Cytokine Signals Are Sufficient for HIV-1 Infection of Resting Human T Lymphocytes

Lentiviral vectors have been advocated to be effective vehicles for the delivery and stable expression of genes in nondividing primary cells. However, certain cell types, such as resting T lymphocytes, are resistant to infection with HIV-1. Establishing parameters for stable gene delivery into primary human lymphocytes and approaches to overcome the resistance of resting T cells to HIV infection may permit potential gene therapy applications, genetic studies of primary cells in vitro, and a better understanding of the stages of the lentiviral life cycle. Here we demonstrate that an HIV-1–derived vector can be used for stable delivery of genes into activated human T cells as well as natural killer and dendritic cells. Remarkably, a sizeable fraction of resting T cells was stably transduced with the HIV-1 vector when cultured with the cytokine interleukin (IL)-2, IL-4, IL-7, or IL-15, or, at a lower level, with IL-6, in the absence of any other stimuli. Resting T cells stimulated with these cytokines could also be infected with replication-competent HIV-1. To test the utility of this system for performing structure–function analysis in primary T cells, we introduced wild-type as well as a mutant form of murine CD28 into human T cells and showed a requirement for the CD28 cytoplasmic domain in costimulatory signaling. The ability to stably express genes of interest in primary T cells will be a valuable tool for genetic and structure–function studies that previously have been limited to transformed cell lines. In addition, the finding that cytokine signals are sufficient to permit transduction of resting T cells with HIV may be relevant for understanding mechanism of HIV-1 transmission and pathogenesis.

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Lipocortin 1 binding sites on human T-cells: The population of cells with the binding sites is larger in CD8 T-lymphocytes than in CD4 T-lymphocytes

IUBMB Life ◽

10.1080/15216549600201803 ◽

1996 ◽

Vol 40 (6) ◽

pp. 1167-1173

Author(s):

Ha Won Kim ◽

Euna Choi ◽

Bin Yoo ◽

Jung Ryul Choi ◽

Young Min Park ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Binding Sites ◽

Cd8 T Lymphocytes ◽

Human T Cells

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Disruption of HIV-1 Co-Receptors CCR5 and CXCR4 in Primary Human T Cells and Hematopoietic Stem and Progenitor Cells Using Base Editing

Molecular Therapy ◽

10.1016/j.ymthe.2021.10.026 ◽

2021 ◽

Author(s):

Friederike Knipping ◽

Gregory A. Newby ◽

Cindy R. Eide ◽

Amber N. McElroy ◽

Sarah C. Nielsen ◽

...

Keyword(s):

T Cells ◽

Progenitor Cells ◽

Hematopoietic Stem ◽

Base Editing ◽

Human T Cells ◽

Stem And Progenitor Cells ◽

Hiv 1

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Generation of primary antigen-specific human cytotoxic T lymphocytes in human/mouse radiation chimera

Blood ◽

10.1182/blood.v88.2.721.bloodjournal882721 ◽

1996 ◽

Vol 88 (2) ◽

pp. 721-730 ◽

Cited By ~ 30

Author(s):

H Segall ◽

I Lubin ◽

H Marcus ◽

A Canaan ◽

Y Reisner

Keyword(s):

T Cells ◽

T Cell ◽

Human Lymphocytes ◽

Scid Mice ◽

Human Antibody ◽

Adoptive Therapy ◽

Activation Markers ◽

Human T Cell ◽

Human T Cells

Severe combined immunodeficient (SCID) mice are increasingly used as hosts for the adoptive transfer of human lymphocytes. Human antibody responses can be obtained in these xenogeneic chimeras, but information about the functionality of the human T cells in SCID mice is limited and controversial. Studies using human peripheral blood lymphocytes (PBL) injected intraperitoneally (IP) into SCID mice (hu-PBL-SCID mice) have shown that human T cells from these chimeras are anergic and have a defective signaling via the T-cell receptor. In addition, their antigenic repertoire is limited to xenoreactive clones. In the present study, we tested the functionality of human T cell in a recently described chimeric model. In this system, BALB/c mice are conditioned by irradiation and then transplanted with SCID bone marrow, followed by IP injection of human PBL. Our experiments demonstrated that human T cells, recovered from these hu-PBL-BALB mice within 1 month posttransplant, proliferated and expressed activation markers upon stimulation with anti-CD3 monoclonal antibody. A vigorous antiallogeneic human cytotoxic T-lymphocyte (CTL) response could be generated in these mice by immunizing them with irradiated allogeneic cells. Moreover, anti-human immunodeficiency virus type 1 (HIV-1) Net- specific human CTLs could be generated in vivo from naive lymphocytes by immunization of mouse-human chimeras with a recombinant vaccinia-nef virus. This model may be used to evaluate potential immunomodulatory drugs or cytokines, and could provide a relevant model for testing HIV vaccines, for production of antiviral T-cell clones for adoptive therapy, and for studying human T-cell responses in vivo.

