It has been proposed that activation of Toll-like receptors （TLRs） plays crucial roles in the polarization of adaptive immune responses. A synthetic Toll-like receptor 2 （TLR2） ligand, Pam3CSK4, has been reported ...It has been proposed that activation of Toll-like receptors （TLRs） plays crucial roles in the polarization of adaptive immune responses. A synthetic Toll-like receptor 2 （TLR2） ligand, Pam3CSK4, has been reported to modulate the balance of Th1/Th2 responses. We evaluated the modulation effect of Pam3CSK4 on allergic immune response in a mouse rhinitis model sensitized to house dust mite allergen （HDM）. Mice were sensitized and challenged with Dermatophagoidesfarinae allergen （Der f）, and then the allergic mice were treated by Pam3CSK4. Nasal allergic symptoms and eosinophils were scored. Der f-specific cytokine responses were examined in the splenocytes and bronchoalveolar lavage fluid （BALF）. Serum level of total IgE was also detected. After establishing a mouse allergic rhinitis model with HDM, we have showed that Pam3CSK4 treatment not only ameliorated the nasal allergic symptoms remarkably but also decreased the eosinophils and total inflammation cells in BALF significantly. Analysis of cytokine profile found that IFN-7 released from either BALF or stimulated splenocytes increased markedly in Pam3CSK4-treated mice, while IL-13 decreased significantly. Moreover, serum level of total IgE was significantly lower in Pam3CSK4-treated mice than in the untreated. Thus, in an allergic rhinitis mouse model developed with HDM, Pam3CSK4 was shown to exhibit an antiallergic effect, indicating its potential application in allergic diseases.展开更多
Background Haikou locates in tropical island with unique mite allergens levels in Haikou, and to investigate the prevalence of between house dust mites. propagation. The aim of this stuy is to determine mite mite spec...Background Haikou locates in tropical island with unique mite allergens levels in Haikou, and to investigate the prevalence of between house dust mites. propagation. The aim of this stuy is to determine mite mite specific IgE-sensitization and IgE cross-reactivity Methods Allergen and antigen concentrations against six mite species were tested by enzyme-linked immunosorbent assay （ELISA）. Specific IgE concentrations and cross-inhibitions were measured with ADVIA Centaur . Results Allergen or antigen Dermatophagoides pteronyssinus （Der p 1 ）, Blomia tropicalis （BIot ） and Tyrophagus putrescentia （Tyr p） were detected in dust samples. Dermatophagoides farinae （Der f 1 ）, Lepidoglyphus destructor （Lep d 2）, and Acarus siro （Aca s ） were found in very few samples. Specific IgE tests showed high prevalence of sensitizations against all tested mites with high IgE levels to Der p, Der f, and BIot. Storage mites, BIo t, Tyr p, Lep d, and Aca s, could inhibit Der p from 0 to 50%. Storage mites could inhibit Der f between 30% and 100%. Der p IgE could be inhibited by Der f with up to 90%, and vice versa. Der p could inhibit BIot from 40% to 80%. BIot was able to fully inhibit IgE binding to Lep d, Tyr p, and Aca s compared to partial inhibition by Der p. Conclusions Der p is the dominating mite and has the highest specific IgE prevalence among asthmatic children. BIot represents an important source of storage mite sensitization and some patients may be independently sensitized to both Der p and BIot. High prevalence of sensitization to Der f may be due to IgE-mediated cross-reactivity with Der p and BIo t.展开更多
