• 미생물학회지
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• 제40권1호
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• pp.23-28
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• 2004

단순포진 바이러스 감염을 유발하는Herpes simplex2형 바이러스의 감염에 관여하는 항원들과 중화항체 생산을 유발하는 주요 항원들의 위치를 확인하였다. Vero cell에 감염하였을 때 48시간 동안 31, 43, 59, 69 kDa 바이러스 항원들이 지속적으로 발현되었으며, 감염된 쥐에서 생산한 항체와의 반응에서는 51 kDa 항원이 가장 강한 반응을 보였다. 면역전자현미경으로 위치를 확인한 결과 colloidal gold가 바이러스 표면에 발견되는 것으로 보아 이 항원이 바이러스 표면에 존재하고 있는 것으로 확인되었다. 형광현미경 분석은 이 항원들이 감염된 세포 내에서 전반적으로 발견되었고 특히 세포 표면에서 많이 발현되고 있었다.

• 한국어병학회지
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• 제12권2호
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• pp.89-99
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• 1999

V. vulnificus ATCC 27562균주의 배양 온도를 $2^{\circ}C $, 20분간 및 6% 에탄올, 10분간으로 반응시켰을 때 SDS-PAGE상에서 새로운 16가지의 heat shock protein(hsps)과 10가지의 ethanol shock protein이 나타났다. Lethal temperature에 노출하기전에 미리 열 충격을 가한 경우 thermo tolerance가 유도되었다. 균체면역에 의해 생성된 항혈청과 열 충격 세포에서 분리된 막단백질과의 ELISA에서는 Outer Membrane Protein(OMP)에서 높은 면역반응을 나타냈으며 western blotting으로는 Inner Membrane Protein(IMP)에서는 62kDa, OMP에서는 69 kDa단백이 높은 면역원성을 나타냈다. ethanol 충격 반응에서는 IMP에서는 48 kDa, OMP에서는 오직 major밴드에서만 면역반응성이 확인되었다. anti-V, vulnificus혈청에 대한 균체 응집시험에서는 열 충격 반응 후의 균체가 정상 균체에 비해 응집반응성이 높았다.

• BMB Reports
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• 제33권1호
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• pp.92-95
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• 2000

The HIV-1 p24(202-221) sequence ETINNEEEWDRVHPV HAGP contains a B-cell epitope with the earliest immune response and the highest antibody titer against anti-mouse sera obtained by immunization with p24 antigens. A novel mouse monoclonal antibody (mAb) was generated against the immunodominant B-cell epitope of the HIV-1 p24 capsid protein, p24(202-221). BALB/c mice were immunized with the four branched multiple antigenic peptide (MAP) containing the HIV-1p24(202-221) sequence, and antibody-secreting hybridoma were produced by fusion of mouse splenocytes with P3X63Ag8.653, mouse myeloma cells. One clone which produced the antigen-specific mAb named KI-24 (Isotype IgG1, light chain: ${\kappa}$) was identified. mAb KI-24 was highly specific for both the p24(202-221) and p24 proteins when analyzed by ELISA and Western blotting. Since p24(202-221) also contains a cytotoxic T-lymphocyte epitope, this specfic peptide epitope and the monoclonal antibody with specific reactivity against the p24 protein and p24(202-221) can be used in peptide vaccine development and p24 antigen detection from HIV patients.

• Parasites, Hosts and Diseases
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• 제43권1호
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• pp.19-25
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• 2005

Protective efficacy of vaccination with Neospora caninum multiple recombinant antigens against N. caninum infection was evaluated in vitro and in vivo. Two major immunodominant surface antigens (NcSAG1 and NcSRS2) and two dense granule proteins (NcDG1 and NcDG2) of N. caninum tachyzoites were expressed in E. coli, respectively. An in vitro neutralization assay using polyclonal antisera raised against each recombinant antigen showed inhibitory effects on the invasion of N. caninum tachyzoites into host cells. Separate groups of gerbils were immunized with the purified recombinant proteins singly or in combinations and animals were then challenged with N. caninum. Following these experimental challenges, the protective efficacy of each vaccination was determined by assessing animal survival rate. All experimental groups showed protective effects of different degrees against experimental infection. The highest protection efficacy was observed for combined vaccination with NcSRS2 and NcDG1. Our results indicate that combined vaccination with the N. caninum recombinant antigens, NcSRS2 and NcDG1, induces the highest protective effect against N. caninum infection in vitro and in vivo.