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Potent inhibition of HIV-1 gene expression and TAT-mediated apoptosis in human T cells by novel mono- and multitarget anti-TAT/Rev/Env ribozymes and a general purpose RNA-cleaving DNA-enzyme

Antiviral Research ◽

10.1016/j.antiviral.2006.05.009 ◽

2006 ◽

Vol 72 (2) ◽

pp. 134-144 ◽

Cited By ~ 12

Author(s):

Hoshang Unwalla ◽

Samitabh Chakraborti ◽

Vikas Sood ◽

Nidhi Gupta ◽

Akhil C. Banerjea

Keyword(s):

Gene Expression ◽

T Cells ◽

General Purpose ◽

Human T Cells ◽

Potent Inhibition ◽

Hiv 1

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Synthesis, Characterization and anti-HIV and Antitumor Activities of New Coumarin Derivatives

Zeitschrift für Naturforschung B ◽

10.1515/znb-2008-0112 ◽

2008 ◽

Vol 63 (1) ◽

pp. 83-89 ◽

Cited By ~ 28

Author(s):

Yaseen A. Al-Soud ◽

Haitham H. Al-Sa’doni ◽

Houssain A. S. Amajaour ◽

Kifah S. M. Salih ◽

Mohammad S. Mubarakb ◽

...

Keyword(s):

T Cells ◽

In Vitro Cytotoxicity ◽

Tumor Cell Lines ◽

Coumarin Derivatives ◽

Antitumor Activities ◽

Reverse Transcriptase Inhibitors ◽

Human T Cells ◽

Anti Hiv ◽

Hiv 1

A new series of coumarin and benzofuran derivatives were synthesized as potential non-nucleoside reverse transcriptase inhibitors (NNRTIs) by reacting, separately, 4-bromomethylcoumarins, their sulphonyl chlorides, and ethyl 3-(bromomethyl)-6-methoxy-1-benzofuran-2-carboxylate with different imidazoles and their benzo analogs. The antiviral (HIV-1, HIV-2) properties of the newly synthesized compounds were investigated in vitro and all compounds were found to be inactive, except 10 which showed inhibition of HIV-2 with EC50 > 0.51 μgmL−1. The in vitro cytotoxicity of 17 and 19 was assayed against a panel of tumor cell lines consisting of CD4 human T-cells.

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Rev and the fate of pre-mRNA in the nucleus: implications for the regulation of RNA processing in eukaryotes

Molecular and Cellular Biology ◽

10.1128/mcb.13.10.6180-6189.1993 ◽

1993 ◽

Vol 13 (10) ◽

pp. 6180-6189 ◽

Cited By ~ 6

Author(s):

M H Malim ◽

B R Cullen

Keyword(s):

T Cells ◽

Regulation Of Gene Expression ◽

Nucleocytoplasmic Transport ◽

Viral Mrnas ◽

Nuclear Retention ◽

Human T Cells ◽

Cellular Factors ◽

Immunodeficiency Virus ◽

Nuclear Degradation ◽

Hiv 1

Although a great deal is known about the regulation of gene expression in terms of transcription, relatively little is known about the modulation of pre-mRNA processing. In this study, we exploited a genetically regulated system, human immunodeficiency virus type 1 (HIV-1) and its trans-activator Rev, to examine events that occur between the synthesis of pre-mRNA in the nucleus and the translation of mRNA in the cytoplasm. Unlike the majority of eukaryotic pre-mRNAs whose introns are efficiently recognized and spliced prior to nucleocytoplasmic transport, HIV-1 mRNAs containing functional introns must be exported to the cytoplasm for the expression of many viral proteins. Using human T cells containing stably integrated proviruses, we demonstrate that such incompletely spliced viral mRNAs are exported to the cytoplasm only in the presence of the Rev trans-activator. In the absence of Rev, these intron-containing RNAs are sequestered in the T-cell nucleus and either spliced or, more commonly, degraded. Because Rev does not inhibit the expression of fully spliced viral mRNA species in T cells, we propose that Rev, rather than inhibiting viral pre-mRNA splicing, is acting here both to prevent the nuclear degradation of HIV-1 pre-mRNAs and to induce their translocation to the cytoplasm. Taken together, these findings indicate that the cellular factors responsible for the nuclear retention of unspliced pre-mRNAs, although most probably splicing factors, do not invariably commit these RNAs to productive splicing and can, instead, program such transcripts for degradation.

