• 대한바이러스학회지
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• 제29권2호
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• pp.107-118
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• 1999

To characterize the molecular nature of human immunodeficiency virus (HIV)-1, we determined the full-length HIV-1 sequences from cultured peripheral blood mononuclear cells (PBMC) of a Korean long-term nonprogressor (LTNP). Without antiretroviral therapy, the individual has maintained CD4+ T counts over $500/{\mu}l$ from 1989 to 1999. Plasma viral RNA copy was 992 U/ml in 1998. Culture supernatant showed positive from culture days 9. A series of 9 overlapping PCR products were amplified from cultured PBMC and cloned About 9.2 kb from R of 5' LTR to R of 3' LTR was determined by automated sequencing. The G-to-A hypermutations were shown throughout the entire region. As a result of G to A hypermutations, premature stop codon was found in integrase coding region. Though there was no recombination between subtypes over all genomes, TATA box in both LTRs was TAAAA which is detected in subtype E instead of TATAA in subtype B. And, there were nucleotide GC insertion between $NF-{\kappa}B$ I and Sp1 III, and duplication of $TCF-1{\alpha}$ in LTR. We could not find any deletion of amino acid in Nef, Gag, Pol and Env gene. This study is the first report on molecular nature of full genomes of HIV-1 isolated in Korea.

• 대한바이러스학회지
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• 제30권3호
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• pp.161-170
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• 2000

Continuous efforts are being made to find effective therapeutic agents against HIV-1, the causative agents of AIDS. In this study, we developed a cell-based assay system employing a trans-complementation for production of recombinant viruses which are capable of undergoing one round of replication in CD4+ T cells. This assay system was tested for ability to screen the agents that act at late stage of HIV-1 life cycle. The effect of a protease inhibitor on the trans-complementation assay was assessed. Recombinant HIV-1 viruses were prepared from a trans-complementation in the presence of various concentrations of protease inhibitor. Inhibition of single round infection of these recombinant viruses by protease inhibitor was observed to be a dose-dependent manner. Inhibitory effects of a protease inhibitor on HIV-1 Gag polyprotein processing by HIV-1 protease was detected at concentrations of the protease inhibitor compatible with inhibition of virus infection, confirming that the corresponding step was involved in the inhibitory mechanism of this compound. Together, these results provide evidence that a cell-based assay system established in this study can be used to screen the agents that target the late stage of HIV-1 life cycle.

• Journal of Veterinary Science
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• 제24권1호
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• pp.15.1-15.13
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• 2023

Background: Inactivated vaccines are limited in preventing foot-and-mouth disease (FMD) due to safety problems. Recombinant virus-like particles (VLPs) are an excellent candidate for a novel vaccine for preventing FMD, given that VLPs have similar immunogenicity as natural viruses and are replication- and infection-incompetent. Objectives: The 3C protease and P1 polyprotein of type O FMD virus (FDMV) was expressed in yeast Hansenula polymorpha to generate self-resembling VLPs, and the potential of recombinant VLPs as an FMD vaccine was evaluated. Methods: BALB/c mice were immunized with recombinant purified VLPs using CpG oligodeoxynucleotide and aluminum hydroxide gel as an adjuvant. Cytokines and lymphocytes from serum and spleen were analyzed by enzyme-linked immunosorbent assay, enzyme-linked immunospot assay, and flow cytometry. Results: The VLPs of FMD were purified successfully from yeast protein with a diameter of approximately 25 nm. The immunization of mice showed that animals produced high levels of FMDV antibodies and a higher level of antibodies for a longer time. In addition, higher levels of interferon-γ and CD4⁺ T cells were observed in mice immunized with VLPs. Conclusions: The expression of VLPs of FMD in H. polymorpha provides a novel strategy for the generation of the FMDV vaccine.

