• 약학회지
• /
• 제48권6호
• /
• pp.345-351
• /
• 2004

ln the present work to determine effect of a Streptococcus pneumoniae conjugate vaccine, S.pneumoniae capsule attached to the surface protein (JY-Pol) was ex amined. This JY-Pol contained approximately 92% and 6% carbohydrate and protein, respectively. Gel electrophoresis revealed the presence of the surface protein in the JY-Pol. By the double immunodiffusion and isotyping ELISA analyses, administration of JY-Pol that was adsorbed to alum adjuvant (JY-Pol/Alum) into mice induced IgM, IgG, and IgA specific for the S.pneumoniae capsule. The ATCC capsular polysaccharide adsorbed to alum (ATCC-Pol/Alum) provoked only IgM in mice. In survival tests, mice that were immunized with the JY-Pol/Alum before intravenous challenge with live S.pneumoniae survived entire period of 46 day-observation, whereas all mice that received ATCC-Pol/Alum or only diluent instead of the vaccination died within 5 and 12 days, respectively. Results from footpad-edema test showed that JY-Pol/Alum formula provoked the cellular immunity as determined by swelling of the mouse footpad. These data indicate that the naturally conjugated JY- Pol enhances resistance of mice against disseminated pneumococcal disease due to S.pneumoniae by both humoral and cellular immune responses.

• 약학회지
• /
• 제48권6호
• /
• pp.369-373
• /
• 2004

We previously reported that Streptococcus pneumoniae capsule attached to the surface protein (JY-Pol) was protective to systemic pneumococcal infection. The JY -Pol antigen induced IgM, IgG, and IgA in mice and provoked cell-mediated immunity. In this current study, we investigated the effect of anti JY-Pol antiserun and monoclonal antibody C2 (Mab C2) specific for the JY-Pol antigen against the pneumococcal disease. Mice that were given the antiserum survived longer than mice that received antiserum pre-absorbed with S.pneumoniae cells or DPBS as a negative control. Heat-treated anti JY-Pol antiserum resulted in survival rates similar to intact fresh JY-Pol antiserum. Mab C2 isolated from JY-Pol-immunized mice also enhanced resistance of naive mice against the pneumococcal diseaser. This protection by Mab C2 appeared to be mediated by opsonization as determined in a RAW 264.7 monocyte/macrophage cell line. Epitope analysis showed that Mab C2 epitope consisted of glucuronic acid and glucose that blocked the interaction of JY-Pol to the C2. Taken together, these data indicate that the antiserum induced by the JY-Pol, a naturally pneumococcal conjugate formula, mediated the protection by passive transfer, which was confirmed by protective effect of Mab C2.

• Clinical and Experimental Pediatrics
• /
• 제48권11호
• /
• pp.1206-1211
• /
• 2005

목 적 : 현재 연구 개발되고 있는 PspA에 근거한 폐구균 단백 백신은 기존의 백신과 달리 여러 종류의 혈청형에 대해 동시에 광범위한 방어 면역을 유도할 가능성이 있다. 동물 실험에서 PspA는 폐구균 보균과 호흡기 감염 뿐 아니라 패혈증에 대해서도 방어 면역을 형성함이 보고되었다. 본 연구는 한국 소아와 어른에게서 분리된 폐구균에서 PspA 백신에 포함된 PspA family인 family 1과 family 2의 분포를 알기 위해 시행되었다. 방 법 : 총 89주의 폐구균을 대상으로 항혈청을 이용하여 슬라이드 응집 방법으로 폐구균 피막의 혈청형을 분류하였다. 또한 PCR을 이용하여 family 1과 family 2의 시발체를 사용하여 분리된 폐구균의 PspA family를 정하였다. 결 과 : 총 89주의 폐구균 분리주에서 17 종류의 폐구균 혈청형이 분류되었다. PspA 종류는 79주(88.8%)에서 결정되었는데 20주(22.5%)가 family 1, 59주(66.3%)가 family 2이었다. 아홉주(10.1%)에서 family 1과 2에 모두 양성 반응을 보였으므로 family 1과 2 단백을 포함하는 PspA 백신의 가능한 방어 범위는 98.9%이었다. 각 PspA family들은 균이 분리된 사람의 나이, 분리된 곳, 혹은 페니실린에 대한 감수성 등과 의미 있는 관련성이 없었으나 피막 혈청형에 따라서는 의미 있는 차이를 보였다. 결 론 : 본 연구에서 분리된 폐구균 분리주의 98.9%가 PspA family 1과 2에 속하였으므로 이를 포함한 백신은 폐구균에 대해 혈청형과 관련 없이 좀 더 광범위한 효과를 보일 것으로 추정된다. 향후 좀더 다양하고 많은 폐구균 분리주를 대상으로 PspA family의 연구가 이루어진다면 PspA 백신의 개발과 적용에 많은 도움이 될 것으로 생각된다.

