Intervention effect and mechanism of Hippophae rhamnoides oil on glucocorticoid resistance in superantigen-induced atopic dermatitis mice / 中国药房

OBJECTIVE To study the intervention effect of Hippophae rhamnoides oil on glucocorticoid resistance in superantigen-induced atopic dermatitis （AD） mice，and to explore the mechanism of action. METHODS Fifty mice were randomly divided into 5 groups，i.e. normal control group （group A），model group （group B），dexamethasone intervention group （positive control，group C），H. rhamnoides oil intervention group （group D），dexamethasone+H. rhamnoides oil intervention group （group E），with 10 mice in each group. Except for group A，other groups were given 2，4-dinitrochlorobenzene+staphylococcal enterotoxin B to induce the AD mice model. Starting from the 7th day of the experiment，groups C，D and E were given dexamethasone （1.5 mg/kg） and/or H. rhamnoides oil （10 mL/kg） intragastrically，once a day，for 28 consecutive days. After the last medication，the pathomorphological changes of ear tissue were observed by 节作用。E-mail：57667478@qq.com HE staining； the serum levels of immunoglobulin E （IgE） and interleukin 4 （IL-4） were detected by enzyme-linked immunosorbent assay. Positive cell count of glucocorticoid receptor α （GRα） and GRβ in the ear tissue of mice was detected by tyramide signal amplification. The expressions of GRα protein，GRβ protein，and protein kinase B （AKT）/ribosomal protein S6 kinase 1，S6K1 （S6K1） signaling pathway-related proteins were determined by Western blot assay. RESULTS Compared with group B，the skin inflammation in the left ear of the mice was significantly reduced in groups C，D and E，the serum levels of IgE and IL-4 were decreased significantly in groups D and E （P＜ 0.05），while the number of GRα positive cells and GRα protein expression were increased significantly （P＜0.05）； the protein levels of G protein inhibitory subunit 1 （Gαi1），Gαi3，phosphorylated S6K1 （p-S6K1） and phosphorylated AKT （p-AKT） were decreased significantly （P＜0.05）； the number of GRβ positive cells and protein expression of GRβ was decreased significantly in group E（P＜0.05）. Compared with group C，the skin inflammation in the left ear of the mice was almost clear away in group E，the serum levels of IgE and IL-4 were decreased significantly （P＜0.05）； the number of GRα positive cells and GRα protein expression were increased significantly in groups D and E （P＜0.05）； the protein levels of GRβ，Gαi1，p-S6K1 and p-AKT were all decreased significantly in groups D and E（P＜0.05）； and protein level of Gαi3 was decreased significantly in group E （P＜0.05）. CONCLUSIONS H. rhamnoides oil has an intervention effect on superantigen-induced glucocorticoid resistance of AD mice，which may be exerted by inhibition of the Gαi1/3-induced AKT/S6K1 signaling pathway.

Advances in the classification and biological properties of streptococcal superantigens / 中华实用儿科临床杂志

Superantigens are highly mitogenic exotoxins produced by bacteria and viruses.They can bind to the major histocompatibility complex class Ⅱ, stimulate T cell receptor molecules to release a large quantity of proinflammatory cytokines, and activate the immune system in vastly low concentrations.Superantigens have been the most effective T cell mitogen discovered so far.Superantigens play important roles in infection diseases caused by streptococcal pathogens, and they can facilitate pathogen colonization, proliferation, and dissemination.Further research of biological chara-cteristics of streptococcus superantigens is vital to understanding the streptococcus pathogenesis, prevention and treatment of streptococcus infection.In this article, advances in the classification, biological properties, pathogenesis, deve-lopment and application of streptococcal superantigens were reviewed, so as to improve the clinical understanding and provide reference for in-depth research of related diseases.

Distribution characteristics of emm genotype and superantigen of Group A Streptococcus isolated from the pharynx of children with scarlet fever-like erythema in Beijing / 中华实用儿科临床杂志