• IMMUNE NETWORK
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• 제23권2호
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• pp.16.1-16.19
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• 2023

Bacillus Calmette-Guerin (BCG) vaccine is the only licensed vaccine for tuberculosis (TB) prevention. Previously, our group demonstrated the vaccine potential of Rv0351 and Rv3628 against Mycobacterium tuberculosis (Mtb) infection by directing Th1-biased CD4⁺ T cells co-expressing IFN-γ, TNF-α, and IL-2 in the lungs. Here, we assessed immunogenicity and vaccine potential of the combined Ags (Rv0351/Rv3628) formulated in different adjuvants as subunit booster in BCG-primed mice against hypervirulent clinical Mtb strain K (Mtb K). Compared to BCG-only or subunit-only vaccine, BCG prime and subunit boost regimen exhibited significantly enhanced Th1 response. Next, we evaluated the immunogenicity to the combined Ags when formulated with four different types of monophosphoryl lipid A (MPL)-based adjuvants: 1) dimethyldioctadecylammonium bromide (DDA), MPL, and trehalose dicorynomycolate (TDM) in liposome form (DMT), 2) MPL and Poly I:C in liposome form (MP), 3) MPL, Poly I:C, and QS21 in liposome form (MPQ), and 4) MPL and Poly I:C in squalene emulsion form (MPS). MPQ and MPS displayed greater adjuvancity in Th1 induction than DMT or MP did. Especially, BCG prime and subunit-MPS boost regimen significantly reduced the bacterial loads and pulmonary inflammation against Mtb K infection when compared to BCG-only vaccine at a chronic stage of TB disease. Collectively, our findings highlighted the importance of adjuvant components and formulation to induce the enhanced protection with an optimal Th1 response.

• 생물정신의학
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• 제1권1호
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• pp.98-108
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• 1994

The etiology and pathophysiology of schizophrenia remain unknown. It has been postulated that infectious-autoimmune process may play a role in the pathogenesis of symptoms in some schizophrenic patients. Findings of altered interleukin(IL) regulation have been regarded as additional proof that schzophrenia has an infectious-autoimmune background. In the present study, we measured mitogen-stimulated production of and serum level of IL-$1{\beta}$, IL-2, IL-6 using ELISA in 16 neuroleptic-free schizophrenic patients and in 16 age, sex matched healthy controls. The results were as follows : 1) There was a significant decrease of IL-2 production in schizophrenic patients than in normal controls(respectively $1.90{\pm}0.13ng/m{\ell}$, $2.79{\pm}0.14ng/m{\ell}$, p<0.001). But there was no significant difference of IL-$1{\beta}$ production and IL-6 production between schizophrenic patients and normal controls. 2) There was a significant increase of serum level of IL-2 in schizophrenic pateitns than in normal controls(respectively $184.8{\pm}12.8pg/m{\ell}$, $104.2{\pm}34.2pg/m{\ell}$, p<0.01). Serum level of IL-$1{\beta}$ was partially detected in both groups and serum level of IL-6 was not detected in both groups. 3) There was no significant differences of IL-$1{\beta}$, -2, -6 production & serum level of IL-2 according to male vs female, paranoid type vs undifferentiated type, drug-naive group vs drug-free group in schizophrenic patients. 4) There was significant correlation between IL-$1{\beta}$ and IL-6 production(r=0.86, p<0.001). No correlation between IL-$1{\beta}$, -2, -6 production, serum level of IL-2 and age, duration of illness, and BPRS score was found. It has been suggested that the low lymphocyte production of IL-2 in the patients with autoimmune disease occurs because the T cells are activated and lymphocyte-derived IL-2 has been released into the serum. The authors suggest that decreased IL-2 production in our schizophrenic patients is due to increased IL-2 serum level in those patients. Thus our finding of low IL-2 production and high serum level of IL-2 in our schizophrenic patients is compatible with the possibility that our patients have an autoimmune process. Further study on relationship between IL alteration and other immunological abnormalities(the presence of serum autoantibody and of anti-brain antibody, $CD4^+$, $CD8^+$ cell index, etc) in schizophrenic patients will be warranted.

• Experimental and Molecular Medicine
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• 제50권12호
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• pp.12.1-12.14
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• 2018

