scholarly journals P-glycoprotein expression and function in circulating blood cells from normal volunteers

Blood ◽  
1994 ◽  
Vol 83 (9) ◽  
pp. 2451-2458 ◽  
Author(s):  
WT Klimecki ◽  
BW Futscher ◽  
TM Grogan ◽  
WS Dalton
Keyword(s):  
Nk Cell ◽  
Efflux Pump ◽  
Immunoblot Analysis ◽  
Rhodamine 123 ◽  
Double Labeling ◽  
Normal Volunteers ◽  
P Glycoprotein ◽  
Mdr1 Mrna ◽  
Mrna Analysis ◽  
And Function

Abstract In contrast to its clearly defined role as a multidrug efflux pump in neoplastic cells, the physiologic function of P-glycoprotein (P-gly) in normal cells is unclear. Recent reports identifying P-gly in normal blood and bone marrow suggest that hematopoietic development or function may be dependent on P-gly. To understand the normal function of P-gly in the blood, its level of expression and function must first be quantitated relative to a known standard. In this study, P-gly, MDR1 gene expression, and P-gly function were quantitated in normal leukocytes. P-gly and MDR1 expression were analyzed in individual leukocyte lineages (T-helper, T-suppressor, monocyte, granulocyte, B- lymphocyte, NK cell) from normal volunteers. P-gly on the cell surface was detected by fluorescent double-labeling for lineage (CD4, CD8, CD14, CD15, CD19, CD56, respectively) and P-gly (MRK16) with analysis by flow cytometry and in some cases immunoblot analysis. MDR1 mRNA analysis on purified lineages was performed using quantitative reverse transcription-polymerase chain reaction. P-gly function was determined for each lineage using dual-labeling for lineage and P-gly substrate (rhodamine 123). The P-gly expressing human myeloma cell line, 8226/Dox6, was used as a reference of comparison for levels of P-gly, MDR1 mRNA, and function. CD56+ cells expressed the highest levels of MDR1 mRNA followed by CD8+ > CD4+ approximately equal to CD15+ > CD19+ > CD14+, with percentage values relative to Dox6 of 49%, 17%, 8%, 8%, 4%, and 2%, respectively. The assays for P-gly immunofluorescence and function correlated well with mRNA analysis except for CD15+ cells (granulocytes), which showed a moderate MDR1 mRNA level with a lack of both function and surface P-gly staining. Granulocyte membranes did show P-gly on immunoblot analysis when probed with either C219 or JSB1. We conclude that (1) P-gly and the MDR1 mRNA are expressed in normal leukocytes, (2) this P-gly expression is lineage specific with relatively high levels among CD56+ cells, and (3) the expression of P- gly in granulocytes is not associated with transport of the P-gly substrate, rhodamine 123, out of the cell.

scholarly journals P-glycoprotein expression and function in circulating blood cells from normal volunteers

Blood ◽  
1994 ◽  
Vol 83 (9) ◽  
pp. 2451-2458 ◽  
Author(s):  
WT Klimecki ◽  
BW Futscher ◽  
TM Grogan ◽  
WS Dalton
Keyword(s):  
Nk Cell ◽  
Efflux Pump ◽  
Immunoblot Analysis ◽  
Rhodamine 123 ◽  
Double Labeling ◽  
Normal Volunteers ◽  
P Glycoprotein ◽  
Mdr1 Mrna ◽  
Mrna Analysis ◽  
And Function

In contrast to its clearly defined role as a multidrug efflux pump in neoplastic cells, the physiologic function of P-glycoprotein (P-gly) in normal cells is unclear. Recent reports identifying P-gly in normal blood and bone marrow suggest that hematopoietic development or function may be dependent on P-gly. To understand the normal function of P-gly in the blood, its level of expression and function must first be quantitated relative to a known standard. In this study, P-gly, MDR1 gene expression, and P-gly function were quantitated in normal leukocytes. P-gly and MDR1 expression were analyzed in individual leukocyte lineages (T-helper, T-suppressor, monocyte, granulocyte, B- lymphocyte, NK cell) from normal volunteers. P-gly on the cell surface was detected by fluorescent double-labeling for lineage (CD4, CD8, CD14, CD15, CD19, CD56, respectively) and P-gly (MRK16) with analysis by flow cytometry and in some cases immunoblot analysis. MDR1 mRNA analysis on purified lineages was performed using quantitative reverse transcription-polymerase chain reaction. P-gly function was determined for each lineage using dual-labeling for lineage and P-gly substrate (rhodamine 123). The P-gly expressing human myeloma cell line, 8226/Dox6, was used as a reference of comparison for levels of P-gly, MDR1 mRNA, and function. CD56+ cells expressed the highest levels of MDR1 mRNA followed by CD8+ > CD4+ approximately equal to CD15+ > CD19+ > CD14+, with percentage values relative to Dox6 of 49%, 17%, 8%, 8%, 4%, and 2%, respectively. The assays for P-gly immunofluorescence and function correlated well with mRNA analysis except for CD15+ cells (granulocytes), which showed a moderate MDR1 mRNA level with a lack of both function and surface P-gly staining. Granulocyte membranes did show P-gly on immunoblot analysis when probed with either C219 or JSB1. We conclude that (1) P-gly and the MDR1 mRNA are expressed in normal leukocytes, (2) this P-gly expression is lineage specific with relatively high levels among CD56+ cells, and (3) the expression of P- gly in granulocytes is not associated with transport of the P-gly substrate, rhodamine 123, out of the cell.

