초록 ▼

한국형 삼일열말라리아 퇴치를 위한 원충 및 매개모기의 생물전략적 실용화 연구를 위하여 삼일열말라리아의 인체숙주와 곤충숙주사이의 생활환을 차단하는 백신물질 (전파단백 Pv25, Pv28)의 특징을 규명 실용화를 위한 안전성 시험을 수행하였다.
삼일열말라리아 단백분해효소의 생물학적 특성을 구명하고 저해제를 응용하여 말라리아 치료제 개발의 가능성을 확립하였으며 표면항원 단백질의 면역학적 특성 분석과 이를 이용한 DNA 백신 개발을 시도하였고, 말라리아 항원에 대한 면역반응의 증강을 유도할 수 있는 기초연구자료를 수립하였다.
또

한국형 삼일열말라리아 퇴치를 위한 원충 및 매개모기의 생물전략적 실용화 연구를 위하여 삼일열말라리아의 인체숙주와 곤충숙주사이의 생활환을 차단하는 백신물질 (전파단백 Pv25, Pv28)의 특징을 규명 실용화를 위한 안전성 시험을 수행하였다.
삼일열말라리아 단백분해효소의 생물학적 특성을 구명하고 저해제를 응용하여 말라리아 치료제 개발의 가능성을 확립하였으며 표면항원 단백질의 면역학적 특성 분석과 이를 이용한 DNA 백신 개발을 시도하였고, 말라리아 항원에 대한 면역반응의 증강을 유도할 수 있는 기초연구자료를 수립하였다.
또한 한국인의 항 말라리아 제제 감수성 분석 및 내성균 추적을 위하여, 국내에서 사용 중인 항말라리아 제재의 작용 단계 및 기전 규명하였다.
한편, Anopheles속 모기의 매개능 연구를 위하여 말라리아 감염과 연관된 유전형을 분석 하고 유전자 표지의 개발 및 유전자발현 차이 규명하였고 말라리아 매개모기에 관한 계통학적, 생물학적 특성을 규명하기 위한 기초자료를 확보하였다. 7과제의 연계된 전략적 연구를 통하여 말라리아 퇴치를 위한 백신개발에 중요한 연구기반이 확립되었다.

Abstract ▼

To evaluate the effectiveness of transmission blocking vaccine, the laboratory cultivation of Anophelis mosquitoes, cloning of Pv25, Pv28 were conducted. It was found that Pv25, Pv28 can enhance good T-cell immunity only with small amount. The characterization was done with E.coli expression system

To evaluate the effectiveness of transmission blocking vaccine, the laboratory cultivation of Anophelis mosquitoes, cloning of Pv25, Pv28 were conducted. It was found that Pv25, Pv28 can enhance good T-cell immunity only with small amount. The characterization was done with E.coli expression system and simple toxixicity test was conducted with purified protein expressed by baculovirus expression system.
To verify the change of anti-malarial drug susceptibility, we analyzed the pharmakokinetic characteristics of chloroquine of prophylactic dose in Korean healthy individuals and therapeutic dose in Korean healthy individuals. We found that the resistance against prophylactic dose had already appealed since 2000, but the patients with the resistance was treated with chloroquine of therapeutic dose. The resistance against therapeuic dose was not found. Chloroquine eradicated efficiently Plasmodium of during schizogony, whereas gametocyte seemed to be refractory to chloroquine.
Mass rearing system of An. sinensis. An. pullus, An. lesteri which are most dominat species of Korean anopheline mosquitoes was established. Karyotypes, hybrid test and DNA sequence comparison for these species were performed. From susceptibility test, it is clearly verified that An. sinensis is not vivax malaria vector mosquito. Mass rearing system can be applied for insecide development, mosquito ethology, and susceptibility test.
In the study of immunology on malaria, stimulation of NKT cells enhanced cytotoxic activity against weak antigens such as B16 tumor antigens was found. Stimulation of NKT cell-associated innate immunity by using ${\alpha}$GalCer at the time of immunization with P. falciparum antigen cocktail greatly enhanced the generation of antigen-specific CD8+ T cell population and CTL responses against malarial antigens. MSA peptide cocktail antigen loaded Bone marrow-derived dendritic cells elicited expansion of the antigen-specific IFN-g secreting CD8+ T cells and induced antigen-specific CTL responses. Dendritic cells are potent antigen-presenting cells that play a crucial role in inducing malarial antigen-specific immune responses.
A novel proteases from p. vivax was identified. An analyses of genetic variations of drug-resistance genes and proteases genes in P. vivax wild isolates, an evaluation of cysteine protease inhibitors as antimalarial chemotherapeutics, and characterizations of biochemical and functional properties of cysteine and aspartic proteases of Plasmodium vivax were conducted.
Four major vaccine candidate proteins Circumsporozoite protein (CSP), Duffy-binding protein (DBP), merozoite surface protein-1 (MSP-1), apical membrane antigen-1 (AMA-1), were cloned into pcDNA 3.1(-) vector and expression plasmids were finally constructed. This study found that pAMA1+1L-12 vaccination enhance IFN-r production and may help to protect against the infection of P. vivax. These DNA plasmids are expected to be potential vaccines.
On the other hand , the study of gene expressions among the mosquitoes that blood-fed and sugar-fed can be very important because it will provide a broadened basis for understanding vector-parasite interactions. PCR amplification using species-specific primer identified Anopheles sinensis. The eggs from an An. sinensis female were reared in a separate pan in insectaries, F1 progenies of adults and larvae were used for molecular analysis. To determine the gene expression patterns, differentially expressed gene (DEG) were screened by ACP-based PCR using the Gene Fishing DEG kits. In blood-fed mosquitoes, total 68 DEGs were expressed at least three-fold above their levels in the sugar-fed mosquitoes and larvae. This result will provide a resource for improving annotation of the An. sinensis genome, Importantly, increased understanding of An. sinensis biology at the molecular level may open new avenues for intervention against malaria transmission regulation.

목차 Contents

• II. 총괄과제 연구결과...42
• 1. 연구과제의 배경 및 필요성...43
• 2. 국내외기술현황...45
• 3. 연구개발과제의 추진 체계...48
• 4. 연구 개발 수행 내용 및 결과...49
• 5. 목표달성도 및 관련분야 기여도...57
• 6. 향후 연구성과 추진계획...60
• III. 세부과제 연구결과...62
• 제 1 세부과제...62
• 제 2 세부과제...92
• 제 3 세부과제...107
• 제 4 세부과제...121
• 제 5 세부과제...134
• 제 6 세부과제...159
• 제 7 세부과제...181
• IV. 첨부서류...191