• Title/Summary/Keyword: HIV-1 system

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Development and Utilization of an Human Immunodeficiency Virus Type 1 (HIV-1)-Based Retroviral Vector System to Express HIV-1 Envelope Glycoprotein in $CD4^+$ T Cells (HIV-1 Retroviral Vector System의 개발 및 $CD4^+$ T 세포에서 HIV-1 Envelope 당단백질의 발현)

  • Park, Jin-Seu
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.141-150
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    • 1996
  • 본 연구에서는 $CD4^+$ T 세포를 일회성으로 감염시킬 수 있고 $CD4^+$ T 세포에 HIV-1 envelope 유전자를 전달할 수 있는 바이러스 입자를 생산하는 HIV-1 complementation system을 개발하였고 이 system을 이용하여 $CD4^+$ T 세포에 선택적으로 HIV-1 envelope 당단백질을 발현시켰다. 이 system은 Gag/Gag-Pol expressor와 Env expressor로 구성되어있다. Gag/Gag-Pol expressor는 바이러스 입자 생산에 필요한 구조단백질과 기능단백질을 발현시키지만 packaging signal이 결핍되어 바이러스 입자로 유전자가 들어가지 못하도록 제조되었다. Env expressor는 Tat, Rev와 envelope 당단백질을 발현시키고 packaging signal을 갖고 있어 바이러스 입자로 envelope 유전자가 들어가도록 제조되었다. Gag/Gag-Pol expressor로부터 Gag와 Gag-Pol의 발현은 Rev 단백질을 요구하였고 Env expressor로부터 Rev 단백질 이 제공될 때 Gag와 Gag-Pol 단백질은 효율적으로 발현되었다. Gag/Gag-Pol과 Env expressor로 cotransfection된 COS-1 세포에서 $CD4^+$ T 세포를 일회성으로 감염시킬 수 있는 바이러스 입자가 생산되었다. 생산된 바이러스 입자는 $CD4^+$ T 세포에 HIV-1 envelope 유전자를 전달하여 envelope 당단백질을 발현시켰고 복제 가능한 자손 바이러스의 생성을 유도하지 못하였다. 본 연구에서 개발된 방법은 $CD4^+$ T 세포에서 envelope 당단백질의 기능을 분석하고 관심 있는 유전자를 $CD4^+$ T 세포에 전달하는 바이러스 입자의 생산에 이용할 수 있다.

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Application of HIV-1 Complementation System to Screen the Anti-AIDS Agents That Targets the Late Stage of HIV-1 Replication Cycle (바이러스 생활환의 후기 단계에 작용하는 항AIDS제의 탐색을 위한 HIV-1 Complementation System의 응용)

  • Ryu, Ji-Yoon;Choi, Soo-Young;Kim, Yung-Hi;Park, Jin-Seu
    • The Journal of Korean Society of Virology
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    • v.30 no.3
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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.

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Isolation of the Gene for HIV-1 gp41 Interacting Protein (HIV gp41의 세포내 부분과 상호작용하는 단백질 유전자의 분리)

  • Kim, Eun-Mi;Kim, Jung-Woo
    • The Journal of Natural Sciences
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    • v.10 no.1
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    • pp.27-32
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    • 1998
  • To find the interacting protein with the cytoplasmic domain of HIV-1 gp41, the yeast two hybrid system was used for the expression cloning. Among the $1.4 \times 10^6 colonies, 20 colonies were selected as the final candidate for the interacting protein gene. The nucleotide sequencing revealed three kinds of protein, acidic ribosomal protein P0, beta tubulin, alpha catenin. These proteins interacted with the gp41 specifically in yeast system.

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Mature HIV-like Particles Produced from Single Semliki Forest Virus-Derived Expression Vector

  • KIM EUN;POO HAR-YOUNG;SUNG MOON-HEE;KIM CHUL-JOONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1229-1239
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    • 2005
  • Human immunodeficiency virus-like particles (HIVVLPs) with native conformations similar to that of the wild-type virion could be valid candidates for vaccine development. To this end, we used a Semliki Forest Virus (SFV) expression system to produce HIV- VLPs containing high quantities of native envelope proteins. Here, we described a single SFV replicon containing the HIV gagpol and env genes under the control of separate subgenomic promoters. Mature VLPs incorporating the Gag and Env proteins were detected in the supernatant of replicon-expressing cells by Western blot analysis. The HIV-VLPs showed the expected molecular density (1.14-1.18 g/ml) on a $20-60\%$ sucrose gradient; the particles were 100-120 nm in diameter and Env proteins were observed on their surfaces by immunogold electron microscopy. RT-PCR analysis of VLP-associated RNAs in mature HIV-VLPs revealed two SF V-derived RNA species (full-length and subgenomic). Immunization studies in Balb/c mice showed that these HIV-VLPs were capable of inducing both HIV-specific antibodies and cell-mediated immune responses. Taken together, our results indicate that the SFV replicon system is useful for the production of HIV-VLPs, which may be valuable candidates for an HIV vaccine.

