• 제목/요약/키워드: human immunodeficiency virus type 1 (HIV-1)

검색결과 64건 처리시간 0.028초

진핵세포에서 HSV-1 Envelope 변이 단백질의 발현 및 발현 단백질의 특성 연구 (Expression and Characterization of the Human Immunodeficiency Virus Type 1 Mutant Envelope Glycoproteins in Mammalian Cells)

  • 류지윤;박진서
    • 대한바이러스학회지
    • /
    • 제29권3호
    • /
    • pp.183-193
    • /
    • 1999
  • Human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is synthesized as a 160 KDa precursor, gp160, that is cleaved by a cellular protease to form the gp120 and gp41 subunits. Mammalian expression vectors were designed that are capable of efficient expression of various mutant envelope glycoproteins derived from a molecular clone of HIV-1. To construct these vectors, one type of mutation was made at the gp120-gp41 cleavage site by oligonucleotide-directed mutagenesis. And another mutation was made to change amino acids in the membrane spanning region of HIV-1 gp41 important for membrane anchorage. Next, these two mutations were combined to generate a vector to have double mutations in cleavage site and membrane-spanning region. These mutants were transiently expressed in mammalian cells. The effect of these mutations on envelope glycoprotein synthesis, proteolytic processing and secretion was determined. In addition, cell surface expression and ability of the glycoprotein to induce syncytium formation were examined. This study provides a mammalian expression system that is capable of efficient expression and secretion of soluble gp160.

  • PDF

Expression of Human Immunodeficiency Virus Type 1 Tat Proteins in Escherichia coli and Application to Study Tat Functions

  • Park, Jin-Seu;Lee, Han-Gyu;Lee, Yoon;Kang, Young-Hee;Rhim, Hyang-Shuk;Choi, Soo-Young
    • BMB Reports
    • /
    • 제33권4호
    • /
    • pp.337-343
    • /
    • 2000
  • The human immunodeficiency virus type 1 (HIV-1), transactivator of transcription (Tat), is one of the viral gene products that is essential for HIV-1 replication. The HIV-l Tat protein regulates transcription from an HIV-1 long terminal repeat (LTR) and affects the gene expression of cellular proteins during infection. In order to develop an expression system to overexpress and simply purify HIV-1 Tat proteins, the HIV-1 Tat coding sequences that contain one or two exons were amplified using PCR and cloned into a pET vector, which contains a consecutive stretch of six histidine residues at the amino-terminus. The reconstituted vectors were overexpressed in the E. coli strain and the soluble recombinant proteins were purified to be homogeneity in a single step by $Ni^{+2}-nitrilotriacetic$ acid Sepharose chromatography under nondenaturing conditions. Recombinant HIV-1 Tat proteins were shown to transactivate the HIV-1 LTR promoter in a dose-dependent manner when introduced into mammalian cells. In addition, treatment of human endothelial cells with purified Tat proteins resulted in a significant increase in the level of vascular cell adhesion molecule-1 (VCAM-1) expression. These results indicate that the recombinant HIV-1 Tat proteins are active in transactivating viral and cellular promoters. The expression and purification system described in this study will facilitate in characterizing the biological functions of the Tat proteins.

  • PDF

수종 탄닌의 역전사효소와 HIV-1 복제 저해활성 (Inhibitory Activities of Tannins against Reverse Transcriptase and HIV-1 Replication)

  • 김영호;이성우;김항섭;이승호;송만기;성영철;이정준
    • 약학회지
    • /
    • 제39권5호
    • /
    • pp.560-564
    • /
    • 1995
  • Thirty ellagitannins originated from some Euphorbiaceous plants were tested for the inhibitory activities against AMV reverse transcriptase and replication of HIV-1 using syncytia forming assay. Most of ellagitannins showed strong inhibitory activities against AMV reverse transcriptase. Some ellagitannins including geraniin, mallotusinin, elaeocarpusin, euphorscopin and jolkianin, showed significant inhibitory activities of syncytia formation of supT1 cell line.

  • PDF

Inhibitory Constituents against HIV-1 Protease from Agastache rugosa

  • Min, Byung-Sun;Masao-Hattori;Lee, Hyeong-Kyu;Kim, Young-Ho
    • Archives of Pharmacal Research
    • /
    • 제22권1호
    • /
    • pp.75-77
    • /
    • 1999
  • Two diterpenoid compounds, agastanol (1) and agastaquinone (2), were isolated from the roots of Agastache rugosa (Labiatae). Compound 1 and 2 showed significant inhibitory effects against human immunodeficiency virus type 1 (HIV-1) protease activity with $IC_{50}$ values of 360 and $87{\mu}M$, respectively.

