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1.
Chinese Journal of Epidemiology ; (12): 629-635, 2023.
Article in Chinese | WPRIM | ID: wpr-985538

ABSTRACT

Objective: The docking and superantigen activity sites of staphylococcal enterotoxin-like W (SElW) and T cell receptor (TCR) were predicted, and its SElW was cloned, expressed and purified. Methods: AlphaFold was used to predict the 3D structure of SElW protein monomers, and the protein models were evaluated with the help of the SAVES online server from ERRAT, Ramachandran plot, and Verify_3D. The ZDOCK server simulates the docking conformation of SElW and TCR, and the amino acid sequences of SElW and other serotype enterotoxins were aligned. The primers were designed to amplify selw, and the fragment was recombined into the pMD18-T vector and sequenced. Then recombinant plasmid pMD18-T was digested with BamHⅠand Hind Ⅲ. The target fragment was recombined into the expression plasmid pET-28a(+). After identification of the recombinant plasmid, the protein expression was induced by isopropyl-beta-D- thiogalactopyranoside. The SElW expressed in the supernatant was purified by affinity chromatography and quantified by the BCA method. Results: The predicted three-dimensional structure showed that the SElW protein was composed of two domains, the amino-terminal and the carboxy-terminal. The amino-terminal domain was composed of 3 α-helices and 6 β-sheets, and the carboxy-terminal domain included 2 α-helices and 7 antiparallel β-sheets composition. The overall quality factor score of the SElW protein model was 98.08, with 93.24% of the amino acids having a Verify_3D score ≥0.2 and no amino acids located in disallowed regions. The docking conformation with the highest score (1 521.328) was selected as the analysis object, and the 19 hydrogen bonds between the corresponding amino acid residues of SElW and TCR were analyzed by PyMOL. Combined with sequence alignment and the published data, this study predicted and found five important superantigen active sites, namely Y18, N19, W55, C88, and C98. The highly purified soluble recombinant protein SElW was obtained with cloning, expression, and protein purification. Conclusions: The study found five superantigen active sites in SElW protein that need special attention and successfully constructed and expressed the SElW protein, which laid the foundation for further exploration of the immune recognition mechanism of SElW.


Subject(s)
Humans , Enterotoxins/genetics , Superantigens/genetics , Catalytic Domain , Selenoprotein W/metabolism , Receptors, Antigen, T-Cell
2.
Article in English | WPRIM | ID: wpr-762161

ABSTRACT

Staphylococcus aureus commonly colonizes the skin of atopic dermatitis (AD) patients and contributes to the development and exacerbation of AD. Multiple factors are associated with colonization of AD skin by S. aureus, including the strength of S. aureus-corneocyte adhesion, deficiency of antimicrobial peptides, decreased levels of filaggrin and filaggrin degradation products, overexpressed Th2/Th17 cytokines, microbial dysbiosis and altered lipid profiles. S. aureus colonization on AD skin causes skin barrier dysfunction through virulence factors such as superantigens (toxins), enzymes and other proteins. Furthermore, colonization of AD skin by S. aureus exacerbates AD and may contribute to microbial dysbiosis, allergen sensitization, Th2/Th17 polarization, development of atopic march and food allergy in AD patients. Skin colonization of S. aureus, particularly methicillin-resistant S. aureus (MRSA), is one of the major challenges commonly encountered in the management of AD. Bleach bath, and topical or systemic antibiotics could be used to control S. aureus infection on AD skin. However, careful use of antibiotics is required to control the occurence of MRSA. Recently, various strategies, including microbiome transplant, monoclonal antibodies against virulent toxins, vaccines and recombinant phage endolysin, have been studied to control S. aureus infection on AD skin. Further advances in our understanding of S. aureus could provide us with ways to manage S. aureus colonization more effectively in AD patients.


Subject(s)
Humans , Anti-Bacterial Agents , Antibodies, Monoclonal , Bacteriophages , Baths , Colon , Cytokines , Dermatitis, Atopic , Dysbiosis , Food Hypersensitivity , Methicillin Resistance , Methicillin-Resistant Staphylococcus aureus , Microbiota , Peptides , Skin , Staphylococcus aureus , Staphylococcus , Superantigens , Vaccines , Virulence Factors
3.
Biomédica (Bogotá) ; Biomédica (Bogotá);38(1): 96-104, ene.-mar. 2018. tab
Article in Spanish | LILACS | ID: biblio-888552

ABSTRACT

Resumen Introducción. Staphylococcus aureus coloniza mucosas y piel, y causa graves infecciones en el hombre y los animales. Es importante establecer el estatus de portadoras de cepas enterotoxigénicas de este microorganismo en manipuladoras de alimentos, con el fin de prevenir intoxicaciones alimentarias. Objetivo. Establecer las correlaciones entre los genes de enterotoxinas clásicas, el gen tsst-1, la producción de toxinas en cultivo y la resistencia antimicrobiana en aislamientos de S. aureus provenientes de manipuladoras de alimentos que cuidan niños en sus comunidades. Materiales y métodos. Se cultivaron muestras de las fosas nasales y las yemas de los dedos de las manos, y se identificó S. aureus empleando las pruebas de rutina y métodos automatizados. La extracción de ADN se hizo mediante el método de bromuro de cetil-trimetil-amonio (Cetyl-Trimethyl- Ammonium Bromide, CTAB) modificado. Para la detección de superantígenos se emplearon pruebas de reacción en cadena de la polimerasa (PCR) simple y múltiple, y para la de toxinas, estuches comerciales. Resultados. Se encontró que el 22,0 % de los aislamientos correspondía a portadoras de S. aureus: 17,0 % en los aislamientos de fosas nasales; 5,0 % en los de las manos y 6,7 % simultáneamente en los dos sitios. La prevalencia de superantígenos fue de 73,7 %. El genotipo más frecuente fue el sea-tsst-1, con 10,0 %. La resistencia a un solo antibiótico fue de 74,7 % y, a cuatro antibióticos, de 3,2 %; de los aislamientos, el 93,7 % correspondía a cepas productoras de betalactamasas. La detección de genes clásicos y de tsst-1 mediante PCR fue de 48,4 % y la de toxinas en el sobrenadante, de 42,1 %, con una correlación de 95,7 %. Las mayores correlaciones se establecieron entre las toxinas TSST-1 (22/22) y SEA (17/18). La correlación del gen tsst-1 con la proteína y la resistencia fue de 100 %. Todos los aislamientos con el genotipo sea-tsst-1 t fueron resistentes y productores de las toxinas. Conclusión. La tasa de aislamientos de S. aureus toxigénicos y resistentes obtenidos de mujeres que cuidan y preparan alimentos para niños fue de más de 70 %, lo que demostró su gran virulencia y la consecuente necesidad de aplicar estrictamente las normas higiénicas y sanitarias vigentes para evitar el riesgo de intoxicación alimentaria.