Background Cockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of I...Background Cockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of IgE between cockroach and house dust mite allergen in Chinese patients.Methods The cockroach sensitization pattern was based on a skin prick test （SPT） obtained from a national multicenter prevalence study, in which 6304 patients from 25 allergy centers across China participated. Factors, including different regions of China, age, gender and the correlations between the American and German cockroaches and house dust mite Der p were investigated. Eighteen out of 1236 clinical sera from south China were selected to perform the cross-inhibition assay between house dust mites and cockroaches.Results Totally 25.7% of patients were SPT positive to the American cockroach （Periplaneta Americana, Per a） and 18.7% SPT positive to the German cockroach （Blattella germanica, Bla g）. The prevalence of positive cockroach SPT was higher in southern than in northern China, higher in adults than in children, and higher in males than in females.Patients had relatively low levels of cockroach SPT reactions, mainly class 1 or 2. Of the SPT positive cockroach patients,88% were also SPT positive to house dust mite （Dermatophagoides pteronyssinus, Der p）. An IgE cross-inhibition study confirmed that Der P sensitization could cause false positive SPT reactions against cockroach.Conclusions A relatively high prevalence of cockroach sensitivity was found in mainland of China. However, a cross-inhibition study showed that only a small number of patients appear to have Bla g and/or Per a as primary sensitizing source. The importance of cockroaches as a risk factor for sensitization and triggers of allergic symptoms in mainland of China needs to be further investigated.展开更多
基于巴斯德毕赤酵母表达系统探讨了屋尘螨过敏原Der p 1的重组表达与纯化。首先,采用毕赤酵母GS115表达密码子优化的全长编码基因PreProDer p 1,其产量可达100 mg/L,进一步共表达分子伴侣实现产量提高到140 mg/L,并实现3 L反应器发酵产...基于巴斯德毕赤酵母表达系统探讨了屋尘螨过敏原Der p 1的重组表达与纯化。首先,采用毕赤酵母GS115表达密码子优化的全长编码基因PreProDer p 1,其产量可达100 mg/L,进一步共表达分子伴侣实现产量提高到140 mg/L,并实现3 L反应器发酵产量提高到1 g/L。其次,对上述重组蛋白发酵液分别使用阳离子交换层析及亲和层析进行了纯化工艺优化,目的蛋白得率达到60.7%。本研究为后续PreProDre p 1诊断试剂盆的开发提供了参考。展开更多
Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samp...Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samples were collected from nine cities. Major allergens Der p 1 and Der f I from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.Results HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from 〈0.006 to 9.2 μg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.Conclusion HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.展开更多
过敏原的硝基化会引起其致敏潜能的增强,进而带来更大的致敏性健康风险.过敏原蛋白质通常含有多个酪氨酸硝基化位点,分析过敏原硝基化的位点选择性是探究硝基化对过敏原致敏性影响的重要基础.本文以尘螨过敏原为研究对象,建立了基于超...过敏原的硝基化会引起其致敏潜能的增强,进而带来更大的致敏性健康风险.过敏原蛋白质通常含有多个酪氨酸硝基化位点,分析过敏原硝基化的位点选择性是探究硝基化对过敏原致敏性影响的重要基础.本文以尘螨过敏原为研究对象,建立了基于超高效液相色谱-串联质谱(UPLC-MS/MS)同时定量分析3种尘螨过敏原(Der f 1、Der p 1和Der p 2)的13个酪氨酸位点硝基化程度的方法,并应用于分析3种尘螨过敏原在过氧亚硝酸盐硝基化作用下的位点选择性.结果表明,3种尘螨过敏原均发生了位点特异性的硝基化,Y195、Y37和Y92分别为Der f 1、Der p 1和Der p 2中反应活性最高的硝基化位点.尘螨过敏原位点选择性的硝基化表明,在评价硝基化尘螨过敏原的致敏性变化时应当考虑其位点特异性的硝基化状况.展开更多
目的:分析粉尘螨过敏原Der f 38的理化性质及结构特征,并综合生物信息学工具预测其免疫优势B、T细胞表位。方法:从世界卫生组织/国际免疫学联合会的过敏原数据库中获取Der f 38的氨基酸序列信息,利用ProtParam对Der f 38的理化性质特征...目的:分析粉尘螨过敏原Der f 38的理化性质及结构特征,并综合生物信息学工具预测其免疫优势B、T细胞表位。方法:从世界卫生组织/国际免疫学联合会的过敏原数据库中获取Der f 38的氨基酸序列信息,利用ProtParam对Der f 38的理化性质特征进行分析。通过SWISS-MODEL、AlphaFold2及GalaxyRefine对Der f 38的三维结构模型进行综合构建、优化得到最佳的蛋白模型。利用DNAStar Protean、Bepipred 2.0、ElliPro、DiscoTop、TepiTool进行Der f 38蛋白的B细胞线性、构象型表位以及T细胞表位的综合预测。结果:Der f 38为分子量13.99kDa且等电点为9.08,总平均亲水性为-0.337,不稳定指数为31.79。通过多种生物信息学工具综合分析,共得到4个优势B细胞线性表位、8个B细胞构象表位和8个T细胞表位。结论:Der f 38属于碱性蛋白,具有一定的亲水性和稳定性,含有多个免疫优势的B细胞和T细胞抗原表位,为进一步针对该蛋白的疫苗和过敏原特异性免疫疗法的设计与开发提供理论参考。展开更多
