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Proteomics-Based Characterization of the Humoral Immune Response in Sporotrichosis: Toward Discovery of Potential Diagnostic and Vaccine Antigens.

Rodrigues AM, Fernandes GF, Araujo LM, Della Terra PP, dos Santos PO, Pereira SA, Schubach TM, Burger E, Lopes-Bezerra LM, de Camargo ZP - PLoS Negl Trop Dis (2015)

Bottom Line: Epidemiological surveillance has highlighted an overwhelming occurrence of the highly pathogenic fungus Sporothrix brasiliensis during feline outbreaks, leading to massive transmissions to humans.Antigen profiling included protein extracts from the closest known relatives S. brasiliensis and S. schenckii.We propose that 3-carboxymuconate cyclase has potential for the serological diagnosis of sporotrichosis and as target for the development of an effective multi-species vaccine against sporotrichosis in animals and humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Parasitology, Discipline of Cellular Biology, Federal University of São Paulo, São Paulo, Brazil.

ABSTRACT

Background: Sporothrix schenckii and associated species are agents of human and animal sporotrichosis that cause large sapronoses and zoonoses worldwide. Epidemiological surveillance has highlighted an overwhelming occurrence of the highly pathogenic fungus Sporothrix brasiliensis during feline outbreaks, leading to massive transmissions to humans. Early diagnosis of feline sporotrichosis by demonstrating the presence of a surrogate marker of infection can have a key role for selecting appropriate disease control measures and minimizing zoonotic transmission to humans.

Methodology: We explored the presence and diversity of serum antibodies (IgG) specific against Sporothrix antigens in cats with sporotrichosis and evaluated the utility of these antibodies for serodiagnosis. Antigen profiling included protein extracts from the closest known relatives S. brasiliensis and S. schenckii. Enzyme-linked immunosorbent assays and immunoblotting enabled us to characterize the major antigens of feline sporotrichosis from sera from cats with sporotrichosis (n = 49), healthy cats (n = 19), and cats with other diseases (n = 20).

Principal findings: Enzyme-linked immunosorbent assay-based quantitation of anti-Sporothrix IgG exhibited high sensitivity and specificity in cats with sporotrichosis (area under the curve, 1.0; 95% confidence interval, 0.94-1; P<0.0001) versus controls. The two sets of Sporothrix antigens were remarkably cross-reactive, supporting the hypothesis that antigenic epitopes may be conserved among closely related agents. One-dimensional immunoblotting indicated that 3-carboxymuconate cyclase (a 60-kDa protein in S. brasiliensis and a 70-kDa protein in S. schenckii) is the immunodominant antigen in feline sporotrichosis. Two-dimensional immunoblotting revealed six IgG-reactive isoforms of gp60 in the S. brasiliensis proteome, similar to the humoral response found in human sporotrichosis.

Conclusions: A convergent IgG-response in various hosts (mice, cats, and humans) has important implications for our understanding of the coevolution of Sporothrix and its warm-blooded hosts. We propose that 3-carboxymuconate cyclase has potential for the serological diagnosis of sporotrichosis and as target for the development of an effective multi-species vaccine against sporotrichosis in animals and humans.

No MeSH data available.


Related in: MedlinePlus

Representative immunoblot of S. brasiliensis (CBS 132990 and CBS 132021) and S. schenckii (CBS 132974 and CBS 132984) antigens using sera from cats with confirmed sporotrichosis.Despite the diversity of recognition patterns in different animals, the humoral response was consistently strong against the immunodominant antigen 3-carboxymuconate cyclase (gp60 in S. brasiliensis and gp70 in S. schenckii s. str.). Immunoblotting is as described in Methods.
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pntd.0004016.g003: Representative immunoblot of S. brasiliensis (CBS 132990 and CBS 132021) and S. schenckii (CBS 132974 and CBS 132984) antigens using sera from cats with confirmed sporotrichosis.Despite the diversity of recognition patterns in different animals, the humoral response was consistently strong against the immunodominant antigen 3-carboxymuconate cyclase (gp60 in S. brasiliensis and gp70 in S. schenckii s. str.). Immunoblotting is as described in Methods.

Mentions: As expected, antibodies from cats with sporotrichosis reacted with a wide variety of S. brasiliensis and S. schenckii proteins 20kDa to >160kDa in size (Fig 3). Cat-to-cat variation resulted in characteristic banding patterns for each animal (Fig 3); supporting the hypothesis that in a genetically diverse population, the antibody repertoire is expected to vary among individual cats. On the other hand, we detected minor or no differences in IgG-reacting banding patterns between antigen preparations (Fig 3), consistent with the close genetic distance between S. brasiliensis and S. schenckii [8, 9]. Despite this variation, all cats produced antibodies against a 60-kDa molecule in the S. brasiliensis proteome and a 70-kDa molecule in the S. schenckii proteome.