• Archives of Pharmacal Research
• /
• 제19권3호
• /
• pp.173-177
• /
• 1996

The Pneumococcus, Streptococcus pneumoniae, has an ample polysaccharide (PS) capsule that is highly antigenic and is the main virulence factor of the organism. The capsular PS is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 4 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based medium were found to support luxuriant growth of pneumococcus type 4 at the same level. Therefore in this study, the Casitone based medium was used to study optimization of the culture condition because of BHI broth's high cost and complex nature. The phase of growth which accomodated maximum PS production was exponential phase. Concentrations of glucose greater than 0.8% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplementation of various concentrations of metal ion (with the exception of calcium, copper, and magnesium ions) had adverse effects on growth and PS production. On the other hand, low level aeration and supplementation of 3 mg/l concentration of asparagine, phenylalanine, or threonine were beneficial for increased PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1996년도 춘계학술대회
• /
• pp.186-186
• /
• 1996

Streptococcus pneumoniae (pneumococcus), the most common cause of bacterial pneumonia, has an ample polysaccharide(PS) capsule that is highly antigenic and is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 1 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based media were found to support luxuriant growth of pneumococcus type 1 at the same level. Because BHI medium is rather expensive and more complex than the Casitone based media, the Casitone based media was used to study optimization of the culture condition. The phase of growth which accomodated maximum PS production was logarithmic phase. Concentrations of glucose greater than 0.2% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplemention of various concentrations of metal ion (with the exception of calcium ion) had adverse effects on growth and PS production. On the other hand, low level aeration was beneficial for increased PS production. Addition of 3 mg/I concentration of methionine, phenylalanine, and threonine were found to enhance growth and PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

• Journal of Microbiology
• /
• 제34권2호
• /
• pp.179-183
• /
• 1996

Streptoccus Pneumoniae (pneumococcus), the most common cause of bacterial pneumonia, has an ample polysaccharide (PS) capsule that is highly antigenic and is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsulard PS by type 1 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based media were found to support luxuriant growth of pneumococcus type 1 at the same level. Because BHI medium is rather expensive and more complex than the Casitone based media, the Casitone based media was uwed to study optimization of the culture condition. The phase of growth which accomodated maximum PS production was logarithmic phase. Concentrations of glucose greater than 0.2% did not ehnahce growth or PS production. Substitution of netrogen sources with other resources or supplementation of various concentrations of metal ion (with the exception of calcium ion) had adverse affects on growth and PS production. On the other hand, low level aeration was beneficial for increased PS production. Addition of 3 mg/1 concentration of methionine, phenylalanine, and threonine were found to enhance growth and PS production. The synerigistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cummulative increase in capsular PS production.

• 약학회지
• /
• 제49권6호
• /
• pp.484-489
• /
• 2005

Streptococcus pmeumoniae is the leading cause of pneumonia and bacterial meningitis. The current polysaccharide vaccine has been reported ineffective in elderly adults and children less than 2 years of age. Thus, in recent many researchers have been focused on a different approach, DNA vaccine. In our laboratory we developed a Streptococcus pneumoniae DNA (SPDNA) vaccine. This SPDNA vaccine was formulated by inserting the region encoding part of the capsule in the S. pneumoniae into the LAMP-1. In present work, with use of the SPDNA vaccine we attempted to establish a certain methodology useful for evaluation of effectiveness and immunoresponse of a DNA vaccine. Results showed that the subcutaneous route was the most effective for production of antisera specific for S. pneumoniae in mice. By isotyping analyses, IgM, IgGl, IgG2a, and IgG2b were determined. In addition, INF-$\gamma$ and IL-4 were predominantly detected. Combination of those data resulted in a pattern of IgGl < IgG2a=IgG2b and INF$\gamma\>$ >IL-4, which indicates the inmmunity towards the Thl response predominantly; furthermore, the SPDNA vaccination induced resistance of the CD4+T lymphocyte-depleted mice against disseminated pneumococcal infection. These data appear to be possibly due to activation of CDS8+T cell-activation. Taken together, this methodology can be applied for evaluating efficacy and mode of action of a DNA vaccine as minimum critera.