Abstract Introduction: Staphylococcus aureus colonizes mucous membranes and skin causing severe infections in humans and animals. It is important to determine carrier status of enterotoxigenic strains of this microorganism in food handlers to prevent food poisoning. Objective: To establish the correlations among classic enterotoxigenic genes, tsst-1 gene, the production of toxins in cultures and antimicrobial resistance in S. aureus isolates from women who handle the food, feed and take care of children in their communities. Materials and methods: Nasal swab and finger samples were cultured and S. aureus was identified using routine methods and automated systems. DNA extraction was done by the CTAB modified method, and superantigen detection by simple and multiplex PCR, while toxins were detected using commercial kits. Results: We found that 22.0% of subjects were S. aureus carriers: 17.0% corresponded to nose samples, 5.0% to hands and 6.7% to both nose and hands. The prevalence of superantigens was 73.7%. The most frequent genotype was sea-tsst-1 with 10%. Resistance to one antibiotic was 74.7%, and to four antibiotics, 3.2%; 93.7% of the isolates were betalactamase-positive. Classical genes and tsst-1 gene were detected by PCR in 48.4% of samples and toxins in supernatant were detected in 42.1% of them with 95.7% of correlation.The highest correlations were established for TSST-1 and SEA with 100% and 94.4%, respectively. The correlation of tsst-1 gene with toxin production and resistance was 100%. All isolates with genotype sea-tsst-1 were toxin-positive and resistant. Conclusion: The rate of toxigenic and resistant S. aureus isolates from women in charge of feeding and taking care of children was higher than 70%, which demonstrates its high virulence. This requires the strict application of hygienic and sanitary regulations in order to avoid the risk of food poisoning.


Subject(s)
Adult , Child , Female , Humans , Staphylococcal Infections/microbiology , Staphylococcus aureus/isolation & purification , Carrier State/microbiology , Child Care , Superantigens/analysis , Drug Resistance, Multiple, Bacterial , Enterotoxins/immunology , Antigens, Bacterial/analysis , Staphylococcal Infections/transmission , Staphylococcal Infections/epidemiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics , Staphylococcus aureus/immunology , Carrier State/epidemiology , Prevalence , Superantigens/genetics , Fingers/microbiology , Food Handling , Genes, Bacterial , Genotype , Nasal Cavity/microbiology , Antigens, Bacterial/genetics
4.
Chinese Journal of Epidemiology ; (12): 1375-1380, 2018.
Article in Chinese | WPRIM | ID: wpr-738155

ABSTRACT

Objective: To analyze the characteristics of super-antigen (SAg) of group A Streptococcus pyogenes (GAS), isolated from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017. Methods: Throat swab specimens from patients with scarlet fever or pharyngeal infections were collected and tested for GAS. Eleven currently known SAg genes including SpeA, speC, speG, speH, speI, speJ, speK, speL, speM, smeZ and ssa were tested by real-time PCR while M protein genes (emm genes) were amplified and sequenced by PCR. Results: A total of 377 GAS were isolated from 6 801 throat swab specimens, with the positive rate as 5.5%. There were obvious changes noticed among speC, speG, speH and speK in three years. A total of 45 SAg genes profiles were observed, according to the SAgs inclusion. There were significant differences appeared in the frequencies among two of the highest SAg genes profiles between emm1 and emm12 strains (χ(2)=38.196, P<0.001; χ(2)=72.310, P<0.001). There also appeared significant differences in the frequencies of speA, speH, speI and speJ between emm1 and emm12 strains (χ(2)=146.154, P<0.001; χ(2)=52.31, P<0.001; χ(2)=58.43, P<0.001; χ(2)=144.70, P<0.001). Conclusions: Obvious changes were noticed among SAg genes including speC, speG, speH and speK from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017. SAg genes including speA, speH, speI and speJ appeared to be associated with the emm 1 and emm 12 strains. More kinds of SAg genes profiles were isolated form GAS but with no significant differences seen in the main SAg genes profiles, during the epidemic period.


Subject(s)
Female , Humans , Pregnancy , Antigens, Bacterial/genetics , Bacterial Outer Membrane Proteins , Bacterial Proteins , Beijing/epidemiology , China/epidemiology , Exotoxins , Membrane Proteins , Pharyngitis/microbiology , Pharynx/microbiology , Pregnancy Complications, Infectious/microbiology , Real-Time Polymerase Chain Reaction , Scarlet Fever/microbiology , Streptococcal Infections , Streptococcus pyogenes/isolation & purification , Superantigens/genetics
5.
Article in English | WPRIM | ID: wpr-713331