Anti-HIV-1 Activity of Gelatin Hydrolysate Derived from Alaska Pollack Theragra chalcogramma Skin (명태(Theragra chalcogramma) 껍질 유래 젤라틴 가수분해물의 항 HIV-1 효능)

  • Park, Sun-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.594-599
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    • 2016
  • Infection with HIV (Human immunodeficiency virus), over time, develops into acquired immunodeficiency syndrome (AIDS). The development of non-toxic and effective anti-HIV drugs is one of the most promising strategies for the treatment of AIDS. In this study, we investigated the anti-HIV-1 activity of gelatin hydrolysates from Alaska pollack skin. Gelatin hydrolysates were prepared using four enzymes (alcalase, flavourzyme, neutrase, and pronase E). Among these, the pronase E gelatin hydrolysate was found to inhibit HIV-1 infection in the human T cell-line MT4. It exhibited inhibitory activity on HIV-1IIIB-induced cell lysis, reverse transcriptase activity, and viral p24 production at noncytotoxic concentrations. Moreover, it decreased the activation of matrix metalloproteinase-2 (MMP-2) in vitro. Because HIV infection-induced activation of MMP-2 can accelerate collagen resolution and collapse of the immune system, pronase E gelatin hydrolysate might prevent the activation of MMP-2 in cells, resulting in collagen stabilization and immune cell homeostasis consistent with anti-HIV activation. These results suggest that pronase E gelatin hydrolysate could potentially be incorporated into a novel therapeutic agent for HIV/AIDS patients.

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
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    • v.33 no.4
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    • pp.337-343
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    • 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.

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ESTIMATING THE DOMAIN OF ATTRACTION OF HIV-1 SYSTEM BASED ON MOMENT METHOD

  • LI, CHUNJI;YAO, HAN
    • Journal of applied mathematics & informatics
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    • v.36 no.5_6
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    • pp.349-358
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    • 2018
  • In this article, we discuss the domain of attraction of HIV-1 system by using the moment theory. First, the asymptotic stabilities of the equilibrium point of the system are given, and then we introduce how to use the moment method to estimate domain of attraction. Finally, one simulation shows the effectiveness of moment method.

STABILIZATION OF HIV / AIDS MODEL BY RECEDING HORIZON CONTROL

  • ELAIW A. M.;KISS K.;L CAETANO M. A.
    • Journal of applied mathematics & informatics
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    • v.18 no.1_2
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    • pp.95-112
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    • 2005
  • This work concerns the stabilization of uninfected steady state of an ordinary differential equation system modeling the interaction of the HIV virus and the immune system of the human body. The control variable is the drug dose, which, in turn, affects the rate of infection of $CD4^{+}$ T cells by HIV virus. The feedback controller is constructed by a variant of the receding horizon control (RHC) method. Simulation results are discussed.

Prevalence of Peripheral Vestibular Impairment in Adults with Human Immunodeficiency Virus

  • Millar, Alison;Joubert, Karin;Naude, Alida
    • Korean Journal of Audiology
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    • v.25 no.1
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    • pp.36-42
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    • 2021
  • Background and Objectives: Globally, the human immunodeficiency virus (HIV) is responsible for one of the most serious pandemics to date. The vulnerability of the vestibular system in individuals with HIV has been confirmed, and central vestibular impairments have been frequently reported. However, there are disagreements on the impact of HIV on peripheral vestibular function. Thus, the current study aimed to determine the prevalence of peripheral vestibular impairment, specifically related to the semi-circular canals (SCCs), in HIV-positive individuals receiving antiretroviral (ARV) treatment. Subjects and Methods: A total of 92 adults between the ages of 18 and 50 years (divided into two groups) participated in the study. The first group comprised HIV-positive individuals receiving ARV treatment (n1=60), and the second group comprised HIV-negative participants (n2=32). The video head impulse test was used to conduct the head impulse paradigm (HIMP). Results: Bilateral normal HIMP results were obtained in 95% of the HIV-positive participants and all HIV-negative participants. The gain of the left posterior SCCs was significantly lower in the HIV-positive group, while the gains of all other canals between the two groups were comparable. Conclusions: The prevalence of peripheral vestibular impairment in the HIV-positive group was not significantly different from that of the HIV-negative group. The reduced prevalence in the current study may be attributed to participant characteristics, the test battery employed, and the central compensation of the vestibular dysfunctions at the later stages of infection.

Prevalence of Peripheral Vestibular Impairment in Adults with Human Immunodeficiency Virus

  • Millar, Alison;Joubert, Karin;Naude, Alida
    • Journal of Audiology & Otology
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    • v.25 no.1
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    • pp.36-42
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    • 2021
  • Background and Objectives: Globally, the human immunodeficiency virus (HIV) is responsible for one of the most serious pandemics to date. The vulnerability of the vestibular system in individuals with HIV has been confirmed, and central vestibular impairments have been frequently reported. However, there are disagreements on the impact of HIV on peripheral vestibular function. Thus, the current study aimed to determine the prevalence of peripheral vestibular impairment, specifically related to the semi-circular canals (SCCs), in HIV-positive individuals receiving antiretroviral (ARV) treatment. Subjects and Methods: A total of 92 adults between the ages of 18 and 50 years (divided into two groups) participated in the study. The first group comprised HIV-positive individuals receiving ARV treatment (n1=60), and the second group comprised HIV-negative participants (n2=32). The video head impulse test was used to conduct the head impulse paradigm (HIMP). Results: Bilateral normal HIMP results were obtained in 95% of the HIV-positive participants and all HIV-negative participants. The gain of the left posterior SCCs was significantly lower in the HIV-positive group, while the gains of all other canals between the two groups were comparable. Conclusions: The prevalence of peripheral vestibular impairment in the HIV-positive group was not significantly different from that of the HIV-negative group. The reduced prevalence in the current study may be attributed to participant characteristics, the test battery employed, and the central compensation of the vestibular dysfunctions at the later stages of infection.