Expression of Functionally P-Glycoprotein in MA104 Kidney Cells

1999 ◽  
Vol 54 (1-2) ◽  
pp. 119-127 ◽  
Author(s):  
Márcia A. M. Capella ◽  
Marcelo Marcos Morales ◽  
Monique Orind ◽  
Vivian M. Rumjanek ◽  
Aníbal Gil Lopes
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleotide Sequence ◽  
High Expression ◽  
Kidney Cells ◽  
Chain Reaction ◽  
High Identity ◽  
P Glycoprotein ◽  
Mdr1 Mrna ◽  
Polymerase Chain ◽  
And Function

Rhesus monkey kidney MA104 cells are a polarized epithelium with some unusual characteristics, including a resistance to ouabain, although their Na+-K+-ATPase has normal affinity with this drug. This work suggests that MA104 cells have high expression of functionally P-glycoprotein in their membranes. This was established using four complementary methods to investigate the expression and function of P-glycoprotein in these cells. MA104 cells were strongly resistant to vincristine, which could be reversed by three know n P-glycoprotein modulators; verapamil, cyclosporin A and trifluoperazine. In addition, MA104 cells accumulate little rhodamine 123, and the incubation with verapamil increased this accumulation. The mdr1-mRNA was detected by reverse transcription-polymerase chain reaction and a subcloned 283-bp product was identified. Its nucleotide sequence was compared with the related region of hum an mdr1, showing a high identity (96% ) between the two sequences. The expression of P-glycoprotein in the cell membrane was observed by Western blot and immunofluorescence. The results taken together suggest that MA104 cells intrinsically have a high expression of functionally P-glycoprotein in their membranes

P-glycoprotein efflux pump expression and activity in Calu-3 cells

2001 ◽  
Vol 90 (5) ◽  
pp. 645
Author(s):  
Karen O. Hamilton ◽  
Gunilla Backstrom ◽  
Mehran A. Yazdanian ◽  
Kenneth L. Audus
Keyword(s):  
Efflux Pump ◽  
P Glycoprotein ◽  
Expression And Activity

scholarly journals Sex- and age‐dependent alterations of splenic immune cell profile and NK cell phenotypes and function in C57BL/6J mice

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Kelly B. Menees ◽  
Rachael H. Earls ◽  
Jaegwon Chung ◽  
Janna Jernigan ◽  
Nikolay M. Filipov ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Sex Differences ◽  
Nk Cells ◽  
Immune Cell ◽  
Nk Cell ◽  
Spleen Weight ◽  
Age Related ◽  
And Function ◽  
Cell Phenotypes

Abstract Background Physiological homeostasis decline, immunosenescence, and increased risk for multiple diseases, including neurodegeneration, are all hallmarks of ageing. Importantly, it is known that the ageing process is sex-biased. For example, there are sex differences in predisposition for multiple age-related diseases, including neurodegenerative and autoimmune diseases. However, sex differences in age-associated immune phenotypes are not clearly understood. Results Here, we examined the effects of age on immune cell phenotypes in both sexes of C57BL/6J mice with a particular focus on NK cells. We found female-specific spleen weight increases with age and concordant reduction in the number of splenocytes per gram of spleen weight compared to young females. To evaluate sex- and age-associated changes in splenic immune cell composition, we performed flow cytometry analysis. In male mice, we observed an age-associated reduction in the frequencies of monocytes and NK cells; female mice displayed a reduction in B cells, NK cells, and CD8 + T cells and increased frequency of monocytes and neutrophils with age. We then performed a whole blood stimulation assay and multiplex analyses of plasma cytokines and observed age- and sex-specific differences in immune cell reactivity and basal circulating cytokine concentrations. As we have previously illustrated a potential role of NK cells in Parkinson’s disease, an age-related neurodegenerative disease, we further analyzed age-associated changes in NK cell phenotypes and function. There were distinct differences between the sexes in age-associated changes in the expression of NK cell receptors, IFN-γ production, and impairment of α-synuclein endocytosis. Conclusions This study demonstrates sex- and age-specific alterations in splenic lymphocyte composition, circulating cytokine/chemokine profiles, and NK cell phenotype and effector functions. Our data provide evidence that age-related physiological perturbations differ between the sexes which may help elucidate sex differences in age-related diseases, including neurodegenerative diseases, particularly Parkinson’s disease, where immune dysfunction is implicated in their etiology.

