature, this heteroxenous parasite is transmitted by the bites of sand flies. Following the inoculation of flagellated promastigotes into the dermis of the host, the parasites are rapidly endocytosed by phagocytic cells, in which they differentiate into amastigotes, multiply, and reach inner compartments and organs, such as draining lymph nodes, spleen, liver, and bone marrow. The life cycle of the parasite is completed after ingestion of amastigote-infected cells by sand flies during their blood meal. In the digestive tract of their vector, parasites transform back into extracellular promastigotes that develop into infectious metacyclics (2). Depending on the status of the host immune system and the Leishmania species, the infection of mammals leads either to mostly self-healing skin lesions (cutaneous leishmaniasis [CL]) or to a systemic disease, termed visceral leishmaniasis (VL) or kala-azar, that is lethal if untreated (3-5).To survive in their hosts, Leishmania parasites need to quickly adapt their growth, metabolism, and mechanisms of protection to their new environment. Interestingly, only a few genes are differentially expressed during this adaptive process, suggesting an important role for the regulation of protein translation (6). Leishmania speciesspecific gene diversity also participates in the adaptation of the parasite to its organ-

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Chronic Systemic Infection of Mice with Leishmania infantum Leads to Increased Bone Mass

Lai

Heinemann

Schleicher

et al. 2020

Journal of Bone and Mineral Research

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Vector‐borne infections of humans with the protozoan parasite Leishmania (L.) infantum can cause a systemic and potentially lethal disease termed visceral leishmaniasis. In the corresponding mouse model, an intravenous infection with L. infantum leads to the persistence of parasites in various organs, including bone marrow (BM). Considering the anatomical proximity between the BM and the cortical bone, we investigated whether a chronic infection with L. infantum affected bone homeostasis. Unexpectedly, chronic infection with L. infantum caused an increase in bone mass in mice. In vivo, an increased number of osteoblasts and osteocytes and a decreased maturation of osteoclasts characterized the phenotype. Confocal laser scanning fluorescence microscopy confirmed the infection of BM macrophages but also revealed the presence of parasites in osteoclasts. In vitro, mature osteoclasts took up L. infantum parasites. However, infection of osteoclast progenitors abolished their differentiation and function. In addition, secretory products of infected BM–derived macrophages inhibited the maturation of osteoclasts. Both in vitro and in vivo, infected macrophages and osteoclasts showed an enhanced expression of the anti‐osteoclastogenic chemokine CCL5 (RANTES). Neutralization of CCL5 prevented the inhibition of osteoclast generation seen in the presence of culture supernatants from L. infantum‐infected macrophages. Altogether, our study shows that chronic infection with Leishmania increases bone mass by inducing bone formation and impairing osteoclast differentiation and function. © 2022 American Society for Bone and Mineral Research (ASBMR).

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Detection of Potentially Diagnostic Leishmanial Antigens by Western Blot Analysis of Sera from Patients with Kala-Azar or Multilesional Cutaneous Leishmaniasis

Cited by 23 publications

References 0 publications

Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis.

Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis.

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

Chronic Systemic Infection of Mice with Leishmania infantum Leads to Increased Bone Mass

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