Objective:To analyze the genetic characteristics of Group A Streptococcus (GAS) isolated from the pharynx of children with scarlet fever-like erythema. Methods:Pharyngeal GAS isolates were collected retrospectively from children with scarlet fever-like erythema admitted in the Department of Dermatology, Beijing Children′s Hospital, Capital Medical University from June 2019 to February 2020.PCR and sequencing were used to detect emm genotyping and superantigen genes( speA, speB, speC, speF, speG, speH, speI, speJ, speK, speL, speM, ssa and smeZ) of the isolates.Data were compared by the chi-square test or Fisher′ s exact test between groups. Results:A total of 147 GAS strains were collected.The main emm genotypes were emm1.0 in 76 strains (51.70%) and emm12.0 in 60 strains (40.82%). Other 7 emm genotypes were all found in less than 5 strains.The detection rate of speF, smeZ, speG, speC, speB and ssa were as high as 100.00%(147/147 strains), 100.00%(147/147 strains), 99.32%(146/147 strains), 95.24%(140/147 strains), 94.56%(139/147 strains) and 92.52%(136/147 strains), respectively. speA, speJ, speI, speH and speM had a low detection rate of 51.70%(76/147 strains), 49.66%(73/147 strains), 32.65%(48/147 strains), 23.81%(35/147 strains) and 4.08%(6/147 strains), respectively.No speK and speL were detected.The frequencies of speA and speJ in emm1.0 strains (65/76 strains, 85.53% and 64/76 strains, 84.21%) were significantly higher than those in emm12.0 strains (4/60 strains, 6.67% and 6/60 strains, 10.00%). However, the frequencies of speH and speI in emm1.0 strains (7/76 strains, 9.21% and 2/76 strains, 2.63%) were significantly lower than those in emm12.0 strains (28/60 strains, 46.67% and 45/60 strains, 75.00%) (all P<0.001). Conclusions:At present, emm1.0 and emm12.0 are the main GAS strains isolated from the throat of children with scarlet fever-like erythema in Beijing, and emm1.0 is more common.There is a correlation between emm genotyping and the superantigen gene profile.Type 1 superantigen gene profile is significantly more frequently detected in emm1.0 strains than in emm12.0 strains.Type 2, 3 and 4 superantigen gene profiles are significantly more frequently detected in emm12.0 strains than in emm1.0 strains.

Amebic Liver Abscess and Kawasaki Disease

Background: Co-occurrence of amebic hepatitis and Kawasaki disease has not beenreported previously. Case characteristics: We describe two children (aged 4 y and 5 y)with Kawasaki disease and coexisting liver abscess. They were treated with intravenousimmunoglobulins with/without percutaneous drainage in combination with amebicidalagents. Outcome: Both the children were completely cured of the amebic hepatitis, andhad normalization and regression of coronaries at follow-up. Message: We report theco-existence of amebic hepatitis with Kawasaki disease.

Study on the super-antigen genes of group A Streptococcus pyogenes strains isolated from patients with scarlet fever and pharyngeal infection, in Beijing, 2015-2017 / 中华流行病学杂志

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 emml and emml2 strains (x2=38.196,P＜0.001;x 2=72.310,P＜0.001).There also appeared significant differences in the frequencies of speA,speH,speI and speJ between emm 1 and emm 12 strains (x2 =146.154,P＜ 0.001;x2=52.31,P＜0.001;x2=58.43,P＜0.001;x2=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.

Study on the super-antigen genes of group A Streptococcus pyogenes strains isolated from patients with scarlet fever and pharyngeal infection, in Beijing, 2015-2017 / 中华流行病学杂志

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 emml and emml2 strains (x2=38.196,P＜0.001;x 2=72.310,P＜0.001).There also appeared significant differences in the frequencies of speA,speH,speI and speJ between emm 1 and emm 12 strains (x2 =146.154,P＜ 0.001;x2=52.31,P＜0.001;x2=58.43,P＜0.001;x2=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.

Frequência de genes codificadores de toxinas em Staphylococcus aureus isolados de leite de tanques expansão comunitários / Frequency of toxin-encoding genes in Staphylococcus aureus isolated from community milk tanks

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)

Effect of Fusion Protein TAP-SSL5 on Atherosclerotic Plaque Formation in ApoE Knockout Mice / 中国药师

Objective: To investigate the effect of fusion protein tick anticoagulant peptide (TAP)-staphylococcus aureus superantigen-like protein 5 (SSL5) on the formation of atherosclerotic plaque in ApoE knockout (ApoE-/-) mice.Methods: Totally 21 male 12-week-old ApoE-/-mice were randomly divided into three groups: TAP-SSL5 (3 mg·kg-1·d-1) group, SSL5 (2 mg·kg-1·d-1) group and the blank control group (pH 7.4 phosphate buffer), ip, qd, for 12 weeks.The changes of body mass were observed.The mice were fed with high cholesterol diet for 12 weeks, and then the levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) in plasma were detected.The aorta of mice was subjected to paraffin section and routine HE staining.The formation of atherosclerotic plaque in the aortic root was analyzed.The distribution of atherosclerotic plaques was observed by oil red O staining of the aorta.Results: Compared with that of the blank control group, the increasement of body weight of TAP-SSL5 group and the level of TC significantly decreased (P <0.001), while TG, HDL-C and LDL-C did not change significantly.The HE staining results showed that the plaque area of root slice in the aorta in TAP-SSL5 group was significantly lower than that in the blank control group (P<0.05).The red O staining of aorta showed that the formation of atherosclerotic plaque in TAP-SSL5 group was significantly smaller than that in the blank control group.Conclusion: TAP-SSL5 can significantly inhibit the formation of atherosclerotic plaques in the arteries of ApoE-/-mice.

