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肺炎克雷伯菌35 kDa和36 kDa孔蛋白是不同血清型中的常见抗原,并可诱导调理抗体。

Klebsiella pneumoniae 35 and 36 kDa porins are common antigens in different serotypes and induce opsonizing antibodies.

作者信息

Alcántar-Curiel M D, García-Latorre E, Santos J I

机构信息

Laboratorio de Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Secretaría de Salud (SSa), México, D.F., Mexico.

出版信息

Arch Med Res. 2000 Jan-Feb;31(1):28-36. doi: 10.1016/s0188-4409(99)00083-1.

Abstract

BACKGROUND

Klebsiella pneumoniae is a major cause of neonatal sepsis and nosocomial infections in Mexico. Antibiotic therapy is the first choice for treatment but the increase in multiple resistance strains has forced scientists to look for alternative treatments, such as immunotherapy. In this work, we propose that porins could be a common antigen among four different capsular serotypes of Klebsiella pneumoniae for the production of immune sera with opsonizing capacity.

METHODS

The 35 and 36 kDa porins from four different serotypes of the bacteria were isolated by the Nikaido method followed by purification in Sephacryl column chromatography. The 36 kDa of serotype K8 was further purified by electroelution. The 35 and 36 kDa porins were used to obtain rabbit polyclonal antibodies (PolyAb) to the four serotypes and the 36 kDa from K8 for the production of monoclonal antibodies (MoAb). Antigenic reactivity of PolyAb and MoAb were analyzed by ELISA and WB and their opsonizing capacity for human PMN was measured by chemiluminescence (CL) using capsulated and non-capsulated bacteria.

RESULTS

Porins from the four strains showe electrophoretic homology and cross reaction by ELISA and WB. CL assays indicated that PolyAb opsonized heterologous strains and that MoAb perform this in the absence of capsule.

CONCLUSIONS

K.pneumoniae 35 and 36 kDa porins are common antigens for the four serotypes studied and induce opsonizing antibodies.

摘要

背景

肺炎克雷伯菌是墨西哥新生儿败血症和医院感染的主要病因。抗生素治疗是首选治疗方法,但多重耐药菌株的增加迫使科学家寻找替代治疗方法,如免疫疗法。在这项研究中,我们提出孔蛋白可能是四种不同荚膜血清型肺炎克雷伯菌的共同抗原,可用于生产具有调理能力的免疫血清。

方法

采用Nikaido方法从该细菌的四种不同血清型中分离出35 kDa和36 kDa的孔蛋白,随后在Sephacryl柱色谱中进行纯化。血清型K8的36 kDa孔蛋白通过电洗脱进一步纯化。使用35 kDa和36 kDa的孔蛋白获得针对四种血清型的兔多克隆抗体(PolyAb)以及针对K8血清型36 kDa孔蛋白的单克隆抗体(MoAb)。通过ELISA和WB分析PolyAb和MoAb的抗原反应性,并使用包膜和未包膜细菌通过化学发光(CL)测量它们对人中性粒细胞的调理能力。

结果

四种菌株的孔蛋白通过ELISA和WB显示出电泳同源性和交叉反应。CL分析表明,PolyAb可调理异源菌株,而MoAb在无荚膜的情况下也能发挥这种作用。

结论

肺炎克雷伯菌35 kDa和36 kDa的孔蛋白是所研究的四种血清型的共同抗原,并能诱导产生调理抗体。

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