• 제목/요약/키워드: Capsular polysaccharide

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OPTIMIZATION OF CULTURE CONDITIONS FOR PRODUCTION OF PNEUMOCOCCAL CAPSULAR POLYSACCHARIDE TYPE I

  • Kim, S.N.;K.K. Min;Kim, S.H.;Park, I.H.;Lee, S.H.;S.N. Pyo;D.K. Rhee
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.186-186
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    • 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.

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Improvement of the Stability of the Multibead Assay for Pneumococci by the Use of 6B-protein Complex (피막 다당류 6B-단백질 복합체를 이용한 폐렴구균에 대한 Multibead Assay의 안정성 향상)

  • 김지혜;임낙룡;박문국
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.62-65
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    • 2003
  • A procedure to increase the stability of 6B capsular polysaccharide on microbead surface in the mutibead assay, a serotyping method for Streptococcus pneumoniae, was studied. Pneumococcal capsular polysaccharide 6B was conjugated to bvine serum albumin (BSA), and the coating efficiency and the stability of the 6B-BSA complex was measured. The 6B-BSA complex showed about 200-fold higher coating efficiency to polystyrene surface than 6B polysaccharide. And the stability of the 6B- BSA to be used in the multibead assay for 30 days after coating.

Characterization of Photobacterium damselae subsp. piscicida isolated from cultured starry flounder, Platichthys stellatus in Korea (우리나라 양식 강도다리, Platichthys stellatus에서 분리된 Photobacterium damselae subsp. piscicida의 특성)

  • Cho, Young Ah;Han, Hyun-Ja;Mun, Hee Eun;Jung, Sung Hee;Park, Myoung Ae;Kim, Jin Woo
    • Journal of fish pathology
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    • v.26 no.2
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    • pp.77-88
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    • 2013
  • Starry flounder, Platichthys stellatus (body length $4.4{\pm}0.51cm$) that became sick during an outbreak of disease at mariculture facilities at Ulsan, Korea in August of 2012, were examined to identify the cause of the disease. Diseased fish didn't show a unique sign, but the oxidase-positive and gram negative rod was isolated from moribund fish. The bacterium was revealed as Photobacterium damselae subsp. piscicida by biochemical analysis and sequence analysis of the 16S rRNA and capsular polysaccharide (CPS) genes. The isolates (AD5) was carrying susceptible to ofloxacin and gentamycin and showed high growth value at $18^{\circ}C$ and $25^{\circ}C$ compared to four other P. damsela strains.

Immunogenicity and Safety of Vi Capsular Polysaccharide Typhoid Vaccine in Healthy Persons in Korea

  • Lim, Sang-Min;Jung, Hahn-Sun;Kim, Min-Ja;Park, Dae-Won;Kim, Woo-Joo;Cheong, Hee-Jin;Park, Seung-Chul;Lee, Kwang-Chul;Shin, Young-Kyoo;Tan, Hyun-Kwang;Kim, Sang-Lin;Sohn, Jang-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.611-615
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    • 2007
  • The purpose of this study was to evaluate the immunogenicity and safety of Salmonella Typhi Vi capsular polysaccharide vaccine (Vi vaccine) in Korea. The immunogenicity of a single dose of Vi vaccine was evaluated in 157 subjects (75 children and 82 adults) before and at 1, 6, and 12 months after vaccination. Immunogenicity was measured with a passive hemagglutination assay (PHA), quantified as geometric mean titers (GMTs) and seroconversion rates. The safety of the vaccine was investigated by determining adverse reactions occurring within 4h, 3 days, and 1 month after injection. The seroconversion rate for children and adults 1 month after vaccination was 96.92% and 89.02%, respectively. In the case of children, the GMTs of Vi antibodies before vaccination were $5.87{\pm}1.34\;and\;142.59{\pm}2.39$ at one month after vaccination. For adults, the GMTs before and one month after vaccination were $5.58{\pm}1.28\;and\;58.56{\pm}3.67$, respectively. Vi antibodies persisted for as long as 6 and 12 months after vaccination. All adverse reactions in adults and children were minor and did not require treatment. The Vi CPS vaccine was safe and immunogenic in adults and children older than 5 years.