The long-term toxicity study of TAP-SSL5 fusion protein / 中国生化药物杂志

Objective To investigate the long-term toxicity of anti-inflammatory and anticoagulant tick anticoagulant peptide (TAP)-staphylococcal superantigen like protein-5 (SSL5) fusion protein in normal rats.MethodsSD rats were intraperitoneally injected with TAP-SSL5 (1mg/kg, every other day) for eight weeks and followed up for one week.The general behavior, weight, blood routine test, blood biochemistry and organ indexe were measured.ResultsOur results showed that there were no significantly difference between the TAP-SSL5 treated rats and the control on general behavior, weight, blood routine test, blood biochemistry, organ indexe and pathology.ConclusionThe fusion protein TAP-SSL5 with little long-term toxicity for rats is proved to be a safe drug.

Study on design and three-dimension structure of a broad-spectrum inhibitory peptide against staphylococcal enterotoxins superantigen / 中国免疫学杂志

Objective:Peptides were designed on the basis of high conservative regions of amino acid sequences and structures of the SEs,three-dimension structure of P72 was constructed.Methods: Bioinformatics analysis softwares such as Vector NTI 10.3, InsightII 2000,Discovery Studio 1.7 were used to analyse and predict the space structure of P72.Results:three-dimensional domains of the peptide P72 from SEA, SEB and SEC were quite similar, Peptide P72 was far away from TCRVβchain and MHC class II molecule.Conclusion:The inhibitory activity of peptide P72 may not due to binding to MHCⅡ and TCRVβchain.The exact mechanism of inhibitory activity of P72 should be explored.

Correlación entre la detección de superantígenos y resistencia a oxacilina en aislamientos hospitalarios de Staphylococcus aureus / Correlation between detection of superantigens and oxacillin resistance of Staphylococcus aureus in hospital isolates

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.

Effect of TAP-SSL5 fusion protein on binding of activated platelets to hu-man lymphocytes / 中国病理生理杂志

AIM: To study the effect of tick anticoagulant peptide-staphylococcal superantigen like protein 5 (TAP-SSL5), an anti-inflammatory and anticoagulant fusion protein , on the binding of activated platelets to human lym-phocytes.METHODS:Human periphery lymphocytes were isolated by magnetic activated cell sorting (MACS).The toxic-ity of TAP-SSL5 on the viability of Jurkat cell was assessed by CCK-8 assay.Flow cytometry was applied to detect the ex-pression of CD162 (PSGL-1) on the Jurkat cells (human peripheral blood leukemia T lymphocyte cell line ) and the inhibi-tory effect of TAP-SSL5 on the binding of mouse anti-human CD162 monoclonal antibody (KPL-1) to Jurkat cells.Platelets were activated by ADP at concentration of 20μmol/L, the binding rates of activated platelets to Jurkat cells or human lym-phocytes were assayed by flow cytometry .RESULTS:The concentration of TAP-SSL5 below 30 mg/L didn’ t affect the vi-ability of Jurkat cells .TAP-SSL5 at 10 mg/L competitively inhibited KPL-1 binding to Jurkat cells .The binding rates of activated platelets to Jurkat cells or lymphocytes were (11.86 ±4.49)% and (8.32 ±1.00)%, respectively, which de-creased to (6.73 ±2.71)%and (5.51 ±0.70)%after the Jurkat cells and lymphocytes were pre-incubated with 10 mg/L TAP-SSL5 (P <0.05).CONCLUSION:TAP-SSL5 binds to PSGL-1 expressed on lymphocyte surface and directly in-hibits the binding of activated platelets to human lymphocytes , which may be one of the anti-inflammatory mechanisms of TAP-SSL5.