Osteoporosis develops with high prevalence in both postmenopausal women and hypogonadal men. Osteoporosis results in significant morbidity, but no cure has been established. Mesenchymal stem cells (MSCs) critically contribute to bone homeostasis and possess potent immunomodulatory/anti-inflammatory capability. Here, we investigated the therapeutic efficacy of using an infusion of MSCs to treat sex hormone-deficient bone loss and its underlying mechanisms. In particular, we compared the impacts of MSC cytotherapy in the two genders with the aim of examining potential gender differences. Using the gonadectomy (GNX) model, we confirmed that the osteoporotic phenotypes were substantially consistent between female and male mice. Importantly, systemic MSC transplantation (MSCT) not only rescued trabecular bone loss in GNX mice but also restored cortical bone mass and bone quality. Unexpectedly, no differences were detected between the genders. Furthermore, MSCT demonstrated an equal efficiency in rectifying the bone remodeling balance in both genders of GNX animals, as proven by the comparable recovery of bone formation and parallel normalization of bone resorption. Mechanistically, using green fluorescent protein (GFP)-based cell-tracing, we demonstrated rapid engraftment but poor inhabitation of donor MSCs in the GNX recipient bone marrow of each gender. Alternatively, MSCT uniformly reduced the $CD3^+T$-cell population and suppressed the serum levels of inflammatory cytokines in reversing female and male GNX osteoporosis, which was attributed to the ability of the MSC to induce T-cell apoptosis. Immunosuppression in the microenvironment eventually led to functional recovery of endogenous MSCs, which resulted in restored osteogenesis and normalized behavior to modulate osteoclastogenesis. Collectively, these data revealed recipient sexually monomorphic responses to MSC therapy in gonadal steroid deficiency-induced osteoporosis via immunosuppression/anti-inflammation and resident stem cell recovery.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2000년도 International Symposium on Bioinformatics
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• pp.21-22
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• 2000

Expressed sequence tags (EFTs) are the partial segments of cDNA produced from 5 or 3 single-pass sequencing of cDNA clones, error-prone and generated in highly redundant sets. Advancement and expansion of Genomics made biologists to generate huge amount of ESTs from variety of organisms-human, microorganisms as well as plants, and the cumulated number of ESTs is over 5.3 million, As the EST data being accumulate more rapidly, it becomes bigger that the needs of the EST analysis tools for extraction of biological meaning from EST data. Among the several needs of EST analyses, the extraction of protein sequence or functional motifs from ESTs are important for the identification of their function in vivo. To accomplish that purpose the precise and accurate identification of the region where the coding sequences (CDSs) is a crucial problem to solve primarily, and it will be helpful to extract and detect of genuine CD5s and protein motifs from EST collections. Although several public tools are available for EST analysis, there is not any one to accomplish the object. Furthermore, they are not targeted to the plant ESTs but human or microorganism. Thus, to correspond the urgent needs of collaborators deals with plant ESTs and to establish the analysis system to be used as general-purpose public software we constructed the pipelined-EST analysis system by integration of public software components. The software we used are as follows - Phred/Cross-match for the quality control and vector screening, NCBI Blast for the similarity searching, ICATools for the EST clustering, Phrap for EST contig assembly, and BLOCKS/Prosite for protein motif searching. The sample data set used for the construction and verification of this system was 1,386 ESTs from human intrathymic T-cells that verified using UniGene and Nr database of NCBI. The approach for the extraction of CDSs from sample data set was carried out by comparison between sample data and protein sequences/motif database, determining matched protein sequences/motifs that agree with our defined parameters, and extracting the regions that shows similarities. In recent future, in addition to these components, it is supposed to be also integrated into our system and served that the software for the peptide mass spectrometry fingerprint analysis, one of the proteomics fields. This pipelined-EST analysis system will extend our knowledge on the plant ESTs and proteins by identification of unknown-genes.

• 한국발생생물학회지:발생과생식
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• 제15권4호
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• pp.373-383
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• 2011