Proteomics-Based Characterization of the Humoral Immune Response in Sporotrichosis: Toward Discovery of Potential Diagnostic and Vaccine Antigens.

Rodrigues AM, Fernandes GF, Araujo LM, Della Terra PP, dos Santos PO, Pereira SA, Schubach TM, Burger E, Lopes-Bezerra LM, de Camargo ZP - PLoS Negl Trop Dis (2015)

Representative immunoblot of S. brasiliensis (CBS 132990 and CBS 132021) and S. schenckii (CBS 132974 and CBS 132984) antigens using sera from cats with confirmed sporotrichosis.Despite the diversity of recognition patterns in different animals, the humoral response was consistently strong against the immunodominant antigen 3-carboxymuconate cyclase (gp60 in S. brasiliensis and gp70 in S. schenckii s. str.). Immunoblotting is as described in Methods.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549111&req=5

pntd.0004016.g003: Representative immunoblot of S. brasiliensis (CBS 132990 and CBS 132021) and S. schenckii (CBS 132974 and CBS 132984) antigens using sera from cats with confirmed sporotrichosis.Despite the diversity of recognition patterns in different animals, the humoral response was consistently strong against the immunodominant antigen 3-carboxymuconate cyclase (gp60 in S. brasiliensis and gp70 in S. schenckii s. str.). Immunoblotting is as described in Methods.
Mentions: As expected, antibodies from cats with sporotrichosis reacted with a wide variety of S. brasiliensis and S. schenckii proteins 20kDa to >160kDa in size (Fig 3). Cat-to-cat variation resulted in characteristic banding patterns for each animal (Fig 3); supporting the hypothesis that in a genetically diverse population, the antibody repertoire is expected to vary among individual cats. On the other hand, we detected minor or no differences in IgG-reacting banding patterns between antigen preparations (Fig 3), consistent with the close genetic distance between S. brasiliensis and S. schenckii [8, 9]. Despite this variation, all cats produced antibodies against a 60-kDa molecule in the S. brasiliensis proteome and a 70-kDa molecule in the S. schenckii proteome.

Bottom Line: Epidemiological surveillance has highlighted an overwhelming occurrence of the highly pathogenic fungus Sporothrix brasiliensis during feline outbreaks, leading to massive transmissions to humans.Antigen profiling included protein extracts from the closest known relatives S. brasiliensis and S. schenckii.We propose that 3-carboxymuconate cyclase has potential for the serological diagnosis of sporotrichosis and as target for the development of an effective multi-species vaccine against sporotrichosis in animals and humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Immunology and Parasitology, Discipline of Cellular Biology, Federal University of São Paulo, São Paulo, Brazil.

ABSTRACT

Background: Sporothrix schenckii and associated species are agents of human and animal sporotrichosis that cause large sapronoses and zoonoses worldwide. Epidemiological surveillance has highlighted an overwhelming occurrence of the highly pathogenic fungus Sporothrix brasiliensis during feline outbreaks, leading to massive transmissions to humans. Early diagnosis of feline sporotrichosis by demonstrating the presence of a surrogate marker of infection can have a key role for selecting appropriate disease control measures and minimizing zoonotic transmission to humans.

Methodology: We explored the presence and diversity of serum antibodies (IgG) specific against Sporothrix antigens in cats with sporotrichosis and evaluated the utility of these antibodies for serodiagnosis. Antigen profiling included protein extracts from the closest known relatives S. brasiliensis and S. schenckii. Enzyme-linked immunosorbent assays and immunoblotting enabled us to characterize the major antigens of feline sporotrichosis from sera from cats with sporotrichosis (n = 49), healthy cats (n = 19), and cats with other diseases (n = 20).

Principal findings: Enzyme-linked immunosorbent assay-based quantitation of anti-Sporothrix IgG exhibited high sensitivity and specificity in cats with sporotrichosis (area under the curve, 1.0; 95% confidence interval, 0.94-1; P<0.0001) versus controls. The two sets of Sporothrix antigens were remarkably cross-reactive, supporting the hypothesis that antigenic epitopes may be conserved among closely related agents. One-dimensional immunoblotting indicated that 3-carboxymuconate cyclase (a 60-kDa protein in S. brasiliensis and a 70-kDa protein in S. schenckii) is the immunodominant antigen in feline sporotrichosis. Two-dimensional immunoblotting revealed six IgG-reactive isoforms of gp60 in the S. brasiliensis proteome, similar to the humoral response found in human sporotrichosis.

Conclusions: A convergent IgG-response in various hosts (mice, cats, and humans) has important implications for our understanding of the coevolution of Sporothrix and its warm-blooded hosts. We propose that 3-carboxymuconate cyclase has potential for the serological diagnosis of sporotrichosis and as target for the development of an effective multi-species vaccine against sporotrichosis in animals and humans.

No MeSH data available.


Related in: MedlinePlus