ABSTRACT

OBJECTIVES: Pathophysiology of chronic rhinosinusitis (CRS) is very complex and has not yet been clearly understood. To date, various factors have been researched to have relations with the pathogenesis of CRS, such as superantigens and biofilms. Recently, we found an unusual pathological finding in patients with CRS, and we called this new entity as bacteria ball (or bioball). In this study, we analyze the clinical characteristics of bacteria ball occurred in CRS. METHODS: This study enrolled consecutive 247 patients with CRS who underwent functional endoscopic sinus surgery from January 2015 to August 2016. The diagnosis of bacterial ball was made when negative in Gomori-methenamine-silver stain and positive in Gram stain. Histologically, bacterial ball was defined as acellular mucous materials with bacterial colonies and inflammatory cell infiltrates. We compared clinical data and computed tomography (CT) findings between fungal and bacterial balls. RESULTS: Six cases (2.4%) of CRS were confirmed histologically as bacterial ball. Most of them were found in the maxillary sinus of CRS without nasal polyposis (66.7%). Bacterial ball was green or brown colored materials similar to fungal ball which was harder and tightly adherent to the antral mucosa. Compared to fungal ball, patients with bacterial ball showed significantly less peripheral eosinophils (P=0.011) and calcification in CT scans (P=0.003). CONCLUSION: Bacterial ball is unusual findings occurred in patient with CRS which is different from fungal ball and biofilm. For diagnosis of bacterial ball, Gram stain is essentially required to identify bacterial colonies. Bacterial ball might appear to be evidence of a new strategy for living in the paranasal sinuses.


Subject(s)
Humans , Bacteria , Biofilms , Diagnosis , Eosinophils , Fungi , Maxillary Sinus , Mucous Membrane , Paranasal Sinuses , Sinusitis , Superantigens , Tomography, X-Ray Computed
6.
Pesqui. vet. bras ; Pesqui. vet. bras;37(7): 691-696, jul. 2017. tab, ilus, mapas
Article in Portuguese | LILACS, VETINDEX | ID: biblio-895474

ABSTRACT

A capacidade de produção de toxinas pelo Staphylococcus aureus no leite e produtos derivados está relacionado com surtos de intoxicação alimentar. Objetivou-se nesta pesquisa, estudar a ocorrência de genes que codificam para enterotoxinas estafilocócicas (sea, seb, sed, seg, seh e sei) e toxinas α e ß hemolítica (hla e hlb) em S. aureus isolados de 53 amostras de leite de tanques expansão comunitários no Estado de Alagoas, Brasil. Foram identificados 27 isolados (50,94%) como S. aureus pela amplificação do gene nuc. 13/27 isolados (48,1%) foram positivos para pelo menos um gene das enterotoxinas estudadas, sendo as frequências dos genes sea 33,3%, seh 18,5%, sei 11,1% e sed 7,4%; não entanto não foram identificados os genes seb e seg nestas bactérias. Para as toxinas hemolíticas, 51,9% dos isolados portavam ambos genes (hla e hlb), sendo a frequência para o gene hla de 81,5% e para o gene hlb de 51,9%. A frequência de genes das toxinas avaliadas é alta o que constitui um risco potencial para a saúde pública em especial, as enterotoxinas por serem termoestáveis e estarem asssociados com surtos de intoxicação alimentar.(AU)


The capacity of toxin production by Staphylococcus aureus in milk and dairy products is associated with food poisoning outbreaks. The objective of this research was to study the frequency of genes encoding staphylococcal enterotoxin (sea, seb, sed, seg, seh and sei) and α and ß hemolytic toxins (hla and hlb) in S. aureus isolates from 53 milk samples from community tanks in the State of Alagoas, Brazil. Twenty-seven isolates (50.94%) were identified as S. aureus by nuc gene amplification; 13/27 isolates (48.1%) were positive for at least one gene of the studied enterotoxins and the frequency of genes sea was 33.3%, seh 18.5%, sei 11.1% and sed 7.4%; the seb and sec genes have not been identified in the bacteria. For the hemolytic toxins, 51.9% of isolates harbored both genes (hla and hlb), the frequency of hla gene was 81.5% and 51.9% for the hlb gene. The evaluated toxin-encoding gene frequency is high and constitutes a potential risk for public health, especially staphylococcal enterotoxin genes; because they are heat-stable enterotoxins and have been associated with food poisoning.(AU)


Subject(s)
Staphylococcus aureus/genetics , Superantigens/genetics , Milk/microbiology , Enterotoxins , Polymerase Chain Reaction/veterinary
8.
Prensa méd. argent ; Prensa méd. argent;103(6): 321-330, 20170000.
Article in Spanish | LILACS, BINACIS | ID: biblio-1377934

ABSTRACT

Los superantígenos (Sags) son proteínas de origen bacteriano o viral capaces de estimular y causar la muerte de un alto numero de linfocitos T o B. normales. Este hecho se debe a que ­ a diferencia de los antígenos convencionales que son reconocidos por todas las regiones variables del receptor de las células T (TCR) o B (BCR) - sólo interactúan con regiones constantes ubicadas dentro de las regiones variables de estos receptores. Los Sags B se unen a sitios constantes de las regiones variables de las cadenas pesadas o livianas de las inmonuglobulinas de superficie (BCR) y causan la muerte por apoptosis de las células B que los reconocen. Los Sags T se unen a los antígenos mayores de histocompatibilidad de clase II en las células presentadoras de antígenos e interactúan con linfocitos T que expresen una región variable particular de su cadena â. Las células T reactivas mueren después de recibir esta fuerte estimulación. No se conocen intentos de investigar si los Sags son capaces de inducir la muerte por apoptosis de células neoplásicas B o T. En el presente trabajo reportamos que diferentes Sags bacterianos o virales son capaces de inducir la apoptosis de celulas de leucemias/linfomas B y T murinos y humanos. Estudiamos los mecanismos de apoptosis involucrados. En el caso de los linfomas T estudiados encontramos que estaban implicadas tanto la vía extrínseca cuanto la intrínseca e incluso un entrecruzamiento de ambas vías. El tratamiento in vivo de ratones inoculados con diferentes linfomas T fue capaz de incrementar muy significativamente la sobrevida libre de enfermedad. Mientras que el 100% de los ratones no tratados o tratados con un Sag control (no interactuante con el receptor T del linfoma) morían en pocos días, el tratamiento con Sags específcos indujo una sobrevida de entre el 40 al 90% de los mismos. Se investigo la capacidad del Sag B PpL secretado por Finegoldia magna y que interactua con celulas B que expresan la cadena liviana κ para inducir la apoptosis in vivo e in vitro. de celulas de linfomas/leucemias B murinas y humanas κ+. El Sag utilizado indujo la apoptosis en estas celulas utilizando solo la via intrinseca. En resumen, se demostro que los Sags B y T inducen la muerte por apoptosis de cèlulas B y T neoplasicas que los reconocen a traves de su receptor para el antigeno. Se discute la posibilidad del uso de los superantígenos para la implementacion de una nueva herramienta terapeutica que tendria la ventaja de afectar solo un numero discreto de linfocitos sin afectar otros tipos de cèlulas normales.