Preparation and clinical application of esophagus cancer vaccine / 中国综合临床

Objective To explore the anticancer mechanism of esophageal cancer vaccines and clinical effect.Methods Esophagus cancer cells (Eca109) were cultured and the soluble antigen were extracted from the cells.Esophagus cancer vaccine was constructed with the antigen and superantigen SEC.Lymphocytes were isolated from peripheral blood and then stimulated with the vaccine in vitro.Phenotypes of the cells were checked by FCM and killing activity was tested with cytotoxic assays.One hundred and six early esophageal cancer patients were selected after surgery,who were divided into the observation group of 53 cases with esophageal cancer vaccine therapy and 53 cases in the control group with conventional treatment.Then clinical effect was observated and 3 year follow-up survival rate was observed of the of two groups of patients.Results Proliferation of vaccine stimulated lymphocyte group was the strongest,and high peaked at 72 h (A =0.22),and raise CD8+ T cell populations of CTLs.The killing activity of lymphocyte group stimulated by the vaccine against target cells was significantly higher than that of lymphocyte group ((97.36±2.11) ％) vs.(79.27±5.57) ％,F =38.62,P＜0.01).Three years follow up shows that the survival rates of experiment group were 48.27％ (male) and 45.83％ (female) respectively,and control group were 21.43％ (male) and 24.00％ (female),and the difference was significant (x2 =5.06,6.28,P ＜ 0.05).Conclusion The tumor vaccine constructed with esophagus cancer antigen and superantigen SEC can induce PBMC to activate and proliferate into CD8+ CTL with specific cytotoxicity against the cells which the antigen comes from.The vaccine may raise the survival rate of the patients with Esophagus cancer.

Analysis of the effect of clinical application of vaccine for esophageal cancer / 中国肿瘤临床

Objective:To analyze the effects of the clinical applications of vaccine for esophageal cancer to provide treatment ba-sis. Methods:Tumor-soluble antigen was extracted from clinical samples of esophageal cancer tissue. Vaccine for esophageal cancer was prepared using antigen and superantigen staphylococcal enterotoxin C (SEC). One group of patients with esophageal cancer was treated with vaccine after surgery (treatment group). Another group of patients with esophageal cancer was treated using routine meth-ods (control group). After five years, the patients were followed up to analyze survival. Results:After five years of follow up, the sur-vival rates of treatment and control groups were 173/328 (49.71%) and 67/326 (20.55%), respectively. Survival rates significantly dif-fered between the two groups (P<0.05). Furthermore, the survival rates of the female patients were higher than those of the male pa-tients (P<0.05). In the treatment group, the survival rates of patients with low-differentiation cancer were higher than those of patients with high-differentiation cancer (P<0.05). In the control group, the survival rates of patients with low-differentiation cancer were lower than those of patients with high-differentiation cancer (P<0.05). Conclusion:Tumor vaccine prepared with esophageal cancer antigen and superantigen SEC could increase the survival rate of patients with esophageal cancer, particularly female patients and those with low-differentiation cancer.

Effect of 20 and 22 amino acid on superantigen activity of staphylococcal enterotoxin C2 / 中国免疫学杂志

Objective:To investigate the improved superantigen activity of SEC2(T20L/G22E) compared with recombinant staphylococcal enterotoxins C 2 ( rSEC2 ).Methods: The proliferation of spleen lymphocytes and T-cell subpopulations induced by rSEC2 and SEC2(T20L/G22E) were examined by WST-1 and flow cytometry separately,and the gene expression of cytokines and Vβspecificities were quantified by real-time PCR.Results: WST-1 and Flow cytometry assays showed that the superantigen activity of SEC2(T20L/G22E) was improved due to enhanced T-cell stimulating potency,resulting in massive activation of T-cells,particularly CD4+and CD8+T-cells.Quantitative real-time PCR assay showed that despite similar Vβspecificities induced by rSEC 2 and SEC2 (T20L/G22E),the quantities of activated T-cells bearing specific Vβwere different,and SEC2(T20L/G22E) could stimulate more gene expression of associated cytokines simultaneously.Conclusion: The results strongly suggested that the increased SEC 2 ( T20L/G22 E)-TCR-binding affinity contributed to more T-cells activation and cytokine release ,which elicit powerful immune activition.

