3.HIV-1 infection up-regulating expression of interferon-stimulated gene 15 in cell lines.
Huan-mei WU ; Jun SUN ; Zhe-feng MENG ; Xiao-yan ZHANG ; Jian-qing XU
Chinese Journal of Virology 2013;29(5):480-487
To investigate whether HIV-1 infection affects expression of interferon-stimulated gene 15 (ISG15) and determine the antiviral effect of ISG15 in vitro, ISG15 expression at transcription and protein level and supernatant p24 of HIV-1 was detected in various HIV-1 infected or transfected cell lines, respec tively. HIV-1 molecular clone pNL4-3 was used to transfect 293T, TZM-bl and HeLa cells while HIV-1 pseudo-typed virus was used to infect Jurkat, MT-1 and THP-1 cells. After twenty-four hours post infection or transfection, cells were harvested for extraction of total RNAs and subsequently used in real time PCR for quantification of ISG15 transcriptional expression. After forty-eight hours post infection or transfection, cells were harvested for extraction of total proteins to detect ISG15 protein expression. A significant up-regulation of ISG15 at transcription level was observed in HIV-1 infected or transfected cell lines, particulaly in THP-1 and TZM-bl cells. Up-regulation of ISG15 protein was observed only in TZM-bl cell. Cotransfection of ISG15 and HIV-1 indicated that ISG15 inhibited production of HIV-1 progeny virus in a dose and time depend manner in 293T cell but not TZM-bl cell. These results revealed upregulating ISG15 expression in transcriptional level and potential antagonistic mechanism against ISG15 by HIV-1 infection, simultanelusly.
Base Sequence
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Cell Line
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Cytokines
;
genetics
;
metabolism
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HIV Infections
;
genetics
;
metabolism
;
virology
;
HIV-1
;
physiology
;
Humans
;
Interferons
;
metabolism
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Molecular Sequence Data
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Ubiquitins
;
genetics
;
metabolism
;
Up-Regulation
4.An atlas of immune cell transcriptomes in human immunodeficiency virus-infected immunological non-responders identified marker genes that control viral replication.
Yahong CHEN ; Xin LI ; Shuran LIU ; Wen AO ; Jing LIN ; Zhenting LI ; Shouli WU ; Hanhui YE ; Xiao HAN ; Dongliang LI
Chinese Medical Journal 2023;136(22):2694-2705
BACKGROUND:
Previous studies have examined the bulk transcriptome of peripheral blood immune cells in acquired immunodeficiency syndrome patients experiencing immunological non-responsiveness. This study aimed to investigate the characteristics of specific immune cell subtypes in acquired immunodeficiency syndrome patients who exhibit immunological non-responsiveness.
METHODS:
A single-cell transcriptome sequencing of peripheral blood mononuclear cells obtained from both immunological responders (IRs) (CD4 + T-cell count >500) and immunological non-responders (INRs) (CD4 + T-cell count <300) was conducted. The transcriptomic profiles were used to identify distinct cell subpopulations, marker genes, and differentially expressed genes aiming to uncover potential genetic factors associated with immunological non-responsiveness.
RESULTS:
Among the cellular subpopulations analyzed, the ratios of monocytes, CD16 + monocytes, and exhausted B cells demonstrated the most substantial differences between INRs and IRs, with fold changes of 39.79, 11.08, and 2.71, respectively. In contrast, the CD4 + T cell ratio was significantly decreased (0.39-fold change) in INRs compared with that in IRs. Similarly, the ratios of natural killer cells and terminal effector CD8 + T cells were also lower (0.37-fold and 0.27-fold, respectively) in the INRs group. In addition to several well-characterized immune cell-specific markers, we identified a set of 181 marker genes that were enriched in biological pathways associated with human immunodeficiency virus (HIV) replication. Notably, ISG15 , IFITM3 , PLSCR1 , HLA-DQB1 , CCL3L1 , and DDX5 , which have been demonstrated to influence HIV replication through their interaction with viral proteins, emerged as significant monocyte marker genes. Furthermore, the differentially expressed genes in natural killer cells were also enriched in biological pathways associated with HIV replication.
CONCLUSIONS
We generated an atlas of immune cell transcriptomes in HIV-infected IRs and INRs. Host genes associated with HIV replication were identified as markers of, and were found to be differentially expressed in, different types of immune cells.
Humans
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Acquired Immunodeficiency Syndrome
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Transcriptome/genetics*
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HIV
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HIV Infections/genetics*
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Leukocytes, Mononuclear/metabolism*
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CD4-Positive T-Lymphocytes/metabolism*
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Virus Replication
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Membrane Proteins/metabolism*
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RNA-Binding Proteins/metabolism*
5.CCR5, a new target of anti-HIV drugs.
