Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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The Purification and Immunogenicity of Pneumococcal Surface Protein (PspA) from Invasive Streptococcus pneumoniae KNIH1156 Isolated in Korea

국내 임상 분리주 Streptococcus pneumoniae KNIH1156으로부터 PspA 단백 항원의 정제 및 면역원성 확인

  • 정경석 (안산 1 대학 임상병리과) ;
  • 배송미 (국립보건원 세균부 호흡기 세균과)
  • Published : 2002.03.01
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Abstract

Pneumococcal surfacce protein A (PspA) is an important virulence factor and an antigenically variable surface protein of the pneumococci. To purify the PspA from S. pneumoniae KNIH1156 , a clinical isolate (type 19F), we have taken advantage of the fact that PspA is released from the surface of pneumococci into the medium by growing in a CDM-ET medium and PspA is capable of binding human lactoferrin, the iron carrier protein. PspA of S. pneumoniae KNIH1156 was purified from culture supernatant by human lactoferrin (hLf) affinity chromatography. The purified PspA was confirmed with anti-PspA antiserum and also had the binding capacity to hLf specifically. To determine whether the purified PspA could elicit protection in mice against pneumococcal inflection, we immunized the mice with purified PspA and subsequently challenged with S. pneumoniae KNIH1156. Immunization with purified PspA protected mice from 500 times the $LD^{50}$ of S. pneumoniae KNIH1156. Therefore, it has been shown that purified PspA fromS. pneumoniae KNIH1156 (type 19F) is a protective immunogen.

국내 분리 침습성 균주중선별된 S. pneumoniae KNIH1156 (type 19F)으로부터 페렴구균의 병원성 인자이며 항원학적으로 다양한 표면단백항원인 pneumococcal surface protein A (PspA)를 분리${\cdot}$정제하였다. 폐렴구균을 CDM-ET배지에서 배양하게 되면 배지내로 PspA가 방출된다는 점과 PspA가 인간의 lactoferrin에 특이적으로 결합한다는 사실을 이용하여 CDM-ET 배지에서 S. pneumoniae KNIH1156 을 배양한 후 배양액을 농축하여 lactoferrinaffinity chromatography에 통과시켜 PspA를 분리, 정제하였다. 정제 후 anti-PspA antiserum으로 PspA를 확인하여 순수분리, 정제되었음을 확인하였으며 또한 인간의 lactoferrin과의 결합능력을 유지하고 있음을 확인하였다. 순수하게 분리하여 정제된 PspA의 면역원성을 확인하기 위하여 ICR mice에 욕강주사하였을 때 $LD_{50}$ 가 $1{\times}10^{7.5}$ CFU/ml께서 $1{\times}10^{10}$ CFU/ml로 약 500배 중가함을 관찰하였다. 따라서 본 실험에서S. pneumoniae KNIH1156 으로부터 분리${\cdot}$ 정제한 PspA가 면역원성과 방어능을 가지고 있음을 확인할 수 있었다.

Keywords

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