Effects of Zn2+and Zn2+-coordinating glutamic acid residues on the superantigenic activities of staph-ylococcal enterotoxin C2 / 中华微生物学和免疫学杂志

Objective To investigate the role of Zn 2+and Zn2+-coordinating glutamic acid residues ( 71Glu and 80Glu ) in the superantigenic activities of staphylococcal enterotoxin C2 ( SEC2 ) .Methods Over-lap PCR was used to amply genes encoding recombinant mutant proteins rSEC 2 ( E71A), rSEC2 (E80A) and rSEC2 (E71A/E80A).The mutant proteins were expressed in E.coli BL21 (DE3) and puri-fied by affinity chromatography .The differences of biological activities between rSEC 2 and its mutants were compared in vitro.The effects of Zn 2+on the superantigenic activities of rSEC 2 were evaluated by analyzing the proliferation of splenic lymphocytes in the presence or absence of ethylenediaminetetraacetic acid ( EDTA) .Results The substitution of glutamic acid residue at position 71 and 80 by alanine residue had no significant effects on the superantigenic activities and the conformational stability of rSEC 2 mutants.How-ever, traces of Zn2+(10μmol/L) could significantly enhance rSEC2-induced proliferation of splenic lympho-cytes, and only certain amount of Zn 2+could completely restore rSEC2-induced proliferation in the presence of EDTA.Conclusion This study indicated that mutations at Zn 2+-coordinating glutamic acid residues had no significant effects on the conformational stability and the superantigenic activities of SEC 2.Zn2+might play a role in regulating the superantigenic activities of SEC 2 through some indirect ways .

The Prevalence of Serum Specific IgE to Superantigens in Asthma and Allergic Rhinitis Patients

Staphylococcus aureus is the most common bacterium present in upper respiratory tract, and the toxins it produced are involved in allergic inflammation pathogenesis. In this study, we investigated the clinical significance of IgE in association with staphylococcal superantigens in allergic asthma with rhinitis (BAwAR) and allergic rhinitis alone (AR). We recruited 100 patients with BAwAR (group I), 100 patients with AR (group II), and 88 healthy controls (group III). Patients were clinically diagnosed by physicians, and were sensitized to house dust mites. Specific IgE antibodies to staphylococcal superantigen A (SEA), B (SEB), and toxic shock syndrome toxin-1 (TSST-1) were measured using the ImmunoCAP system. Other clinical parameters were retrospectively analyzed. All specific IgE antibodies to SEA, SEB, and TSST-1 were detected most frequently in group I (22%, 21%, and 27%), followed by group II (11%, 14%, and 21%) and group III (4.5%, 3.4%, and 2.3%). Absolute values of serum specific IgE to SEA, SEB, and TSST-1 were also significantly higher in group I (0.300+/-1.533 kU/L, 0.663+/-2.933 kU/L, and 0.581+/-1.931 kU/L) and group II (0.502+/-2.011 kU/L, 0.695+/-3.337 kU/L, and 1.067+/-4.688 kU/L) compared to those in group III (0.03+/-0.133 kU/L, 0.03+/-0.14 kU/L, and 0.028+/-0.112 kU/L). The prevalence of serum specific IgE to SEA was significantly higher in group I compared to group II (P=0.025). Blood eosinophil counts were significantly higher in patients with specific IgE to SEA or SEB, and higher serum levels of specific IgE to house dust mites were noted in patients with specific IgE to TSST-1. In conclusion, the present study suggested that IgE responses to staphylococcal superantigens are prevalent in the sera of both BAwAR and AR patients. This may contribute to an augmented IgE response to indoor allergens and eosinophilic inflammation.

Clinical features of elderly chronic urticaria

BACKGROUND/AIMS: Chronic urticaria (CU) is defined as itchy wheals lasting 6 weeks or more. As the aged population increases worldwide, it is essential to identify the specific features of this disease in the elderly population. METHODS: We investigated the prevalence and clinical features of CU in elderly patients. Medical records of 837 CU patients from the outpatient Allergy Clinic of Ajou University Hospital, Korea were analyzed retrospectively. Patients with chronic spontaneous urticaria according to the EAACI/GA2LEN/EDF/WAO guidelines were included. Patients older than 60 years were defined as elderly. RESULTS: Of the 837 patients, 37 (4.5%) were elderly. In elderly versus nonelderly CU patients, the prevalence of atopic dermatitis (AD) was significantly higher (37.8% vs. 21.7%, respectively; p = 0.022), while that of aspirin intolerance was lower (18.9% vs. 43.6%, respectively; p = 0.003) in terms of comorbid conditions. The prevalences of serum specific immunoglobulin E antibodies to staphylococcal enterotoxin A and staphylococcal enterotoxin B were considerably higher in elderly CU patients with AD than in those without AD (37.5% vs. 0%, respectively). CONCLUSIONS: Elderly patients with CU had a higher prevalence of AD. Therefore, there is a need to recognize the existence of AD in elderly CU patients.