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Interferon- Activates Multiple STAT Proteins and Upregulates Proliferation-Associated IL-2R, c-myc, and pim-1 Genes in Human T Cells

Blood ◽

10.1182/blood.v93.6.1980.406k20_1980_1991 ◽

1999 ◽

Vol 93 (6) ◽

pp. 1980-1991 ◽

Cited By ~ 186

Author(s):

Sampsa Matikainen ◽

Timo Sareneva ◽

Tapani Ronni ◽

Anne Lehtonen ◽

Päivi J. Koskinen ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Lymphocytes ◽

Cell Biology ◽

Interleukin 2 ◽

T Cell Proliferation ◽

Stat Proteins ◽

Human T Cells ◽

T Cell Biology ◽

Cytokine Stimulation

Interferon- (IFN-) is a pleiotropic cytokine that has antiviral, antiproliferative, and immunoregulatory functions. There is increasing evidence that IFN- has an important role in T-cell biology. We have analyzed the expression ofIL-2R, c-myc, and pim-1 genes in anti-CD3–activated human T lymphocytes. The induction of these genes is associated with interleukin-2 (IL-2)–induced T-cell proliferation. Treatment of T lymphocytes with IFN-, IL-2, IL-12, and IL-15 upregulated IL-2R, c-myc, andpim-1 gene expression. IFN- also sensitized T cells to IL-2–induced proliferation, further suggesting that IFN- may be involved in the regulation of T-cell mitogenesis. When we analyzed the nature of STAT proteins capable of binding to IL-2R,pim-1, and IRF-1 GAS elements after cytokine stimulation, we observed IFN-–induced binding of STAT1, STAT3, and STAT4, but not STAT5 to all of these elements. Yet, IFN- was able to activate binding of STAT5 to the high-affinity IFP53 GAS site. IFN- enhanced tyrosine phosphorylation of STAT1, STAT3, STAT4, STAT5a, and STAT5b. IL-12 induced STAT4 and IL-2 and IL-15 induced STAT5 binding to the GAS elements. Taken together, our results suggest that IFN-, IL-2, IL-12, and IL-15 have overlapping activities on human T cells. These findings thus emphasize the importance of IFN- as a T-cell regulatory cytokine.

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Telomerase levels control the lifespan of human T lymphocytes

Blood ◽

10.1182/blood-2002-07-2015 ◽

2003 ◽

Vol 102 (3) ◽

pp. 849-857 ◽

Cited By ~ 97

Author(s):

Alexander Röth ◽

Hans Yssel ◽

Jérôme Pène ◽

Elizabeth A. Chavez ◽

Mike Schertzer ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Ectopic Expression ◽

Dominant Negative ◽

Major Influence ◽

Telomeric Dna ◽

Human Telomerase Reverse Transcriptase ◽

Human T Lymphocytes ◽

Human T Cells

Abstract The loss of telomeric DNA with each cell division contributes to the limited replicative lifespan of human T lymphocytes. Although telomerase is transiently expressed in T lymphocytes upon activation, it is insufficient to confer immortality. We have previously shown that immortalization of human CD8+ T lymphocytes can be achieved by ectopic expression of the human telomerase reverse transcriptase (hTERT) gene, which encodes for the catalytic component of the telomerase complex. To study the role of endogenous hTERT in the lifespan of human T cells, we blocked endogenous hTERT expression by ectopic expression of dominant-negative (DN) hTERT. Cells expressing DN-hTERT had a decreased lifespan and showed cytogenetic abnormalities, including chromosome ends without detectable telomeric DNA as well as chromosome fusions. These results indicate that while endogenous hTERT cannot prevent overall telomere shortening, it has a major influence on the longevity of human T cells. Furthermore, we show that up-regulation of hTERT in T cells upon activation decreases over time in culture. Long-term–cultured T cells also show a decreased expression of c-myc upon activation, resulting in less c-myc–induced transcription of hTERT. Moreover, memory T cells, which have expanded in vivo upon antigen encounter, expressed a lower level of hTERT upon activation than naive cells from the same donor. The observed inverse correlation between telomerase levels and replicative history suggests that telomerase levels in T cells are limiting and increasingly insufficient to sustain their proliferation.

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