• 한국임상수의학회지
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• 제23권1호
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• pp.9-13
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• 2006

Anaplasma phagocytophilum의 주요 표면단백질인 Msp2 (p44)는 세균의 표면에 발현되는 주요한 항원 단백질이다. 본 실험에서는 A. phagocytophilum이 주요 숙주세포인 호중구나 HL-60 세포에 침입하는 데 있어, 세균의 주요 표면 단백질인 Msp2와 숙주세포의 표면에 발현되는 PSGL-1과의 반응 여부를 알아보았다. 그 결과, 재조합 단백질인 Msp2나 순수하게 분리된 A. phagocytophilum이 PSGL-1/FucT IV 유전자가 형질전환되어 PSGL-1이 발현되는 BJAB 세포와는 결합하지만, 순수한 BJAB 세포와는 전현 반응하지 않는 것을 관찰할 수 있었다(p<0.01 & p<0.01). 또한, 순수하게 분리된 A. phagocytophilum과 형질전환된(PSGL-1/FucT IV) BJAB 세포와의 결합이 Msp2의 단-클론항체나 Msp2 재조합 단백질의 농도에 따라 억제됨을 관찰할 수 있었다(p<0.05 & p<0.01). 따라서 A. phagocytophilum의 Msp2가 숙주세포인 호중구의 표면에 발현되는 PSGL-1과 직접적으로 결합하는 부착물질임을 알 수 있었다.

• Genomics & Informatics
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• 제20권1호
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• pp.11.1-11.20
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• 2022

Vibrio harveyi belongs to the Vibrio genus that causes vibriosis in marine and aquatic fish species through double-stranded DNA virus replication. In humans, around 12 Vibrio species can cause gastroenteritis (gastrointestinal illness). A large amount of virus particles can be found in the cytoplasm of infected cells, which may cause death. Despite these devastating complications, there is still no cure or vaccine for the virus. As a result, we used an immunoinformatics approach to develop a multi-epitope vaccine against most pathogenic hemolysin gene of V. harveyi. The immunodominant T- and B-cell epitopes were identified using the hemolysin protein. We developed a vaccine employing three possible epitopes: cytotoxic T-lymphocytes, helper T-lymphocytes, and linear B-lymphocyte epitopes, after thorough testing. The vaccine was developed to be antigenic, immunogenic, and non-allergenic, as well as having a better solubility. Molecular dynamics simulation revealed significant structural stiffness and binding stability. In addition, the immunological simulation generated by computer revealed that the vaccination might elicit immune reactions in the actual life after injection. Finally, using Escherichia coli K12 as a model, codon optimization yielded ideal GC content and a higher codon adaptation index value, which was then included in the cloning vector pET2+ (a). Altogether, our experiment implies that the proposed peptide vaccine might be a good option for vibriosis prophylaxis.

• Parasites, Hosts and Diseases
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• 제30권3호
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• pp.201-208
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• 1992