Overexpression of Capsular Polysaccharide Biosynthesis Protein in Lactobacillus plantarum P1 to Enhance Capsular Polysaccharide Production for Di-n-butyl Phthalate Adsorption

  • Liu, Wei-Bing;Lin, Zhi-Wei;Zhou, Ying;Ye, Bang-Ce
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1545-1551
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    • 2021
  • Exopolysaccharides (EPSs) such as capsular polysaccharide (CPS) are important bioactive carbohydrate compounds and are often used as bioenrichment agents and bioabsorbers to remove environmental pollutants like di-n-butyl phthalate (DBP). Among the EPS-producing bacteria, lactic acid bacteria (LAB) have gained the most attention. As generally recognized as safe (GRAS) microorganisms, LAB can produce EPSs having many different structures and no health risks. However, EPS production by LAB does not meet the needs of large-scale application on an industrial scale. Here, the capA gene (encoding CPS biosynthesis protein) was overexpressed in Lactobacillus plantarum P1 to improve the production of EPSs and further enhance the DBP adsorption capability. Compared with P1, the CPS production in capA overexpressed strain was increased by 11.3 mg/l, and the EPS thickness was increased from 0.0786 ± 0.0224 ㎛ in P1 to 0.1160 ± 0.0480 ㎛ in P1-capA. These increases caused the DBP adsorption ratio of P1-capA to be doubled. Overall, the findings in this study provide a safe method for the adsorption and removal of DBP.

Adjuvant Effect of Polysaccharides from Aboveground Parts of Astragalus membranaceus (황기 지상부 다당체의 면역 및 백신보조 효과)

  • Yang, Su Jin;Lee, Si Young;Lee, Hannah;Park, Yeong Chul;Choi, Seon Kang;Yu, Chang Yeon;Chung, Ill Min;Lim, Jung Dae
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.5
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    • pp.408-419
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    • 2016
  • Background: In recent years, adjuvants have received increasing attention owing to the development of purified subunit and synthetic vaccines which are poor immunogens and require additional adjuvants to evoke an immune response. Therefore, immunologic adjuvants have been developed and tested. Plant polysaccharides have been recognized as effective biological response modifiers with low toxicity. Methods and Results: In this study, the polysaccharide from the aboveground part of Astragalus membranaceus Bunge containing immunomodulating arabino-3,6-galactan was evaluated for its hemolytic activity and adjuvant potential in the specific cellular and humoral immune responses to ovalbumin. The polysaccharide from the aboveground part of Astragalus membranaceus Bunge was co-immunized with the purified Vi capsular polysaccharide of Salmonella typhi vaccine in mice. The polysaccharide from the aboveground part of Astragalus membranaceus Bunge did not induce any hemolytic activity or side effects at doses up to $500{\mu}g/m{\ell}$. The concanavalin A-, lipopolysaccharide-, and ovalbumin-induced splenocyte proliferation and serum ovalbumin-specific IgG, IgG1 and IgG2b antibody titers in immunized mice were significantly enhanced by AMA. Pharmacological data revealed that the polysaccharide from the aboveground part of Astragalus membranaceus Bunge increased antigen-specific antibody levels in immunized mice. The polysaccharide from the aboveground part of Astragalus membranaceus Bunge-adjuvanted purified Vi capsular polysaccharide of Salmonella typhi vaccine improved the proliferation of splenocytes and macrophages as well as stimulated cytokine production. Conclusions: These results suggest that the polysaccharide from the aboveground part of Astragalus membranaceus Bunge-adjuvanted vaccines enhanced humoral and cellular immunity and that the polysaccharide from the aboveground part of Astragalus membranaceus Bunge is a safe and efficacious adjuvant candidate suitable for use in prophylactic and therapeutic vaccines.

Optimization of Culture Conditions for Production of Pneumococcal Capsular Polysaccharide Type I

  • Kim, Su-Nam;Min, Kwan-Ki;Kim, Seung-Hwan;Choi, In-Hwa;Lee, Suhk-Hyung;Pyo, Suhk-Noung;Rhee, Dong-Kwon
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.179-183
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    • 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.

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Measurement of Free Polysaccharide in Tetanus Toxoid-Conjugate Vaccine Using Antibody/Ammonium Sulfate Precipitation

  • Yoo, Tae-Hyeon;Kim, Hyun-Sung;Park, Sung-Sik;Bang, Eun-Young;Oh, Yong-K.;Kim, Li-Seop;Kim, Hun;Hur, Byung-Ki;Ryu, Yeon-Woo;Kim, Jong-Su
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.469-472
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    • 2003
  • A method that effectively precipitates capsular polysaccharide of Haemophilus influenzae type b (polyribosylribitol phosphate, PRP) conjugated to tetanus toxoid (TT), PRP TT in a liquid vaccine has been developed to measure free PRP present in TT-conjugate vaccine. The method involves adding anti-TT antibody and ammonium sulfate to precipitate PRP-TT conjugate and measuring free PRP in tile supernatant. This new method provides a complete precipitation of the total PRP-TT, and provides an accurate and reproducible measurements of free PRP. The accuracy of the assay was confirmed by spiking known amounts of unconjugated PRP to PRP-TT conjugate, and the new method was found to have no effect on free PRP while precipitating PRP-TT. The published acid precipitation method did not produce reproducible results due to incomplete precipitation of PRP-TT, especially when the vaccine is formulated in a salt-buffered solution.