Superantiges are mostly bacterial or viral proteins that stimulate a large number of normal T or B lymphocytes.. This fact is because - unlike conventional antigens that are recognized by all the variable regions of the receptor of these cells - they only interact with constant regions located within the variable regions of the receptors. B-cell superantigens bind to conserved sites of the VH or VL regions of immunoglobulin molecules outside their complementarity-determining regions causing the apoptosis of normal cognate B cells. T cell Superantigens bind to major histocompatibility complex class II molecules in antigen presenting cells and interact with T cells expressing a particular T cell receptor Vß inducing a strong proliferation/deletion response of the superantigen-reactive T cells. No attempts to investigate whether B or T-cell Sags are able to induce the apoptosis of cognate malignant B or T cells were reported. In the present study we show that bacterial and viral-encoded superantigens induce the apoptosis of murine and human cognate lymphoma T cells both in vitro and in vivo. The extrinsic and the intrinsic patway of apoptosis were found to be involved. Besides, a cross-talk between both pathway was also found. In vivo exposure to bacterial superantigens was able to improve the survival of lymphoma bearing mice. Moreover, the permanent expression of a retroviral encoded superantigen induced the complete remission of an aggressive lymphoma in a high percentage of mice. We also studied the ability of a B-cell superantigen (PpL) secreted by Finegoldia magna, which interacts with κ+ bearing cells, to induce the apoptosis of murine and human κ+ lymphoma B cells both in vitro and in vivo. The involvement of the intrinsic but not the extrinsic patway of apoptosis was clearly demonstrated. In summary, herein we show that B and T superantigens are able to induce the apoptosis of cognate B and T malignat cells.The possibility of a therapeutic use of T and B Superantigens in lymphoma/ leukemia B and T cells is discussed. Their possible use would have the advantage of delete only a discrete number of normal lymphocytes without altering other normal cell types.


Subject(s)
Humans , Lymphocytes/immunology , Apoptosis , Superantigens/immunology , Flow Cytometry , Neoplasms/therapy
9.
Article in Korean | WPRIM | ID: wpr-139535

ABSTRACT

Atopic dermatitis (AD) is characterized by disturbances in epidermal barrier functions and the hyperactive immune response. Staphylococcus aureus (S. aureus) can be cultured from 90% of AD skin lesions and can exacerbate or contribute to the persistent skin inflammation in AD by secreting toxins with superantigenic properties. Superantigens can induce mast cell (MC) degranulation after penetrating the epidermal barrier. The role of MCs in AD is suggested by the increase in the MC number and MC activation. MCs are activated for degranulation and mediator release by allergens that cross-link IgE molecules or by microbial products. Therefore, MCs may be critically involved in the pathogenesis of AD. However, the understanding mechanisms of MC degranulation by S. aureus in relation to AD have still not been fully elucidated. In this study, we found that live S. aureus or methicillin-resistant S. aureus (MRSA) but not heat-killed bacteria induced MC degranulation. The heat-treatment partially inhibited MC degranulation by conditioned media (CM) of S. aureus or MRSA. The calcium chelator ethylene glycol tetraacetic acid (EGTA) did not block MC degranulation induced by live S. aureus or MRSA, but EGTA-treatment partially inhibited MC degranulation by CM from S. aureus or MRSA. These results suggest that live S. aureus and MRSA can degranulate MCs via direct interaction which may be important role in AD.


Subject(s)
Humans , Allergens , Bacteria , Calcium , Culture Media, Conditioned , Dermatitis, Atopic , Egtazic Acid , Immunoglobulin E , Inflammation , Mast Cells , Methicillin Resistance , Methicillin-Resistant Staphylococcus aureus , Skin , Staphylococcus aureus , Superantigens
10.
Article in Korean | WPRIM | ID: wpr-139538

ABSTRACT

Atopic dermatitis (AD) is characterized by disturbances in epidermal barrier functions and the hyperactive immune response. Staphylococcus aureus (S. aureus) can be cultured from 90% of AD skin lesions and can exacerbate or contribute to the persistent skin inflammation in AD by secreting toxins with superantigenic properties. Superantigens can induce mast cell (MC) degranulation after penetrating the epidermal barrier. The role of MCs in AD is suggested by the increase in the MC number and MC activation. MCs are activated for degranulation and mediator release by allergens that cross-link IgE molecules or by microbial products. Therefore, MCs may be critically involved in the pathogenesis of AD. However, the understanding mechanisms of MC degranulation by S. aureus in relation to AD have still not been fully elucidated. In this study, we found that live S. aureus or methicillin-resistant S. aureus (MRSA) but not heat-killed bacteria induced MC degranulation. The heat-treatment partially inhibited MC degranulation by conditioned media (CM) of S. aureus or MRSA. The calcium chelator ethylene glycol tetraacetic acid (EGTA) did not block MC degranulation induced by live S. aureus or MRSA, but EGTA-treatment partially inhibited MC degranulation by CM from S. aureus or MRSA. These results suggest that live S. aureus and MRSA can degranulate MCs via direct interaction which may be important role in AD.