Yan-xing HAN ; Jian-dong JIANG
Acta Academiae Medicinae Sinicae 2003;25(5):635-639
CCR5, a membrane protein on cell surface, is a member of the G protein-coupled receptor superfamily and one of the major co-receptors for HIV-1 infection. The roles of CCR5 in HIV-1 infection have been elucidated since 1996. Because of the biological nature of CCR5, it has became a molecular target for the novel drugs against HIV-1. Antagonists for CCR5 could be grouped as following, chemokine derivatives, small molecule non-peptide compounds, monoclonal antibodies and peptides. The latest progress in this field is reviewed in this article.
Anti-HIV Agents
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pharmacology
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Antibodies, Monoclonal
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CCR5 Receptor Antagonists
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Drug Design
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HIV Infections
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metabolism
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HIV-1
;
drug effects
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Receptors, CCR5
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drug effects
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Receptors, Chemokine
;
drug effects
6.A transcriptional study of SDF-1alpha expression in PBL and the correlation between SDF-1alpha transcription and HIV-1 infection.
Xiu-Ying ZHAO ; Rui-Shan LI ; Jia-Qing HUANG ; Zhi-Wei CHEN ; Bo-Jian ZHENG
Chinese Journal of Experimental and Clinical Virology 2009;23(3):204-207
OBJECTIVETo determine the transcription of SDF-1alpha in peripheral blood lymphocytes (PBL) and analysis the correlation between SDF-1alpha transcription and HIV infection.
METHODSThree groups of study subjects were recruited: (1) 97 HIV negative healthy donors, (2) 92 HIV patients of A1 to A3 stages and (3) 146 HIV patients of B1 to C3 stages. Total RNA was extracted from PBL. Reverse transcription (RT)-PCR and quantification PCR were developed for the SDF-1alpha transcriptional study. R1 value was calculated based on the ratio of SDF-1alpha copies to beta-globin copies.
RESULTSSDF-1alpha transcription is heterogeneous among the three study groups. The SDF-1alpha transcription was significantly up-regulated during late stage of HIV infection than the healthy donors. Correlation analysis indicated that R1 value was negatively correlated to CD4+ T cells counts (P = 0.002); and positively correlated to virus load (P = 0.001). The result demonstrated an association between SDF-1alpha transcription and disease progression.
CONCLUSIONSDF-1alpha transcription was significantly up-regulated during late stage of HIV infection. It would be worthwhile to determine the mechnism of HIV affecting on SDF-1alpha genes transcription and the up-regulated SDF-1alpha expression on the disease progression.
Case-Control Studies ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; metabolism ; HIV Infections ; genetics ; metabolism ; virology ; HIV-1 ; genetics ; physiology ; Humans ; Lymphocytes ; metabolism ; virology ; Transcription, Genetic ; Up-Regulation
7.Characterization of Gp41 Polymorphisms in the Fusion Peptide Domain and T-20 (Enfuvirtide) Resistance-Associated Regions in Korean HIV-1 Isolates.
Dai Ho JANG ; Cheol Hee YOON ; Byeong Sun CHOI ; Yoon Seok CHUNG ; Hye Young KIM ; Sung Gil CHI ; Sung Soon KIM
Journal of Korean Medical Science 2014;29(3):456-459
HIV-1 gp41 is an envelope protein that plays an essential role in virus entry. The mutation of gp41 affects HIV-1 entry and susceptibility to the fusion inhibitor T-20. Therefore, we analyzed the natural polymorphism of gp41 of 163 HIV-1 isolates from T-20-naive Koreans infected with HIV-1. This study of gp41 polymorphisms showed that insertions in the fourth threonine (74.8%) and L7M substitutions (85.3%) were more frequent in the fusion peptide motif in Korean HIV-1 isolates compared with those from other countries. Minor T-20 resistance mutations such as L45M (1.2%), N126K (1.2%), and E137K (6.7%) were detected, but the critical T-20 resistance mutations were not detected in the gp41 HR1 and HR2 regions. In addition, the N42S mutation (12.9%) associated with T-20 hypersusceptibility was detected at a high frequency. These results may serve as useful data for studies considering T-20 for use in the development of a more effective anti-retroviral treatment in Korea.
Anti-HIV Agents/pharmacology
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Drug Resistance, Viral/*genetics
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HIV Envelope Protein gp41/*genetics/metabolism/pharmacology
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HIV Infections/virology
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HIV-1/*genetics/isolation & purification/*metabolism
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Humans
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Peptide Fragments/pharmacology
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*Polymorphism, Genetic
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Protein Structure, Tertiary/genetics
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Republic of Korea
;
Virus Internalization
8.Establishment of stable cell line and expression and purification of HIV gp120 in Drosophila S2 cells.