A Novel Complement Fixation Pathway Initiated by SIGN-R1 Interacting with C1q in Innate Immunity

  • Kang, Young-Sun
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.23-25
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    • 2008
  • Serum complement proteins comprise an important system that is responsible for several innate and adaptive immune defence mechanisms. There were three well described pathways known to lead to the generation of a C3 convertase, which catalyses the proteolysis of complement component C3, and leads to the formation of C3 opsonins (C3b, iC3b and C3d) that fix to bacteria. A pivotal step in the complement pathway is the assembly of a C3 convertase, which digests the C3 complement component to form microbial-binding C3 fragments recognized by leukocytes. The spleen clears microorganisms from the blood. Individuals lacking this organ are more susceptible to Streptococcus pneumoniae. Innate resistance to S. pneumoniae has previously been shown to involve complement components C3 and C4, however this resistance has only a partial requirement for mediators of these three pathways, such as immunoglobulin, factor B and mannose-binding lectin. Therefore it was likely that spleen and complement system provide resistance against blood-borne S. pneumoniae infection through unknown mechanism. To better understand the mechanisms involved, we studied Specific intracellular adhesion molecule-grabbing nonintegrin (SIGN)-R1. SIGN-R1, is a C-type lectin that is expressed at high levels by spleen marginal-zone macrophages and lymph-node macrophages. SIGN-R1 has previously been shown to be the main receptor for bacterial dextrans, as well as for the capsular pneumococcal polysaccharide (CPS) of S. pneumoniae. We examined the specific role of this receptor in the activation of complement. Using a monoclonal antibody that selectively downregulates SIGN-R1 expression in vivo, we show that in response to S. pneumoniae or CPS, SIGN-R1 mediates the immediate proteolysis of C3 and fixation of C3 opsonins to S. pneumoniae or to marginal-zone macrophages that had taken up CPS. These data indicate that SIGN-R1 is largely responsible for the rapid C3 convertase formation induced by S. pneumoniae in the spleen of mice. Also, we found that SIGN-R1 directly binds C1q and that C3 fixation by SIGN-R1 requires C1q and C4 but not factor B or immunoglobulin. Traditionally C3 convertase can be formed by the classical C1q- and immunoglobulin-dependent pathway, the alternative factor-B-dependent pathway and the soluble mannose-binding lectin pathway. Furthermore Conditional SIGN-R1 knockout mice developed deficits in C3 catabolism when given S. pneumoniae or its capsular polysaccharide intravenously. There were marked reductions in proteolysis of serum C3, deposition of C3 on organisms within SIGN-$R1^+$ spleen macrophages, and formation of C3 ligands. The transmembrane lectin SIGN-R1 therefore contributes to innate resistance by an unusual C3 activation pathway. We propose that in the SIGN-R1 mediated complement activation pathway, after binding to polysaccharide, SIGN-R1 captures C1q. SIGN-R1 can then, in association with several other complement proteins including C4, lead to the formation of a C3 convertase and fixation of C3. Therefore, this new pathway for C3 fixation by SIGN-R1, which is unusual as it is a classical C1q-dependent pathway that does not require immuno globulin, contributes to innate immune resistance to certain encapsulated microorganisms.

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Inhibition of Yeast Growth by Broadly Cross-Reactive Antisera Elicited by Heterologous Mannan-Protein Conjugate

  • Machova, Eva;Korcova, Jana;Cizova, Alzbeta;Bystricky, Slavomir
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1177-1179
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    • 2015
  • A new approach to obtain broadly cross-reactive antisera against important yeast pathogens by intensive hyperimmunization with polysaccharide-protein conjugates is described here. Surface mannan of Candida albicans and capsular galactoglucoxylomannan of Cryptococcus laurentii were isolated and chemically linked to human serum albumin. Antisera elicited by a 7-week vigorous immunization of rabbits with the conjugates showed effective cross-reactive growth inhibition of different representatives of Candida spp. as well as Cryptococcus spp. IgG antibodies are evidenced as the effective component of the antisera.