Subject(s)
Humans , Allergens , Bacteria , Calcium , Culture Media, Conditioned , Dermatitis, Atopic , Egtazic Acid , Immunoglobulin E , Inflammation , Mast Cells , Methicillin Resistance , Methicillin-Resistant Staphylococcus aureus , Skin , Staphylococcus aureus , Superantigens
11.
An. bras. dermatol ; An. bras. dermatol;92(5,supl.1): 8-10, 2017. tab, graf
Article in English | LILACS | ID: biblio-887100

ABSTRACT

Abstract: Acne fulminans is a rare and severe variant of acne. In Brazilian medical journals, cases are infrequently reported, confirming its rarity. We followed five young male patients with this severe variant of cutaneous lesions, accompanied by also severe systemic symptoms: fever, anorexia, weight loss, and arthralgia. All had a good response to corticosteroids (prednisone), but had significant scarring.


Subject(s)
Humans , Male , Adolescent , Young Adult , Acne Vulgaris/complications , Acne Vulgaris/pathology , Arthralgia/etiology , Fever/etiology , Propionibacterium acnes/immunology , Severity of Illness Index , Prednisone/therapeutic use , Acne Vulgaris/drug therapy , Superantigens/immunology , Disease Progression , Glucocorticoids/therapeutic use , Necrosis
12.
J. bras. pneumol ; J. bras. pneumol;42(5): 356-361, Sept.-Oct. 2016. tab
Article in English | LILACS | ID: lil-797949

ABSTRACT

ABSTRACT Objective: To determine the presence of staphylococcal superantigen-specific IgE antibodies and degree of IgE-mediated sensitization, as well as whether or not those are associated with the severity of asthma in adult patients. Methods: This was a cross-sectional study involving outpatients with asthma under treatment at a tertiary care university hospital in the city of Rio de Janeiro, Brazil. Consecutive patients were divided into two groups according to the severity of asthma based on the Global Initiative for Asthma criteria: mild asthma (MA), comprising patients with mild intermittent or persistent asthma; and moderate or severe asthma (MSA). We determined the serum levels of staphylococcal toxin-specific IgE antibodies, comparing the results and performing a statistical analysis. Results: The study included 142 patients: 72 in the MA group (median age = 46 years; 59 females) and 70 in the MSA group (median age = 56 years; 60 females). In the sample as a whole, 62 patients (43.7%) presented positive results for staphylococcal toxin-specific IgE antibodies: staphylococcal enterotoxin A (SEA), in 29 (20.4%); SEB, in 35 (24.6%); SEC, in 33 (23.2%); and toxic shock syndrome toxin (TSST), in 45 (31.7%). The mean serum levels of IgE antibodies to SEA, SEB, SEC, and TSST were 0.96 U/L, 1.09 U/L, 1.21 U/L, and 1.18 U/L, respectively. There were no statistically significant differences between the two groups in terms of the qualitative or quantitative results. Conclusions: Serum IgE antibodies to SEA, SEB, SEC, and TSST were detected in 43.7% of the patients in our sample. However, neither the qualitative nor quantitative results showed a statistically significant association with the clinical severity of asthma.


RESUMO Objetivo: Determinar a presença de anticorpos IgE específicos para superantígenos estafilocócicos e o grau de sensibilização mediada por esses, assim como se esses estão associados à gravidade da asma em pacientes adultos. Métodos: Estudo transversal incluindo asmáticos adultos em acompanhamento ambulatorial em um hospital universitário terciário no Rio de Janeiro (RJ). Os pacientes foram alocados consecutivamente em dois grupos de gravidade da asma segundo critérios da Global Initiative for Asthma: asma leve (AL), com asmáticos leves intermitentes ou persistentes, e asma moderada ou grave (AMG). Foram determinados os níveis séricos de anticorpos IgE antitoxinas estafilocócicas, e os resultados foram comparados por análise estatística. Resultados: Foram incluídos 142 pacientes no estudo: 72 no grupo AL (mediana de idade = 46 anos; 59 do sexo feminino) e 70 do grupo AMG (mediana de idade = 56 anos; 60 do sexo feminino). Na amostra geral, 62 pacientes (43,7%) apresentaram resultados positivos para dosagens de anticorpos IgE antitoxinas estafilocócicas: enterotoxina (TX) A, em 29 (20,4%); TXB, em 35 (24,6%); TXC, em 33 (23,2%); e toxic shock syndrome toxin (TSST), em 45 (31,7%). As médias das dosagens séricas de anticorpos IgE específicos anti-TXA, TXB, TXC e TSST foram, respectivamente, de 0,96 U/l, 1,09 U/l, 1,21 U/l, e 1,18 U/l. Não houve diferença estatisticamente significativa dos resultados qualitativos ou quantitativos entre os grupos. Conclusões: A presença de anticorpos IgE séricos anti-TXA, TXB, TXC e TSST, foi detectada em 43,7% nessa amostra de pacientes, mas não houve associação estatisticamente significativa entre seus resultados qualitativos ou quantitativos e gravidade clínica da asma.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Asthma/immunology , Immunoglobulin E/analysis , Severity of Illness Index , Staphylococcus aureus/immunology , Superantigens/immunology , Cross-Sectional Studies , Immunoglobulin E/immunology , Peak Expiratory Flow Rate/immunology
13.
Colomb. med ; 47(1): 15-20, Jan.-Mar. 2016.
Article in English | LILACS | ID: lil-783533

ABSTRACT

Introduction: Staphylococcus aureus is a pathogen that causes food poisoning as well as hospital and community acquired infections. Objective: Establish the profile of superantigen genes among hospital isolates in relation to clinical specimen type, susceptibility to antibiotics and hospital or community acquisition. Methods: Eighty one isolates obtained from patients at Colombian hospital, were classified by antimicrobial susceptibility, specimen type and hospital or community acquired. The PCR uniplex and multiplex was used for detection of 22 superantigen genes (18 enterotoxins, tsst-1 and three exfoliative toxins). Results: Ninety five point one percent of isolates harbored one or more of the genes with an average of 5.6 genes. Prevalence of individual genes was variable and the most prevalent was seg (51.9%). Thirty nine genotypes were obtained, and the genotype gimnou (complete egc cluster) was the most prevalent alone (16.0%) and in association with other genes (13.6%). The correlation between presence of superantigens and clinical specimen or antimicrobial susceptibility showed no significant difference. But there was significant difference between presence of superantigens and the origin of the isolates, hospital or community acquired (p= 0.049). Conclusions: The results show the variability of the superantigen genes profile in hospital isolates and shows no conclusive relationship with the clinical sample type and antimicrobial susceptibility, but there was correlation with community and hospital isolates. The analysis of the interplay between virulence, epidemic and antibiotic resistance of bacterial populations is needed to predict the future of infectious diseases.


