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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.The two sets of Sporothrix antigens were remarkably cross-reactive, supporting the hypothesis that antigenic epitopes may be conserved among closely related agents.

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

Frequency and diversity of serum-derived antibodies (IgG isotype) against S. brasiliensis and S. schenckii antigens in naturally infected cats.3-carboxymuconate cyclase (gp60 in the S. brasiliensis proteome (A); gp70 in the S. schenckii proteome (B)) is the immunodominant molecule in feline sporotrichosis; it was recognized by 100% of the cat sera tested here irrespective of disease form and severity. (C) Diversity of recognition of S. brasiliensis antigens (outer ring, CBS 132990; inner ring, CBS 132021). (D) Diversity of recognition of S. schenckii antigens (outer ring, CBS 132974; inner ring, CBS 132984). Charts are proportional. Molecular weights are colored as indicated. Further information about the frequency of antigen recognition appears in S2 Table.
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pntd.0004016.g004: Frequency and diversity of serum-derived antibodies (IgG isotype) against S. brasiliensis and S. schenckii antigens in naturally infected cats.3-carboxymuconate cyclase (gp60 in the S. brasiliensis proteome (A); gp70 in the S. schenckii proteome (B)) is the immunodominant molecule in feline sporotrichosis; it was recognized by 100% of the cat sera tested here irrespective of disease form and severity. (C) Diversity of recognition of S. brasiliensis antigens (outer ring, CBS 132990; inner ring, CBS 132021). (D) Diversity of recognition of S. schenckii antigens (outer ring, CBS 132974; inner ring, CBS 132984). Charts are proportional. Molecular weights are colored as indicated. Further information about the frequency of antigen recognition appears in S2 Table.

Mentions: The major antigenic S. brasiliensis molecules (CBS 132990 and CBS 132021) recognized by feline IgG consisted of the following sizes: 60 kDa (100% and 100%, respectively), 90 kDa (92% and 92%, respectively), 100 kDa (86% and 86%, respectively), 38 kDa (60% and 56%, respectively), 40 kDa (56% and 58%, respectively), 45 kDa (44% and 42%, respectively), 30 kDa (36% and 26%, respectively), 52 kDa (30% and 32%, respectively), and 110 kDa (28% and 30%, respectively) (Fig 4A and 4C). Minor molecules recognized by feline IgG had sizes of 80 kDa, 25 kDa, 28 kDa, 120 kDa, 160 kDa, 35 kDa, 20 kDa, 55 kDa, 85 kDa, and 23 kDa (Fig 4A and 4C). The major antigenic S. schenckii molecules (CBS 132974 and CBS 132984) recognized by feline IgG had sizes of: 70 kDa (100% and 100%, respectively), 90 kDa (86% and 88%, respectively), 100 kDa (76% and 82%, respectively), 38 kDa (74% and 62%, respectively), 40 kDa (64% and 56%, respectively), 52 kDa (58% and 56%, respectively), 30 kDa (50% and 34%, respectively), 55 kDa (48% and 48%, respectively), and 45 kDa (40% and 30%, respectively) (Fig 4B and 4D). The minor S. schenckii molecules recognized by feline IgG had sizes of 25 kDa, 80 kDa, 28 kDa, 110 kDa, 120 kDa, 23 kDa, 35 kDa, 160 kDa, 85 kDa, and 20 kDa (Fig 4B and 4D). Sera from uninfected cats did not react with S. brasiliensis or S. schenckii antigens. Sera from cats with other infections were also non-reactive in the immunoblot assay. The frequencies at which Sporothrix molecules were recognized in the antigen preparations are presented in S2 Table. There was no association between the number of bands recognized by each serum and the distribution and number of skin lesions on cats with sporotrichosis.


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)

Frequency and diversity of serum-derived antibodies (IgG isotype) against S. brasiliensis and S. schenckii antigens in naturally infected cats.3-carboxymuconate cyclase (gp60 in the S. brasiliensis proteome (A); gp70 in the S. schenckii proteome (B)) is the immunodominant molecule in feline sporotrichosis; it was recognized by 100% of the cat sera tested here irrespective of disease form and severity. (C) Diversity of recognition of S. brasiliensis antigens (outer ring, CBS 132990; inner ring, CBS 132021). (D) Diversity of recognition of S. schenckii antigens (outer ring, CBS 132974; inner ring, CBS 132984). Charts are proportional. Molecular weights are colored as indicated. Further information about the frequency of antigen recognition appears in S2 Table.
© Copyright Policy
Related In: Results  -  Collection

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

pntd.0004016.g004: Frequency and diversity of serum-derived antibodies (IgG isotype) against S. brasiliensis and S. schenckii antigens in naturally infected cats.3-carboxymuconate cyclase (gp60 in the S. brasiliensis proteome (A); gp70 in the S. schenckii proteome (B)) is the immunodominant molecule in feline sporotrichosis; it was recognized by 100% of the cat sera tested here irrespective of disease form and severity. (C) Diversity of recognition of S. brasiliensis antigens (outer ring, CBS 132990; inner ring, CBS 132021). (D) Diversity of recognition of S. schenckii antigens (outer ring, CBS 132974; inner ring, CBS 132984). Charts are proportional. Molecular weights are colored as indicated. Further information about the frequency of antigen recognition appears in S2 Table.
Mentions: The major antigenic S. brasiliensis molecules (CBS 132990 and CBS 132021) recognized by feline IgG consisted of the following sizes: 60 kDa (100% and 100%, respectively), 90 kDa (92% and 92%, respectively), 100 kDa (86% and 86%, respectively), 38 kDa (60% and 56%, respectively), 40 kDa (56% and 58%, respectively), 45 kDa (44% and 42%, respectively), 30 kDa (36% and 26%, respectively), 52 kDa (30% and 32%, respectively), and 110 kDa (28% and 30%, respectively) (Fig 4A and 4C). Minor molecules recognized by feline IgG had sizes of 80 kDa, 25 kDa, 28 kDa, 120 kDa, 160 kDa, 35 kDa, 20 kDa, 55 kDa, 85 kDa, and 23 kDa (Fig 4A and 4C). The major antigenic S. schenckii molecules (CBS 132974 and CBS 132984) recognized by feline IgG had sizes of: 70 kDa (100% and 100%, respectively), 90 kDa (86% and 88%, respectively), 100 kDa (76% and 82%, respectively), 38 kDa (74% and 62%, respectively), 40 kDa (64% and 56%, respectively), 52 kDa (58% and 56%, respectively), 30 kDa (50% and 34%, respectively), 55 kDa (48% and 48%, respectively), and 45 kDa (40% and 30%, respectively) (Fig 4B and 4D). The minor S. schenckii molecules recognized by feline IgG had sizes of 25 kDa, 80 kDa, 28 kDa, 110 kDa, 120 kDa, 23 kDa, 35 kDa, 160 kDa, 85 kDa, and 20 kDa (Fig 4B and 4D). Sera from uninfected cats did not react with S. brasiliensis or S. schenckii antigens. Sera from cats with other infections were also non-reactive in the immunoblot assay. The frequencies at which Sporothrix molecules were recognized in the antigen preparations are presented in S2 Table. There was no association between the number of bands recognized by each serum and the distribution and number of skin lesions on cats with sporotrichosis.

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.The two sets of Sporothrix antigens were remarkably cross-reactive, supporting the hypothesis that antigenic epitopes may be conserved among closely related agents.

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