2000
DOI: 10.4049/jimmunol.164.8.4018
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Engagement of CD28 Modulates CXC Chemokine Receptor 4 Surface Expression in Both Resting and CD3-Stimulated CD4+ T Cells

Abstract: Optimal CD4+ T cell activation requires the cooperation of multiple signaling pathways coupled to the TCR-CD3 complex and to the CD28 costimulatory molecule. In this study, we have investigated the expression of surface CXC chemokine receptor 4 (CXCR4) in enriched populations of CD4+ T PBL, stimulated with anti-CD3 and anti-CD28 mAbs, immobilized on plastic. Anti-CD3 alone induced a progressive down-regulation of surface CXCR4, accompanied by a significant decline in the entry of the HXB2 T cell line-tropic (X… Show more

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Cited by 39 publications
(55 citation statements)
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“…After coating, plates were rinsed with AIM-V serum-free medium (GIBCO͞BRL) to remove nonadherent proteins, and medium was immediately added to the plates after the final wash. It should be noted that the dose of proteins or mAbs bound to each well has been estimated to be approximately 5% of the dose added (6,9), therefore, it is 50 ng per well of anti-CD3 and anti-CD28 mAbs, 20 ng per well of Tat, or equimolar concentrations of each short peptide. Plates were set up in triplicate for all of the experiments described.…”
Section: Methodsmentioning
confidence: 99%
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“…After coating, plates were rinsed with AIM-V serum-free medium (GIBCO͞BRL) to remove nonadherent proteins, and medium was immediately added to the plates after the final wash. It should be noted that the dose of proteins or mAbs bound to each well has been estimated to be approximately 5% of the dose added (6,9), therefore, it is 50 ng per well of anti-CD3 and anti-CD28 mAbs, 20 ng per well of Tat, or equimolar concentrations of each short peptide. Plates were set up in triplicate for all of the experiments described.…”
Section: Methodsmentioning
confidence: 99%
“…Peripheral blood mononuclear cells were isolated by FicollHypaque density-gradient centrifugation (Amersham Pharmacia) of heparinized leukocyte units obtained from 20 HIV-1-seronegative adult donors. Enriched populations of resting CD4 ϩ T cells were isolated by immunomagnetic negative selection with Dynabeads M450 (Dynal, Great Neck, NY; Polysciences) as described (9). We used a mixture of mAbs against CD19, CD20, CD16, CD56, CD57, CD14, and CD8 (Coulter).…”
Section: Methodsmentioning
confidence: 99%
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“…Apoptosis contributes to the massive depletion of CD4 ϩ T-cells and consequently to the loss of immune competence during HIV-1 infection (43,44). Other effects of extracellular Tat include inhibition of the proliferation of uninfected T-cells (24,37,45), possibly by repression of major histocompatibility complex class I transcription (46); regulation of the expression of the HIV-1 coreceptor CXC chemokine receptor-4 on T-lymphocytes (47); and repression of the expression of manganese-superoxide dismutase (48).…”
mentioning
confidence: 99%
“…Naive CD4 + Th cells were isolated from human PBMC by using the CD4 + T Cell Isolation Kit II (Miltenyi Biotec) and the VarioMACS cell Separator, as previously described (27)(28)(29)(30). Purity of cells was immediately checked by anti-CD4-FITC and anti-CD8-PE staining and flow cytometry analysis.…”
Section: Cell Isolationmentioning
confidence: 99%