Expression and activity of P-glycoprotein, a multidrug efflux pump, in human hematopoietic stem cells

Chaudhary, Preet M.; Roninson, Igor B.

doi:10.1016/0092-8674(91)90141-k

Cited by 889 publications

(495 citation statements)

References 24 publications

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“…No inhibition of clonogenicity was found at concentrations of SYTO16 up to 10 nm. (Chaudhary and Roninson, 1991).…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

“…An alternative approach to study the impact of P-gp expression is to measure the P-gp-mediated drug efflux in tumour cells. Such functional P-gp assays have been extensively used to assess drug transporter activity in cell lines and more recently in normal haematopoietic cells (Neyfakh et al, 1989;Chaudhary and Roninson, 1991) and haematological malignancies (Marie et al, 1993;Ross et al, 1993;Ino et al, 1994;Leith et al, 1995). These assays are based on the ability of P-gp to export fluorescent dyes, such as rhodamine 123, out of cells, allowing a convenient flow cytometric determination of P-gp activity.…”

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confidence: 99%

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Highly sensitive and specific detection of P-glycoprotein function for haematological and solid tumour cells using a novel nucleic acid stain

Broxterman

Schuurhuis²,

Lankelma³

et al. 1997

Br J Cancer

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Summary Progress in our understanding of the contribution of P-glycoprotein (P-gp)-mediated resistance to chemotherapy failure in haematological as well as solid tumours has been hampered by the lack of highly sensitive, reliable methods for the detection of P-gp function in fresh human tumour cells. The present study identifies the novel nucleic acid stain SYT016 as a highly sensitive and specific dye to assess P-gp function. The effect of P-gp is expressed here as the ratio of dye fluorescence (RF) from cells incubated with dye with or without 2 gM of the P-gp inhibitor PSC 833. Using flow cytometric analysis, an RF of 0.9 was found for SYT016 in the KB3-1 (P-gp-) and 1.6 in KB8 (P-gp+) cells. Three types of patients' cells were studied: (1) in haematopoietic CD34+ cells, which are known to express P-gp, the RF was 6.0 for SYTO16 compared with 2.5 for rhodamine 123 and 1.3 for daunorubicin (mean of five individuals); (2) in acute myeloid leukaemia cells, the RF for SYTO16 was 1.0 in P-gp-and 4.5 in P-gp+ samples; (3) for the first time, we have quantitated P-gp function in fresh human solid tumour (sarcoma) cells. We found, in a P-gp+ leiomyosarcoma, an RF of 16 for SYT016 and 2.7 for daunorubicin. This means that complete inhibition of P-gp function in these sarcoma cells would lead to an increase of daunorubicin accumulation with 170% compared with 30% in the CD34+ cells. Next, we showed that SYTO16 could be fixed in nuclei by 3.6% formaldehyde treatment, allowing quantification of the nuclear fluorescence on cytospins by laser scanning microscopy. In conclusion, SYTO16 proved to have a combination of favourable properties: it can be excited at 488 nm and has large fluorescence enhancement upon binding to nucleic acids, allowing the use of low, nontoxic (< 10 nM) concentrations. Because the RF for SYT016 is much higher than for daunorubicin, it can be applied for the determination of P-gp function in relatively small numbers of low-P-gp-expressing tumour cells by laser scanning microscopy. Individual sarcomas were found to have high P-gp function compared with CD34+ cells. This assay may be used to select patients for P-gp modulation protocols.

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“…No inhibition of clonogenicity was found at concentrations of SYTO16 up to 10 nm. (Chaudhary and Roninson, 1991).…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

mentioning

confidence: 99%

Highly sensitive and specific detection of P-glycoprotein function for haematological and solid tumour cells using a novel nucleic acid stain

Broxterman

Schuurhuis²,

Lankelma³

et al. 1997

Br J Cancer

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“…56 However, recent works evidenced that MDR1 expression remains limited to naive (CD27 neg ) B cells and undeniably discriminates them from transitional and memory B cells. 57 Moreover, the double MDR1/CD27 staining allows to delineate the switched and transitional B cells, the latter being CD27 neg MDR1 neg .…”

Section: Introductionmentioning

confidence: 99%

“…Pioneer studies on MDR1 in T lymphocytes have shown its expression on only half of CD4 + T cells. 56 In-depth MDR1 functional analysis based on CD45RA/CCR7 expression with Mitotracker green (MTG) dye, 59 showed that naive cells (CD45RA + CCR7 + ) exclude less MTG from their cytoplasm than CD4 + effector (CD45RA neg CCR7 neg , T EM ) and central (CD45RA neg CCR7 +, T CM ) memory cells. 60 The majority of CD4 + T cells expressing MDR1 are therefore memory cells with the exception of Treg (FoxP3 + or CD25 high CD127 low/neg ) lacking MDR1 expression.…”

Section: Introductionmentioning

confidence: 99%

MDR1 in immunity: friend or foe?

et al. 2018

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MDR1 is an ATP-dependent transmembrane transporter primarily studied for its role in the detoxification of tissues and for its implication in resistance of tumor cells to chemotherapy treatment. Several studies also report on its expression on immune cells where it plays a protective role from xenobiotics and toxins. This review provides an overview of what is known on MDR1 expression in immune cells in human, and its implications in different pathologies and their treatment options.

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“…Existing animal models will facilitate the answer to these questions. 24,25 Hacein-Bey et al 26 showed a steady increase in the number of ␥c expressing cells over a period of 6 months in transduced unselected XSCID B cells. However, these results suggesting a selective advantage to cells expressing ␥c might be affected by contaminating producer cells used in the coculture method of transduction.…”

Section: Figure 6 Expression Of Pgp In Human Xscid B Cells Cell Extrmentioning

confidence: 99%

Characterization of an MDR1 retroviral bicistronic vector for correction of X-linked severe combined immunodeficiency

et al. 1998

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Expression and activity of P-glycoprotein, a multidrug efflux pump, in human hematopoietic stem cells

Cited by 889 publications

References 24 publications

Highly sensitive and specific detection of P-glycoprotein function for haematological and solid tumour cells using a novel nucleic acid stain

Highly sensitive and specific detection of P-glycoprotein function for haematological and solid tumour cells using a novel nucleic acid stain

MDR1 in immunity: friend or foe?

Characterization of an MDR1 retroviral bicistronic vector for correction of X-linked severe combined immunodeficiency

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