Introducción: Staphylococcus aureus, es un patógeno que causa intoxicación alimentaria e infecciones hospitalarias y comunitarias. Objetivo: Establecer el perfil de genes de superantígenos en aislamientos hospitalarios correlacionándolos con el tipo de muestra clínica, susceptibilidad antimicrobiana y origen hospitalario o comunitario. Métodos: Se analizaron 81 aislamientos de S. aureus de pacientes de un hospital colombiano. Fueron clasificadas por susceptibilidad antimicrobiana, tipo de muestra clínica y origen hospitalario o comunitario. Se detectó por PCR individual y múltiple 22 genes de superantígenos (18 enterotoxinas, una toxina del choque tóxico-1 y tres toxinas exfoliativas). Resultados: El 95.1% albergaban uno o más genes de superantígenos con un promedio de 5.6 genes. La prevalencia individual fue variable y el gen con mayor prevalencia fue seg (51.9%). Se obtuvieron 39 genotipos, y el genotipo gimnou (cluster egc completo) fue el de mayor frecuencia (16.0%) y asociado con otros genes (13.6%). La correlación de superantígenos frente a tipo de muestra clínica y susceptibilidad antimicrobiana no mostró diferencia estadística significativa, pero hubo diferencia significativa con el tipo de aislamiento hospitalario o comunitario (p= 0.049). Conclusiones: Los resultados muestran la diversidad genética en los aislados hospitalarios respecto a la presencia de superantígenos y no muestra una relación concluyente con el tipo de muestra clínica y susceptibilidad antimicrobiana pero sí con origen de los aislamientos comunitarios y hospitalarios. Un análisis de la interrelación entre la virulencia, epidemicidad y resistencia antimicrobiana de las poblaciones bacterianas es necesario para predecir el futuro de las enfermedades infecciosas.


Subject(s)
Adult , Female , Humans , Male , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/immunology , Superantigens/genetics , Drug Resistance, Bacterial/genetics , Staphylococcus aureus/genetics , Staphylococcus aureus/pathogenicity , Virulence , Base Sequence , Cross Infection/microbiology , Prospective Studies , DNA Primers , Community-Acquired Infections/microbiology , Nucleic Acid Amplification Techniques
14.
Infectio ; 19(3): 109-114, Sept.-Dec. 2015. tab
Article in Spanish | LILACS, COLNAL | ID: lil-751180

ABSTRACT

Objetivo: Establecer la correlacion entre la deteccion de superantigenos (MSSA) y la susceptibilidad a oxacilina de Staphylococcus aureus (MRSA) en aislamientos hospitalarios. Materiales y métodos: En 81 aislamientos de Staphylococcus aureus de origen hospitalario, la susceptibilidad a oxacilina se establecio por un sistema automatizado y la deteccion de 22 genes de superantigenos fue realizada mediante PCR individual y multiple. Resultados: La MRSA fue del 38,3%. Todos los aislamientos MRSA portaban uno o mas genes de superantigenos; y el 92% de MSSA. El numero de genotipos fue variable, pero un hallazgo relevante fue que el cluster egc solo fue detectado en MRSA (48,4%). Los genes no clasicos mas detectados en MRSA fueron sem (53,1%) y seg (28,4%); y sem (37%) y seq (30,9%) para MSSA. El gen sec clasico (13,6%) fue mas prevalente en MSSA, y en MRSA, los clasicos fueron de muy baja frecuencia. Para todos los genes, los genes seg , sej , sen , seo y seq mostraron diferencias estadisticamente significativas entre los aislamientos MRSA y MSSA. Conclusión: Este estudio no permitio sacar conclusiones concluyentes para establecer la relacion entre la deteccion de superantigenos y la susceptibilidad a oxacilina (MRSA vs. MSSA). Aunque, el numero de genotipos fue variable, la presencia del cluster egc solamente en aislamientos MRSA es un hallazgo interesante, y en posteriores estudios se podria determinar la importancia del cluster egc.


Objective: To establish the relationship between the detection of superantigens (MSSA) and Staphylococcus aureus resistance to oxacillin in hospital isolates. Material and methods: In 81 isolates of Staphylococcus aureus of hospital origin, an oxacillin susceptibility test was performed by an automated system and 22 superantigenic genes were obtained using single and multiplex PCR. Results: The MRSA was 38.3%. All MRSA isolates carried one or more genes for superantigens and 92.0% of MSSA. The numbers of genotypes for the 2 groups were variable, but the most important finding was that the egc cluster was detected only in MRSA (48.4%). The non-classic genes more often detected in MRSA were sem (53.1%) and seg (28.4%); in MSSA they were sem (37.0%) and seq (30.9%). The gen classic sec (13.6%) was more prevalent in MSSA and in MRSA; the classic genes were very low in frequency. For all genes, the genes: seg , sej , sen , seo and seq showed statistically significant differences between MRSA and MSSA isolates. Conclusion: This study did not reveal a clear relationship between the detection of superantigens and oxacillin susceptibility (MRSA vs. MSSA). Although the number of genotypes varied, the presence of egc cluster only in the MRSA isolate was an important finding. Further studies are needed to establish the importance of the egc cluster.