Theileria sergenti merozoite의 면역원성 polypeptide중 29, 34, 35 그리고 105KD를 함유한, 수용성 항원을 흘스타인 송아지 (1개월령)에게 접종한 후, 제주도의 야외 목장에 방목시킴으로써 진드기로부터 병원성 충주의 도전에 따른 항원의 효력, 안전성 그리고 방어력을 관찰하였다. 즉 송아지 20두에 수용성 항원(100 mg/dose)을 Freund's adjuvant와 함께 1차 접종, 4주 후에 추가접종하였으며, 다른 20두의 송아지를 대조군으로 사용하였다. 추가접종 5주 후에 이들 송아지를 야외 목장에 방목, 매월 혈액학적 그리고 생화학적 변화를 관찰하였다 예방접종군에 있어서 hematocrit와 총적혈구수는 진드기 도전 전과 비교하여 약간의 변화를 인정할 수 있었다. 일반적인 혈액학적 소견은 예방접종군과 비교하여 유의한 차이를 인정할 수 있었다(p<0.05). 7월에 있어서 적혈구내 기생률은 예방접종군에 있어서는 0.4%이었으나, 대조군에 있어서는 약 3.6%이었다. 체중 증가율, albumin, aspartate alninotransferase, total protein, 그리고 bilirubin에 있어서도 유의한 차이 (p<0.05)를 인정할 수 있었다. 예방접종군의 혈액학적 그리고 생화학적 측정치는 대조군 보다는 정상치에 접근하였다. 실험종료 직후에 있어서 예방접종군의 30% 송아지와 대조군의 모든 송아지의 치료와 대조군 중 25%는 수혈이 요구되었다. T. sergenti merozoite의 수용성 항원에 대한 이 같은 연구 결과는 앞으로 유전공학적 백신제조를 위한 연구자료로 활용될 수 있을 것이다.

• Parasites, Hosts and Diseases
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• 제53권4호
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• pp.403-411
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• 2015

Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes. Although many P. vivax proteins have been discovered, their functions are largely unknown. Among them, P. vivax reticulocyte binding proteins (PvRBP1 and PvRBP2) recognize and bind to reticulocytes. Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes. PvRBP1 and PvRBP2 are large (>326 kDa), which hinders identification of the functional domains. In this study, the complete genome information of the P. vivax RBP family was thoroughly analyzed using a prediction server with bioinformatics data to predict B-cell epitope domains. Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray. The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa. Evaluation of the humoral immune response to each recombinant PvRBP protein indicated a high antigenicity, with 38-88% sensitivity and 100% specificity. Of them, N-terminal parts of PvRBP2c (PVX_090325-1) and PvRBP2 like partial A (PVX_090330-1) elicited high antigenicity. In addition, the PvRBP2-like homologue B (PVX_116930) fragment was newly identified as high antigenicity and may be exploited as a potential antigenic candidate among the PvRBP family. The functional activity of the PvRBP family on merozoite invasion remains unknown.

• 대한수의학회지
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• 제45권1호
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• pp.45-53
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• 2005

GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

• 한국육종학회지
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• 제43권2호
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• pp.115-119
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• 2011

Soybean proteins are widely used for human and animal feeds worldwide. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are present in the raw mature soybean. P34 protein, referred as Gly m Bd 30K, has been identified as a predominant immunodominant allergen. The objective of this research is to identify the genetic mode of P34 protein for the improvement of soybean cultivar with a very low level of P34 protein. Two $F_2$ populations were developed from the cross of "Pungsannamulkong" ${\times}$ PI567476 and "Gaechuck2ho" ${\times}$ PI567476 (very low level of P34 protein). Relative amount of P34 protein was observed by Western blot analysis. The observed data for the progeny of "Pungsannamulkong" and PI567476 were 133 seeds with normal content of P34 protein and 35 seeds with very low level of P34 protein (${\chi}^2=1.157$, P=0.20-0.30). For the progeny of "Gaechuck#1" and PI567476, the observed data were 177 seeds with normal content of P34 protein and 73 seeds with very low level of P34 protein (${\chi}^2=2.353$, P=0.10-0.20). From pooled data, observed data were 310 seeds with normal content of P34 protein and 108 seeds with very low level of P34 protein (${\chi}^2=0.156$, P=0.50-0.70). The segregation ratio (3:1) and the Chi-square value obtained from the two populations suggested that P34 protein in mature soybean seed is controlled by a single major gene. Single gene inheritance of P34 protein was confirmed in 32 $F_2$ derived lines in $F_3$ seeds, which were germinated from the low level of P34 protein obtained from the cross of "Pungsannamulkong" and PI567476. These results may provide valuable information to breed for new soybean line with low level of P34 protein and identification of molecular markers linked to P34 locus.