  • PDF

Peptide Inhibitor for Human Immunodeficiency Virus Type 1 (HIV-1) Protease from a Thermolysin Hydrolysate of Oyster Proteins

  • Lee, Tae-Gee
    • Fisheries and Aquatic Sciences
    • /
    • 제13권1호
    • /
    • pp.84-87
    • /
    • 2010
  • A peptide that inhibits HIV-1 protease was isolated from a hydrolysate of oyster (Crassostrea gigas) proteins digested with thermolysin. The peptide was using membrane filtration, gel permeation chromatography, ion exchange chromatography, and reverse-phase high performance liquid chromatography. Amino acid sequence of the peptide was determined to be Val-Phe-Glu-Leu. Chemically synthesized Val-Phe-Glu-Leu showed an $IC_{50}$ value of 106 ${\mu}M$.

Effect of Korean Red Ginseng intake on the survival duration of human immunodeficiency virus type 1 patients

  • Cho, Young-Keol;Kim, Jung-Eun
    • Journal of Ginseng Research
    • /
    • 제41권2호
    • /
    • pp.222-226
    • /
    • 2017
  • Background: Long-term ginseng intake can increase longevity in healthy individuals. Here, we examined if long-term treatment with Panax ginseng Meyer (Korean Red Ginseng, KRG) can also enhance survival duration (SD) in patients with human immunodeficiency virus type 1 (HIV-1) infection. Methods: We retrospectively analyzed 252 HIV-1 patients diagnosed from 1986 to 2013 prior to the initiation of antiretroviral therapy. Overall, 162 patients were treated with KRG ($3,947{\pm}4,943g$) for $86{\pm}63$ mo. The effects of KRG on SD were analyzed according to the KRG intake level and the length of the follow-up period. Results: There were significant correlations between the total amount of KRG and SD in the KRG intake group (r = 0.64, p < 0.0001) as well as between total amount of KRG and mean annual decrease in $CD4^+$ T-cell count in all 252 patients (r = -0.17, p < 0.01). The annual decrease in $CD4^+$ T-cell count (change in $cells/{\mu}L$) was significantly slower in KRG-treated patients than in patients receiving no KRG ($48{\pm}40$ vs. $106{\pm}162$; p < 0.001). The SD (in months) was also significantly longer in the KRG group than in the no-KRG group ($101{\pm}64$ vs. $59{\pm}40$, p < 0.01). Conclusion: KRG prolongs survival in HIV-1 patients, possibly by slowing the decrease in $CD4^+$ T-cell count.

Inhibitory Effects of Various Edible Plants and Flavonoids from the Leaves of Cedrela sinensis on Human Immunodeficiency Virus Type 1 Protease

  • Park, Jong-Cheol;Hur, Jong-Moon;Park, Ju-Gwon;Kim, Hyun-Joo;Kang, Kyeong-Hee;Park, Myeong-Rak;Song, Sang-Ho
    • Preventive Nutrition and Food Science
    • /
    • 제5권3호
    • /
    • pp.170-173
    • /
    • 2000
  • The inhibitory effect of extracts from 15 edible plants on the protease of human immunodeficiency virus (HIV) type 1 was investigated. Protease activity was determined by incubating the extracts in a reaction mixture containing protease and substrate His-Lys-Ala-Arg-Val-Leu-(p-NO$_2$-Phe)-Glu-Ala-Nle-Ser-NH$_2$ to inhibit proteolytic cleavage. Of various plants tested, the leaves of Cedrela sinensis inhibited the HIV-1 protease by 42% at a concentration of 100$\mu\textrm{g}$/ml. A major flavonoid isolated from the leaves of C. sinensis, quercetin 3-O-$\alpha$-L-rhamnoside showed inhibitory activity of 19% at a concentration of 100$\mu$M.