Subject(s)
Humans , Staphylococcal Infections , Oxacillin , Cross Infection , Methicillin Resistance , Superantigens , Enterotoxins
15.
EJMM-Egyptian Journal of Medical Microbiology [The]. 2015; 24 (4): 1-8
in English | IMEMR | ID: emr-175716

ABSTRACT

Background: Eczematous skin of atopic dermatitis [AD] is highly susceptible to infection and colonization by Staphylococcus aureus and the superantigen toxins can worsen the condition


Objectives: To assess the colonization of Egyptian pediatric AD patients with S. aureus and to characterize the superantigen gene profile of isolates in relation to severity and to presence of multiple drug resistant [MDR] strains


Methodology: The study included 53 AD pediatric patients and 45 controls. Severity of AD was assessed by scoring atopic dermatitis [SCORAD] index. Swabs were collected to isolate S. aureus. Isolates were subjected to multiplex PCR reactions for detection of six superantigen genes and to antimicrobial susceptibility tests by disc diffusion method


Results: Colonization with S. aureus was significantly higher [P < 0.0001] in AD children compared to controls and was significantly associated [P= 0.001] with severity. Superantigen genes were detected in 30.1% of isolates. The most prevalent genes were sea [64.5%], seb [32.3%], sec [6.5%] and tsst-1 [3.2%]. Multidrug resistance was found in 63.1% of strains. Severity of AD was significantly higher with strains harboring superantigen genes [P=0.04] and with MDR strains [P=0.0002]. Among methicillin resistant S. aureus [MRSA], seb was the most prevalent superantigen gene [37.5%], while sea was most prevalent in methicillin-susceptible S. aureus [MSSA] [20%], MDR [23.1%] and non MDR isolates [13.2%]


Conclusion: Superantigen genes and multidrug resistance are common in S. aureus colonizing AD patients and are associated with severity. More attention should be paid at performing antimicrobial susceptibility testing before antibiotic therapy


Subject(s)
Adolescent , Child , Humans , Staphylococcus aureus/isolation & purification , Superantigens/immunology , Transcriptome , Colony Count, Microbial , Case-Control Studies , Drug Resistance, Multiple, Bacterial
16.
EJMM-Egyptian Journal of Medical Microbiology [The]. 2015; 24 (4): 105-111
in English | IMEMR | ID: emr-175729

ABSTRACT

Staphylococcal enterotoxins can act as allergens and stimulate production of specific IgE with subsequent development of allergic manifestations. In addition, they act as superantigens that induce cytokine secretion with more increase of IL-4/IFN-gamma ratio causing augmentation of allergic reactions. Herein we tested the hypothesis that exposure to Staphylococccus aureus and its enterotoxins induces immunological changes that contribute to the pathogenesis of allergic rhinitis and bronchial asthma. This study included 45 air way allergic patients, and 45 controls. The following were done: intradermal allergy skin testing, invitro effect of staphylococcal enterotoxin B [SEB] on cytokine secretion from separated peripheral blood mononuclear cells, detection of nasal carriage of S. aureus and its enterotoxin production by ELISA and serum levels of total IgE, SEB-specific IgE and eosinophil cationic protein [ECP] were measured. We found that nasal carriage of enterotoxin producing S. aureus in allergic patients was significantly higher than in control. Blood eosinophilia, total IgEand ECP levels were significantly higher in S. aureus nasal carrier than non-carrier patients. On in vitro exposure of PBMCs to [SEB], IFN-gamma secretion was significantly less in patients than control and IL-4 secretion was significantly more in patients than control. SEB-specific IgE was detected in 15.6% of patients and not detected in control. There was a significant positive correlation between SEB-specific IgE level in patients and markers of severity of allergic reaction including blood eosinophilia, ECP and total IgE level. This study suggests that nasal carriage of enterotoxin producing S. aureus has a potential role in the development and severity of allergic airway diseases


Subject(s)
Adult , Adolescent , Humans , Asthma/etiology , Staphylococcus aureus/immunology , Staphylococcus aureus/pathogenicity , Staphylococcus aureus/isolation & purification , Enterotoxins , Superantigens
17.
Article in English | WPRIM | ID: wpr-34570

ABSTRACT

BACKGROUND: Burn wounds lack normal barriers that protect against pathogenic bacteria, and burn patients are easily colonized and infected by Staphylococcus aureus. Toxic shock syndrome (TSS) is a rare but fatal disease caused by S. aureus. A lack of detectable antibodies to TSS toxin-1 (TSST-1) in serum indicates susceptibility to TSS. METHODS: A total of 207 patients (169 men and 38 women; median age, 42.5 yr) admitted to a burn center in Korea were enrolled in this study. The serum antibody titer to TSST-1 was measured by sandwich ELISA. S. aureus isolates from the patients' nasal swab culture were tested for TSST-1 toxin production by PCR-based detection of the TSST-1 toxin gene. RESULTS: One hundred seventy-four (84.1%) patients showed positive results for antibody against TSST-1. All patients aged > or =61 yr (n=28) and <26 months (n=7) were positive for the anti-TSST-1 antibody. S. aureus was isolated from 70 patients (33.8%), and 58.6% of the isolates were methicillin resistant. Seventeen patients were colonized with TSST-1-producing S. aureus. The antibody positivity in these 17 carriers was 88.2%, and the positivity in the non-carriers was 83.7%. CONCLUSIONS: Most burn patients had antibody to TSST-1, and nasal colonization with TSST-1-producing S. aureus was associated with positive titers of anti-TSST-1 antibody. Additionally, patients with negative titers of anti-TSST-1 antibody might be susceptible to TSS.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult , Antibodies, Bacterial/blood , Bacterial Toxins/genetics , Burns/blood , Enterotoxins/genetics , Enzyme-Linked Immunosorbent Assay , Nasal Cavity/microbiology , Polymerase Chain Reaction , Prevalence , Staphylococcal Infections/epidemiology , Staphylococcus aureus/isolation & purification , Superantigens/genetics
18.
Zhonghua Yu Fang Yi Xue Za Zhi ; (12): 988-992, 2015.
Article in Chinese | WPRIM | ID: wpr-296649