Pei-Long SUN ; Jian-Hua ZHOU ; Shu-Xia LV ; Xin-Qi LIU
Chinese Journal of Virology 2010;26(6):460-464
Constructing eukaryotic expressing vector of pMT/Bip/V5-His A-HIV gp120 and transfecting S2 cells to establish stable gp120-expressing S2 cell line. The gp120 gene of HIV CRF07-BC epidemic in China was amplified by polymerase chain reaction from the recombinant vector PRSV-gp120 and confirmed by DNA sequence analysis. The DNA fragment of HIV gp120 was digested with the restriction endonucleases NcoI and XhoI, then inserted into eukaryotic expressing vector pMT/BiP/V5-His A. A selection vector containing the streptomyces griseochromogenes bsd gene conferring blasticidin resistance was cotransfected into drosophila S2 cells by Cellfectin II reagent. The stable cell line was established following repeated screening by blasticidin. HIV gp120 was purified by a Ni-NTA affinity column followed by molecular sieve chromatography. Recombinant protein was characterized by SDS-PAGE, Western blot and ELISA. The eukaryotic expressing vector pMT/BiP/V5-His A-HIV gp120 was constructed, stable expressing cell line was established, and protein was expressed and purified successfully. This result will contribute to functional study of gp120 and vaccine design against AIDS.
Animals
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Cell Line
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Drosophila
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genetics
;
metabolism
;
virology
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Gene Expression
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HIV
;
genetics
;
metabolism
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HIV Envelope Protein gp120
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genetics
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isolation & purification
;
metabolism
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HIV Infections
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virology
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Humans
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Recombinant Proteins
;
genetics
;
isolation & purification
;
metabolism
9.Advances in the study of molecular mechanism of APOBEC3G anti-HIV-1.
Bo FAN ; Shan CEN ; Jian-dong JIANG
Acta Pharmaceutica Sinica 2008;43(7):678-682
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 protein G (APOBEC3G) is part of the innate immune system of host cells and has cytidine deaminase activity. It specifically incorporates into the virion during HIV-1 replication. The incorporation of APOBEC3G needs its interaction with HIV-1 Gag. In the HIV-1 reverse transcription process, APOBEC3G deaminates dC to dU in the first minus strand cDNA, and then induces extensive hypermutation in the viral genome. Besides deamination, APOBEC3G also inhibits HIV-1 by some kinds of non-deamination mechanisms which need to be further elucidated. HIV-1 Vif counteracts the activity of APOBEC3G by an ubiquitin-proteasome-mediated degradation of APOBEC3G. As a broad spectrum inhibitor of viruses, APOBEC3G also inhibits various retroviruses, retrotransposons and other viruses like HBV. Upregulating the expression of APOBEC3G or blocking the Vif-mediated degradation of APOBEC3G might be novel strategies to treat HIV-1 infection in the future.
APOBEC-3G Deaminase
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Amino Acid Substitution
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Anti-HIV Agents
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metabolism
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Cytidine Deaminase
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genetics
;
metabolism
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Gene Expression
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HIV Infections
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metabolism
;
HIV-1
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genetics
;
physiology
;
Humans
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Virus Replication
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vif Gene Products, Human Immunodeficiency Virus
;
genetics
;
metabolism
10.Effect of HIV Tat protein on CCR5 expression in monocytes and infection with monocyte-tropic HIV strains.
Yi-da YANG ; Lin ZHENG ; Guo-cai LU ; Ya-gang CHEN ; Maria SALVATO
Journal of Zhejiang University. Medical sciences 2004;33(6):532-545
OBJECTIVETo study the effects of HIV Tat protein on CCR5 expression of monocytes and HIV infection in monocytes.
METHODSMembrane expression of CCR5 on monocytes was analyzed by flow cytometry. Stimulated with HIV Tat protein, monocytes were infected with monocyte-tropic HIV(Ba-L) and HIV gag p24 level in the supernatant was measured by ELISA methods.
RESULTSHIV Tat protein increased CCR5 expression in human monocytes,which was inhibited by rabbit anti-Tat polyclonal antibody. Tat protein also increased p24 level after monocyte-tropic HIV-1(Ba-L) infected monocytes.
CONCLUSIONTat increases CCR5 expression and HIV-1 infection in monocytes, which indicates that HIV Tat might be a key protein in HIV-1 infection.
Flow Cytometry ; Gene Products, tat ; pharmacology ; HIV ; HIV Infections ; metabolism ; Humans ; Monocytes ; metabolism ; Receptors, CCR5 ; biosynthesis ; genetics ; tat Gene Products, Human Immunodeficiency Virus