基金Project (No. 39800135) supported by the National Natural ScienceFoundation of China
文摘It has been proposed that activation of Toll-like receptors （TLRs） plays crucial roles in the polarization of adaptive immune responses. A synthetic Toll-like receptor 2 （TLR2） ligand, Pam3CSK4, has been reported to modulate the balance of Th1/Th2 responses. We evaluated the modulation effect of Pam3CSK4 on allergic immune response in a mouse rhinitis model sensitized to house dust mite allergen （HDM）. Mice were sensitized and challenged with Dermatophagoidesfarinae allergen （Der f）, and then the allergic mice were treated by Pam3CSK4. Nasal allergic symptoms and eosinophils were scored. Der f-specific cytokine responses were examined in the splenocytes and bronchoalveolar lavage fluid （BALF）. Serum level of total IgE was also detected. After establishing a mouse allergic rhinitis model with HDM, we have showed that Pam3CSK4 treatment not only ameliorated the nasal allergic symptoms remarkably but also decreased the eosinophils and total inflammation cells in BALF significantly. Analysis of cytokine profile found that IFN-7 released from either BALF or stimulated splenocytes increased markedly in Pam3CSK4-treated mice, while IL-13 decreased significantly. Moreover, serum level of total IgE was significantly lower in Pam3CSK4-treated mice than in the untreated. Thus, in an allergic rhinitis mouse model developed with HDM, Pam3CSK4 was shown to exhibit an antiallergic effect, indicating its potential application in allergic diseases.
基金The project was partly supported by a grant from Natural Science Foundation of Hainan Province, China （No. 808206）.
文摘Background Haikou locates in tropical island with unique mite allergens levels in Haikou, and to investigate the prevalence of between house dust mites. propagation. The aim of this stuy is to determine mite mite specific IgE-sensitization and IgE cross-reactivity Methods Allergen and antigen concentrations against six mite species were tested by enzyme-linked immunosorbent assay （ELISA）. Specific IgE concentrations and cross-inhibitions were measured with ADVIA Centaur . Results Allergen or antigen Dermatophagoides pteronyssinus （Der p 1 ）, Blomia tropicalis （BIot ） and Tyrophagus putrescentia （Tyr p） were detected in dust samples. Dermatophagoides farinae （Der f 1 ）, Lepidoglyphus destructor （Lep d 2）, and Acarus siro （Aca s ） were found in very few samples. Specific IgE tests showed high prevalence of sensitizations against all tested mites with high IgE levels to Der p, Der f, and BIot. Storage mites, BIo t, Tyr p, Lep d, and Aca s, could inhibit Der p from 0 to 50%. Storage mites could inhibit Der f between 30% and 100%. Der p IgE could be inhibited by Der f with up to 90%, and vice versa. Der p could inhibit BIot from 40% to 80%. BIot was able to fully inhibit IgE binding to Lep d, Tyr p, and Aca s compared to partial inhibition by Der p. Conclusions Der p is the dominating mite and has the highest specific IgE prevalence among asthmatic children. BIot represents an important source of storage mite sensitization and some patients may be independently sensitized to both Der p and BIot. High prevalence of sensitization to Der f may be due to IgE-mediated cross-reactivity with Der p and BIo t.