ABSTRACT

<p><b>OBJECTIVE</b>To investigate superantigen gene profiles of group A Streptococcus (GAS) isolated in Beijing pediatric patients in 2014, and to explore the the relationship between superantigen gene profiles with emm types, and GAS infections with diseases.</p><p><b>METHODS</b>A total of 259 GAS strains were isolated from pediatric patients clinically who diagnosed with scarlet fever and pharyngitis from 36 hospitals in Beijing from May to July, 2014.The Superantigens genes of strains were performed by Real-time PCR (speA, speB, speC, speF, speG, speH, speI, speJ, speK, speL, speM, smeZ, ssa). PCR amplification of GAS strain M protein N gene segments were carried ort; products after sequencing comparison were analyzed to determine the GAS types of emm. The differences in distributions of superantigen genes and emm types of GAS isolates were compared between subgroups.</p><p><b>RESULTS</b>Among the 259 GAS strains, the detection rates of 13 superantigens were as the following: speA 48.6% (126), speB 99.2% (257), speC 99.2% (257), speF 98.8% (256), speG 98.5% (255), speH 43.6% (113), speI 46.3% (120), speJ 49.0% (127), smeZ 99.2% (257) and ssa 98.5% (255), respectively, however, speK, speL, and speM were not found. Eleven superantigen gene profiles in all were observed (A-K). The percentage of emm1 strains harbored spe A and speJ were 94.2% (113/120), 95.0% (114/120), respectively, which were significantly higher than those of emm12 strains (5.6% (7/124), 5.6% (7/124), respectively; χ(2) = 191.20, 194.80, P < 0.001). The percentage of emm12 strains harbored speH and speI were 83.9% (104/124), 88.7% (110/124), respectively, which were significantly higher than those of emm1 strains (3.3% (4/120), 4.2% (5/120), respectively; χ(2) = 160.30, 174.90, P < 0.001).The superantigen genotypes of GAS strains and emm types, which were isolated from scarlet fever and pharyngitis cases, were not significant different (P > 0.05).</p><p><b>CONCLUSION</b>The GSA strains isolated in Beijing pediatric patients in 2014, the relevance ratio of speB, speC, speF, smeZ, speG, ssa were higher than others, while speK, speL, and speM were no detected in any GAS strains. The superantigen genes appeared to be associated with the emm type. Furthermore, emm type distribution and superantigen genes were not different between scalet fever and pharyngitis.</p>


Subject(s)
Child , Humans , Antigens, Bacterial , Genetics , Beijing , Genotype , Real-Time Polymerase Chain Reaction , Streptococcal Infections , Microbiology , Streptococcus pyogenes , Genetics , Allergy and Immunology , Superantigens , Genetics
19.
Journal of Rhinology ; : 1-5, 2015.
Article in Korean | WPRIM | ID: wpr-24737

ABSTRACT

Systemic corticosteroids currently represent the most effective treatment for chronic rhinosinusitis with nasal polyps (CRSwNP), but their long-term use is constrained due to their detrimental side effects. Until recently, development of novel drugs for CRSwNP has been difficult partly due to the absence of a standard animal model of CRSwNP. Exotoxins of Staphylococcus aureus such as staphylococcal enterotoxin B (SEB), are well-known superantigens which can induce a strong immune response; there have been many studies on the association of staphylococcal enterotoxins and development of CRSwNP over the past two decades. Based on previous studies, we invented a mouse model of CRSwNP using SEB. Herein, we explain the protocol development for the mouse model, as well as identify histological and immunological similarities between this mouse model and humans. Furthermore, we describe a study that analyzed the risk factors for CRSwNP such as smoking, and also elaborate on a series of studies that searched for new potential drugs for CRSwNP, including resveratrol, anti-periostin antibody, topical hypoxia-inducible factors, and topical cyclosporine. Based on preceding studies, we have concluded that this mouse model might be a useful tool to investigate the pathophysiology and development of novel drugs for CRSwNP.


Subject(s)
Animals , Humans , Mice , Adrenal Cortex Hormones , Cyclosporine , Enterotoxins , Exotoxins , Models, Animal , Nasal Polyps , Risk Factors , Smoke , Smoking , Staphylococcus aureus , Superantigens
20.
Article in English | WPRIM | ID: wpr-85012

ABSTRACT

Protein tyrosine phosphatase-22 (PTPN22) gene encodes lymphoid-specific tyrosine phosphatase (Lyp), an inhibitor of T cell activation. A polymorphism of the PTPN22 gene has been found to be associated with chronic urticaria (CU). We investigated the associations between PTPN22 gene polymorphisms and CU characteristics, including serum specific IgE antibodies response to toxic shock syndrome toxin-1 (TSST-1) and staphylococcal enterotoxin A (SEA). CU patients (n=409) and normal healthy controls (n=388) were enrolled in the present study. Serum specific IgE to TSST-1 and SEA were measured by ImmunoCAP(R). Five PTPN22 single nucleotide polymorphisms, -1123G>C, 1858C>T, 13145A>G, 14943C>T, and 20628A>G, were genotyped. There were no significant differences in genotype or haplotype frequencies of these polymorphisms between the 2 groups. CU patients carrying the GG genotype at 20628A>G (P=0.035) or haplotype 3 [GGG] (P=0.047) had a significantly higher prevalence of serum specific IgE to TSST-1 compared to non-carriers. Similarly, CT/TT genotype at 14943C>T had a significantly higher prevalence of serum specific IgE to SEA (P=0.045). The findings suggest that the PTPN22 gene polymorphisms at 20628A>G and 14943C>T may enhance serum specific IgE responses to TSST-1 and SEA, which may contribute to CU pathogenesis.


Subject(s)
Humans , Antibodies , Enterotoxins , Genotype , Haplotypes , Immunoglobulin E , Polymorphism, Single Nucleotide , Prevalence , Shock, Septic , Superantigens , Tyrosine , Urticaria
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