首页> 外文OA文献 >Protein Expression Profiling of Coccidioides posadasii by Two-Dimensional Differential In-Gel Electrophoresis and Evaluation of a Newly Recognized Peroxisomal Matrix Protein as a Recombinant Vaccine Candidate
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Protein Expression Profiling of Coccidioides posadasii by Two-Dimensional Differential In-Gel Electrophoresis and Evaluation of a Newly Recognized Peroxisomal Matrix Protein as a Recombinant Vaccine Candidate

机译:二维球状差异凝胶电泳分析球孢子虫的蛋白质表达谱并评估新鉴定的过氧化物酶体基质蛋白作为重组疫苗的候选对象

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摘要

Coccidioides posadasii and Coccidioides immitis are dimorphic, soil-dwelling pathogenic ascomycetes endemic to the southwestern United States. Infection can result from inhalation of a very few arthroconidia, but following natural infection, long-lived immunity is the norm. Previous work in the field has shown that spherule-derived vaccines afford more protection than those from mycelia. We have used two-dimensional differential in-gel electrophoresis coupled with nano-high-performance liquid chromatography-tandem mass spectrometry to directly assess both absolute abundance and differential expression of proteins in the spherule and the mycelial phases of C. posadasii with the intent to identify potential vaccine candidates. Peptides derived from 40 protein spots were analyzed and a probable identity was assigned to each. One spherule-abundant protein, identified as Pmp1, showed homology to allergens from Aspergillus fumigatus and other fungi, all of which exhibit similarity to yeast thiol peroxidases. Recombinant Pmp1 was reactive with serum from individuals with both acute and protracted disease, and evoked protection in two murine models of infection with C. posadasii. These results demonstrate the utility of proteomic analysis as a point of discovery for protective antigens for possible inclusion in a vaccine candidate to prevent coccidioidomycosis.
机译:球孢子虫和球孢子虫是美国西南部特有的二态性,土生病原性子囊菌。吸入很少的关节炎可以导致感染,但是自然感染后,长寿命的免疫才是常态。该领域以前的工作表明,来自球菌的疫苗比来自菌丝体的疫苗提供更多的保护。我们已使用二维差分凝胶内电泳与纳米高效液相色谱-串联质谱联用,直接评估了波氏梭菌的球状体和菌丝体相中蛋白质的绝对丰度和差异表达,目的是确定潜在的疫苗候选者。分析了来自40个蛋白斑点的肽,并为每个肽分配了可能的同一性。一种被识别为Pmp1的富球蛋白与烟曲霉和其他真菌的过敏原具有同源性,所有这些都与酵母硫醇过氧化物酶具有相似性。重组Pmp1与患有急性和长期疾病的个体的血清反应,并且在两种感染波氏梭菌的鼠模型中引起保护。这些结果证明了蛋白质组学分析的实用性,作为发现保护性抗原的一个可能点,该保护性抗原可能包含在候选疫苗中以预防球孢子菌病。

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