文摘Background Cockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of IgE between cockroach and house dust mite allergen in Chinese patients.Methods The cockroach sensitization pattern was based on a skin prick test （SPT） obtained from a national multicenter prevalence study, in which 6304 patients from 25 allergy centers across China participated. Factors, including different regions of China, age, gender and the correlations between the American and German cockroaches and house dust mite Der p were investigated. Eighteen out of 1236 clinical sera from south China were selected to perform the cross-inhibition assay between house dust mites and cockroaches.Results Totally 25.7% of patients were SPT positive to the American cockroach （Periplaneta Americana, Per a） and 18.7% SPT positive to the German cockroach （Blattella germanica, Bla g）. The prevalence of positive cockroach SPT was higher in southern than in northern China, higher in adults than in children, and higher in males than in females.Patients had relatively low levels of cockroach SPT reactions, mainly class 1 or 2. Of the SPT positive cockroach patients,88% were also SPT positive to house dust mite （Dermatophagoides pteronyssinus, Der p）. An IgE cross-inhibition study confirmed that Der P sensitization could cause false positive SPT reactions against cockroach.Conclusions A relatively high prevalence of cockroach sensitivity was found in mainland of China. However, a cross-inhibition study showed that only a small number of patients appear to have Bla g and/or Per a as primary sensitizing source. The importance of cockroaches as a risk factor for sensitization and triggers of allergic symptoms in mainland of China needs to be further investigated.
文摘基于巴斯德毕赤酵母表达系统探讨了屋尘螨过敏原Der p 1的重组表达与纯化。首先,采用毕赤酵母GS115表达密码子优化的全长编码基因PreProDer p 1,其产量可达100 mg/L,进一步共表达分子伴侣实现产量提高到140 mg/L,并实现3 L反应器发酵产量提高到1 g/L。其次,对上述重组蛋白发酵液分别使用阳离子交换层析及亲和层析进行了纯化工艺优化,目的蛋白得率达到60.7%。本研究为后续PreProDre p 1诊断试剂盆的开发提供了参考。
文摘Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samples were collected from nine cities. Major allergens Der p 1 and Der f I from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.Results HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from 〈0.006 to 9.2 μg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.Conclusion HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.
文摘过敏原的硝基化会引起其致敏潜能的增强,进而带来更大的致敏性健康风险.过敏原蛋白质通常含有多个酪氨酸硝基化位点,分析过敏原硝基化的位点选择性是探究硝基化对过敏原致敏性影响的重要基础.本文以尘螨过敏原为研究对象,建立了基于超高效液相色谱-串联质谱(UPLC-MS/MS)同时定量分析3种尘螨过敏原(Der f 1、Der p 1和Der p 2)的13个酪氨酸位点硝基化程度的方法,并应用于分析3种尘螨过敏原在过氧亚硝酸盐硝基化作用下的位点选择性.结果表明,3种尘螨过敏原均发生了位点特异性的硝基化,Y195、Y37和Y92分别为Der f 1、Der p 1和Der p 2中反应活性最高的硝基化位点.尘螨过敏原位点选择性的硝基化表明,在评价硝基化尘螨过敏原的致敏性变化时应当考虑其位点特异性的硝基化状况.
文摘目的:分析粉尘螨过敏原Der f 38的理化性质及结构特征,并综合生物信息学工具预测其免疫优势B、T细胞表位。方法:从世界卫生组织/国际免疫学联合会的过敏原数据库中获取Der f 38的氨基酸序列信息,利用ProtParam对Der f 38的理化性质特征进行分析。通过SWISS-MODEL、AlphaFold2及GalaxyRefine对Der f 38的三维结构模型进行综合构建、优化得到最佳的蛋白模型。利用DNAStar Protean、Bepipred 2.0、ElliPro、DiscoTop、TepiTool进行Der f 38蛋白的B细胞线性、构象型表位以及T细胞表位的综合预测。结果:Der f 38为分子量13.99kDa且等电点为9.08,总平均亲水性为-0.337,不稳定指数为31.79。通过多种生物信息学工具综合分析,共得到4个优势B细胞线性表位、8个B细胞构象表位和8个T细胞表位。结论:Der f 38属于碱性蛋白,具有一定的亲水性和稳定性,含有多个免疫优势的B细胞和T细胞抗原表位,为进一步针对该蛋白的疫苗和过敏原特异性免疫疗法的设计与开发提供理论参考。