성 호르몬과 유사한 기능을 하는 것으로 알려진 내분비교란물질은 흰쥐 흉선세포에 세포자연사를 일으켜 흉선의 기능을 감소시키는 것으로 보고되고 있으나, 그 작용 기전에 대해서는 잘 알려져 있지 않다. 따라서 본 연구에서는 내분비 교란물질 중 하나인 Tributyltin(TBT)를 흰쥐에 투여한 후 흉선의 상피세포가 지방세포로 분화되는지를 조사하고, 이로 인한 T 세포의 세포자연사와 연관성을 조사함으로써 TBT가 흉선기능에 미치는 영향을 알아보고자 하였다. 3주령 된 암컷 흰쥐에 각각 TBT 1, 10, 25 mg/kg/day를 1주일 동안 경구 투여한 후, 흉선을 획득하여 무게와 세포 수를 측정하였고, 파라핀 절편으로 H&E 염색, TUNEL 분석을 수행하였다. 또한 real-time PCR 방법으로 상피세포 표지 유전자들, 지방세포유도 유전자들과 세포자연사 관련 유전자들을 비교 분석하였으며, 유세포 분석기를 통해 세포자연사 정도를 조사하였다. 흉선의 무게와 세포 수는 대조군과 비교하여 TBT의 농도가 높을수록 유의하게 감소하였다. H&E 염색 결과, TBT의 농도가 증가할수록 흉선의 크기가 감소하고 피질과 수질의 경계가 흐려짐을 관찰하였다. TUNEL 염색 결과에서는 대조군에 비해 TBT를 투여한 군에서 세포자연사가 증가하였으며, 유세포 분석결과에서도 TBT 농도 의존적으로 세포자연사가 증가하였다. Real-time PCR 결과, 상피세포 표지 유전자인 EVA, IL-7, AIRE, KGF는 TBT의 농도가 증가할수록 발현양이 유의하게 감소한 반면, 지방세포 유도와 관련된 유전자인 $PPAR{\gamma}$, aP2, PEPCK, CD36은 TBT의 농도가 증가할수록 발현양이 유의하게 증가하였다. 세포자연사와 관련된 유전자인 $TNF{\alpha}$, TNFR1도 TBT의 농도가 증가할수록 발현양이 유의하게 증가하였다. 위의 결과를 종합하여 볼 때, 내분비교란물질인 TBT는 흉선 상피세포에 직접적으로 작용하여 지방세포로 분화 유도시킴으로써 흉선의 기능을 상실케 만들면서 T 세포의 발달에 영향을 미치는 것으로 보인다. 이러한 결과는 지속적으로 노출되는 TBT가 흉선의 기능을 떨어뜨림으로써 면역력 저하를 유발시킬 수 있음을 제시하고 있다.

• Tuberculosis and Respiratory Diseases
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• 제46권1호
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• pp.44-52
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• 1999

연구배경: 기관지천식은 호산구성 기도내 염증반응을 특징으로 하는데 호산구의 침윤에 관여하는 cytokine으로 T 림프구에서 분비되는 IL-3, IL-5, GM-CSF 등이 잘 알려져 있다. 한편 IL-10은 항염증성 cytokine으로 이러한 cytokine 분비를 억제할뿐만아니라, 호산구에 대해서는 직접적으로 자사(apoptosis)를 유도하고 조직내 첨착을 억제하는 기능을 갖고 있다. 본 연구에서는 기관지천식 환자에서의 기관지폐포세척액과 말초 혈액단핵세포에서 분비되는 IL-10을 정상대조군과 비교하고 기도내 염증반응정도와의 연관성을 분석하여 기관지천식에서의 IL-10 역할을 알아보고자 하였다. 방 법: 기관지천식환자 23명, 정상대조군 11명을 대상으로 기관지폐포세척액내와 말초혈액단핵구에서 분비되는 IL-10을 ELISA 방법으로 측정하고, 기도의 염증반응정도는 기관지세포세척액내 총세포수 및 호산구분율과 기관지 생검조직내 호산구 침윤정도, methacholine 기관지유발 검사에서 $PC_{20}$값으로 평가하였다. 결 과: 기관지폐포세척액내 IL-10과 말초혈액단핵구에서 분비되는 IL-10은 기관지천식 환자군과 정상대조군 사이에 통계적으로 유의한 차이는 없었다. 기관지천식환자군에서 기관지폐포세척액내 IL-10은 기관지폐포세척액내 호산구분율 및 기관지 조직내 침윤된 호산구수와는 유의한 역상관관계를 보였고, 메타콜린에 대한 PC20에 따른 차이는 없었다. 말초혈액단핵구에서 분비되는 IL-10은 기관지폐포세척액내 IL-10과 상관 관계가 없었으며, 말초혈액내 호산구수나 eosinophilic cationic protein과도 유의한 상관관계가 없었다. 결 론: 기관지천식 환자군의 기관지폐포세척액내 IL-10은 기관지폐포세척액이나 기관지 조직내 호산구 침윤과 역상관관계를 보였지만, 정상대조군과 비교해서 IL-10 감소가 관찰되지 않았기 때문에 기관지천식에서는 항염증 효과가 있는 IL-10이 중요하게 작용하지 않을 것으로 생각된다.