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[从阿根廷副结核分枝杆菌亚种菌株生产和评估纯化蛋白衍生物]

[Production and evaluation of a purified protein derivative from an Argentine strain of Mycobacterium avium subsp. paratuberculosis].

作者信息

Gioffré Andrea, Echeverría-Valencia Gabriela, Zumárraga Martín, Morsella Claudia, Mon María L, Viale Mariana, Paolicchi Fernando, Romano María I

机构信息

Instituto de Biotecnología, CICVyA-INTA Castelar, Dr. Nicolás Repetto y De Los Reseros S/N, (1686) Hurlingham, Buenos Aires, Argentina.

出版信息

Rev Argent Microbiol. 2012 Jul-Sep;44(3):155-64.

Abstract

Purified Protein Derivatives (PPDs) are non-defined antigens prepared from mycobacteria cultures. They are usually employed to evaluate the specific cellular immune response both in animals and humans. Bovine and avian PPDs are usually employed as antigens in mycobacterial infections such as tuberculosis and paratuberculosis. Nevertheless, PPD from Mycobacterium avium subsp. paratuberculosis, (PPDj) is neither commonly used nor frequently available. However, PPD from Mycobacterium avium subsp. avium is in fact used. We aimed to obtain and evaluate the performance of a PPDj from a local isolate of MAP using the ãInterferon-release assay. The stimulation of ãInterferon-release was significantly different between infected and control cattle when this antigen, named PPDj-IB, was used. Stimulation in the infected animals was similar with both antigens (PPDa and PPDj-IB). However, some animals were positively stimulated with PPDj-IB and not with PPDa. We demonstrated by Western blot that two antigenic molecules, lipoarabinoman and APA/ModD antigen were differentially represented in both PPDs. This could explain the difference in stimulation induction of yIFN observed at individual level. Although PPDj-IB could not improve PPDa performance, we could easily produce an effective purified protein derivative for in vitro assays.

摘要

纯化蛋白衍生物(PPDs)是由分枝杆菌培养物制备的非限定抗原。它们通常用于评估动物和人类的特异性细胞免疫反应。牛型和禽型PPDs通常用作结核病和副结核病等分枝杆菌感染的抗原。然而,副结核分枝杆菌亚种的PPD(PPDj)既不常用也不常见。然而,鸟分枝杆菌亚种的PPD实际上是被使用的。我们旨在使用γ干扰素释放试验从本地分离的副结核分枝杆菌中获得并评估PPDj的性能。当使用这种名为PPDj-IB的抗原时,感染牛和对照牛之间γ干扰素释放的刺激存在显著差异。感染动物对两种抗原(PPDa和PPDj-IB)的刺激相似。然而,一些动物被PPDj-IB阳性刺激,而未被PPDa刺激。我们通过蛋白质印迹法证明,两种抗原分子,脂阿拉伯甘露聚糖和APA/ModD抗原在两种PPDs中的表达存在差异。这可以解释在个体水平上观察到的γ干扰素刺激诱导的差异。虽然PPDj-IB不能提高PPDa的性能,但我们可以很容易地生产出一种用于体外试验的有效的纯化蛋白衍生物。

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