A genetic effect of IL-5 receptor alpha polymorphism in patients with aspirin-exacerbated respiratory disease

Persistent eosinophil activation in both the upper and lower airway mucosa is a central feature of aspirin-exacerbated respiratory disease (AERD). Eosinophil activation and survival are profoundly influenced by interleukin 5 (IL-5) and its receptor, IL-5R. In patients susceptible to allergic disorders, IL-5 receptor alpha (IL5RA) polymorphisms have been reported; however, an association with AERD remains unclear. We hypothesize that IL5RA polymorphisms may contribute to eosinophil activation in AERD patients. We recruited 139 AERD patients, 171 aspirin-tolerant asthma patients and 160 normal controls. IL5RA polymorphisms (-5993G>A, -5567C>G and -5091G>A) were genotyped and functional activity of polymorphism was assessed by luciferase reporter assay and electrophoretic mobility shift assay (EMSA). There was no significant difference in the genotype frequency of the three polymorphisms among the three groups. AERD patients carrying the AA genotype at -5993G>A had a significantly higher presence of serum-specific immunoglobulin E (IgE) to staphylococcal enterotoxin A (P=0.008) than those with the GG/GA genotype. In vitro, the -5993A allele had a higher promoter activity compared with the -5993G allele in human mast cell (HMC-1; P=0.030) and human promyelocytic leukemia (HL-60; P=0.013) cells. In EMSA, a -5993A probe produced a specific shifted band than the -5993G had. These findings suggest that a functional polymorphism in IL5RA may contribute to eosinophil and mast cell activation along with specific IgE responses to staphylococcal enterotoxin A in AERD patients.

Células T y sus receptores αß y γδ en biopsias de pacientes con enfermedad periodontal / T cells and its receptords and αß and γδ in biopsies from patients with periodontal disease

El propósito de este estudio es determinar la presencia y localización de las células T y de sus receptores αβ y γδ en biopsias de tejido gingival de pacientes con enfermedad periodontal. Se evaluaron 60 biopsias de 12 pacientes, 4 con diagnostico de periodontitis agresiva, 4 con periodontitis crónica y 4 con gingivitis, las cuales fueron procesados para su análisis histológico, inmunohistoquímico e histomorfometrico. Al analizar los resultados por diagnostico los marcadores que mas predominaron fueron, en Gingivitis CD3, CD8 y TCR γδ en tejido conectivo. En Periodontitis crónica CD3, CD8 y TCR γδ en epitelio oral y CD4 el cual presentó una expresión homogénea en los tejidos analizados. En periodontitis agresiva CD3 y CD8 en epitelio crevicular, con una distribución similar entre CD4 y CD8 tanto en epitelio oral como en tejido conectivo y TCR γδ en conectivo. En cuanto a las cadenas variables del TCR Vβ los más expresados en las diferentes patologías estudiadas fueron el 6.7, 8.1 y 12 a nivel del tejido conectivo. Los estudios sobre la expresión de estas familias parecen indicar que es otra vía de activación a tener en cuenta dentro del modelo de la patogenia de la enfermedad y que debe ser estudiado en modelos longitudinales en pacientes con pérdida de inserción progresiva.

T the purpose of this study is identifying the presence and localization of T cells and their receptor αβ and γδ in biopsies of gingival tissue in patients with periodontal disease. 60 biopsies were evaluated in 12 patients, 4 patients with diagnosis of gingivitis, 4 patients with chronic periodontitis and 4 with aggressive periodontitis, which were processed for the histological, immunohistochemical and histomorphometric analysis. The results by diagnosis showed that in gingivitis the more predominant markers were CD3, CD8 and TCR γδ in connective tissue. In chronic periodontitis the markers with bigger expression were CD3, CD8 and TCR γδ in oral epithelium and CD4 that showed a homogeneous behavior in the analized tissues. In aggressive periodontitis CD3 and CD8 in surcular epithelium, TCR γδ in connective tissue and CD4 and CD8 with a similar distribution in oral epithelium and connective tissue. In relation with variable chains of TCR Vβ, the most predominat in the different diagnosis were 6.7,8.1 and 12 in connective tissue. The investigations about the expression of these families indicate that it can be other important via of activation in the pathogenesis of periodontal disease and it should be study in longitudinal models in patients with progressive loss of attachment level.