  • PDF

Inhibitory Effects of Ninety Nine Korean Plants on Human Immunodeficiency Virus Type 1 Pretense Activity

  • Park, Jong-Cheol;Hur, Jong-Moon;Park, Ju-Gwon;Hyun, Kyu-Hawn;Lee, Kab-Yeon;Hirotsugu Miyashiro;Masao Hattori
    • Preventive Nutrition and Food Science
    • /
    • 제7권2호
    • /
    • pp.123-127
    • /
    • 2002
  • Ninety nine extracts from Korean plants were screened for their inhibitory activities on human immunodeficiency virus (HIV) type 1 pretense by an HPLC method. The pretense inhibitory activities were determined by incubating the extracts in reaction mixtures containing pretense and substrate (His-Lys-Ala-Arg-Val-Leu-(p-NO$_2$- Phe)-Glu-Ala-Nle-Ser-NH$_2$) to perform proteolytic cleavage reactions. Of the extracts tested, the water extracts of Viburnum awabuki (stem and leaves) and Distylium racemosum (leaves) had the highest pretense inhibitory activities at a concentration of 100ug/mL. Activity-guided fractionation, revealed that the n-butanol fraction of the V. awabuki extract and the ethyl acetate fraction from the D. racemosum extract had the greatest inhibitory activity on HIV-1 pretense.

대장균에서 인간면역결핍 바이러스 1형의 gag p24 및 env gp41 단백질의 과발현 및 정제 (Overexpression and Purification of p24 and gp41 Proteins of Human Immunodeficiency Virus Type 1 in E. coli)

  • 김채영;신순천;이성희;김원배;김병문
    • 대한바이러스학회지
    • /
    • 제28권1호
    • /
    • pp.21-30
    • /
    • 1998
  • Synthetic genes encoding the gag p24 and the part of the envelope protein gp41 of the human immunodeficiency virus (HIV-1) were cloned and overexpressed as fusion proteins in Escherichia coli, using an expression vector carrying T7 promoter and the poly-histidine leader, sequence. The overexpressed p24 fusion protein was purified by centrifugation, Ni-affinity chromatography and CM-sepharose chromatography. The overexpressed gp41 fusion protein was purified by centrifugation, $C_4$ chromatography and DEAE-sepharose chromatography. The purified fusion proteins showed a high level of purity and immunoreactivity in SDS-polyacrylamide gel electrophoresis and western blot analysis. These results suggest that this prokaryotic - expression purification method is suitable for obtaining a large amount of the viral antigen which may be useful for screening of antibodies to HIV-1 in human blood samples.

  • PDF

이식을 위한 사람 양막의 소독 및 멸균공정에 의한 감염성 위해인자 불활화 효과 (Inactivation of Infectious Microorganisms by Disinfection and Sterilization Processes for Human Amniotic Membrane Grafts)

  • 배정은;김찬경;김인섭
    • 미생물학회지
    • /
    • 제45권4호
    • /
    • pp.346-353
    • /
    • 2009
  • 이식을 위해 사용하는 사람 양막은 기증자로부터 수혜자에게 바이러스, 세균, 진균과 같은 감염성 위해인자를 전파할 위험이 있다. 따라서 적절한 소독 및 멸균 공정을 통해 이식용 양막 내재 또는 혼입 가능한 감염성 위해인자를 완벽하게 불활화하여야 한다. 본 연구에서는 인체조직은행에서 사용하고 있는 소독 공정과 멸균 공정의 바이러스 및 세균, 진균 불활화 효과를 검증하기 위해 국제적 가이드에 따라 5종의 바이러스[human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), porcine parvovirus (PPV)]와 2종의 세균(Escherichia coli, Bacillus subtilis), 1종의 진균(Candida albicans)을 생물학적 지표로 사용하였다. 양막에 각 생물학적 지표를 첨가한 후 70% 에탄올 소독 공정, 감마선 조사 공정, 산화에틸렌 가스 멸균 공정을 실시한 다음 각 바이러스, 세균, 진균을 회수하여 정량한 후 불활화 정도를 비교하였다. 70% 에탄올 처리 공정에서 HIV-1, BHV, BVDV 같은 외피 바이러스는 처리 시간 2.5분 안에 불활화되었지만, HAV와 PPV 같은 비-외피 바이러스는 에탄올에 매우 큰 저항성을 나타내었다. 감마선 2.5 kGy 조사에 의해 HIV-1, BHV, BVDV는 검출한계 이하로 완벽하게 불활화되었다. HAV와 PPV는 각각 5 kGy와 25 kGy 조사에 의해 검출한계 이하로 불활화되었다. 산화에틸렌 가스 처리에 의해 본 연구에 사용한 모든 바이러스가 검출한계 이하로 불활화되었다. 70% 에탄올 처리 공정에서 E. coli와 C. albicans는 모두 5분 안에 완벽하게 사멸하였다. 하지만 B. subtilis는 큰 저항성을 나타내었다. 감마선 조사 공정과 산화에틸렌 가스 멸균 공정에서 E. coli, B. subtilis, C. albicans 모두 완벽하게 불활화되었다.