scholarly journals Expression and Function of Organic Anion Transporting Polypeptides in the Human Brain: Physiological and Pharmacological Implications

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 834
Author(s):  
Anima M. Schäfer ◽  
Henriette E. Meyer zu Schwabedissen ◽  
Markus Grube
Keyword(s):  
Organic Anion ◽  
Physiological Role ◽  
Point Of View ◽  
Efflux Transporters ◽  
Organic Anion Transporting Polypeptides ◽  
Current State ◽  
P Glycoprotein ◽  
The Central Nervous System ◽  
And Function ◽  
Uptake Transporters

The central nervous system (CNS) is an important pharmacological target, but it is very effectively protected by the blood–brain barrier (BBB), thereby impairing the efficacy of many potential active compounds as they are unable to cross this barrier. Among others, membranous efflux transporters like P-Glycoprotein are involved in the integrity of this barrier. In addition to these, however, uptake transporters have also been found to selectively uptake certain compounds into the CNS. These transporters are localized in the BBB as well as in neurons or in the choroid plexus. Among them, from a pharmacological point of view, representatives of the organic anion transporting polypeptides (OATPs) are of particular interest, as they mediate the cellular entry of a variety of different pharmaceutical compounds. Thus, OATPs in the BBB potentially offer the possibility of CNS targeting approaches. For these purposes, a profound understanding of the expression and localization of these transporters is crucial. This review therefore summarizes the current state of knowledge of the expression and localization of OATPs in the CNS, gives an overview of their possible physiological role, and outlines their possible pharmacological relevance using selected examples.

Examination of the Functional Activity of P-glycoprotein in the Rat Placental Barrier Using Rhodamine 123

2003 ◽  
Vol 305 (3) ◽  
pp. 1239-1250 ◽  
Author(s):  
Petr Pavek ◽  
Frantisek Staud ◽  
Zdenek Fendrich ◽  
Hana Sklenarova ◽  
Antonin Libra ◽  
...  
Keyword(s):  
Functional Activity ◽  
Rhodamine 123 ◽  
Placental Barrier ◽  
P Glycoprotein

Expression and function of NK cell receptors in CD8+ T cells

2001 ◽  
Vol 13 (4) ◽  
pp. 465-470 ◽  
Author(s):  
Christopher W McMahon ◽  
David H Raulet
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Nk Cell ◽  
Cell Receptors ◽  
Nk Cell Receptors ◽  
And Function

NKR-P1B Receptor Expression and Function in the Different Innate Lymphoid Cells of the Lung

2021 ◽  
Author(s):  
Lucas Vajko
Keyword(s):  
Nk Cell ◽  
Γδ T Cells ◽  
Bronchial Epithelium ◽  
Lung Parenchyma ◽  
Recognition System ◽  
Innate Lymphoid Cells ◽  
Lymphoid Cells ◽  
Receptor Expression ◽  
And Function ◽  
Deficient Mice

Group 2 innate lymphoid cells (ILC2) are the majority of ILCs in murine lungs at steady state. ILC2s are the main producer of type-2-cytokines, IL-4, IL-5, IL-9, IL-13, and amphiregulin, playing key roles in lung tissue homeostasis, airway responses to pathogens and allergens, and in cancer-related defenses. ILC functions are regulated by cell surface receptors. NKR-P1B is an inhibitory receptor, which recognizes C-type lectin-related protein (Clr-b) as its ligand. NKR-P1B is expressed on subsets of natural killer cells, ILC2, ILC3, γδ T cells, macrophages and dendritic cells in a tissue-specific manner and regulates NK cell and ILC3 functions in the gut. Expression and function of NKR-P1B in the lung ILC populations is unexplored. Moreover, Clr-b, the ligand for NKR-P1B, is expressed in the bronchial epithelium, endothelial cells and in lung parenchyma, but its role in immune regulation in the lung is unknown. We hypothesize that ILC2s in the lung express NKR-P1B, and their function is regulated by the NKR-P1B:Clr-b recognition system. Using wild-type (WT) and NKR-P1B-deficient mice, we study the expression of NKR-P1B on lung ILC2, and the function of NKR-P1B:Clr-b recognition system in ILC2 development and function. We compare the phenotype, frequency, numbers and cytokine production by ILC2s upon stimulation between WT and NKR-P1B-deficient mice using antibody staining and flow cytometry analysis. This study will reveal the role of NKR-P1B as a model system for its human homolog, NKR-P1A, in the regulation of ILC development and function, advancing our understanding of how immune responses in the lung are regulated.

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