Ex vivo isolation protocols differentially affect the phenotype of human CD4+ T cells

Bernard, Frédéric; Jaleco, Sara; Dardalhon, Valérie; Steinberg, Marcos; Yssel, Hans; Noraz, Nelly; Taylor, Naomi; Kinet, Sandrina

doi:10.1016/s0022-1759(02)00412-x

Cited by 18 publications

(12 citation statements)

References 26 publications

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“…Under these conditions, the cytokine milieu is more strictly controlled but the effects are clearly biased by the culture conditions as well as the methods used to isolate the selected populations (47,48). In our study, naive and memory CD4 ϩ T cell populations were isolated by negative selection on the basis of CD45RA and CD45RO expression, respectively.…”

Section: Discussionmentioning

confidence: 99%

Homeostasis of Naive and Memory CD4+ T Cells: IL-2 and IL-7 Differentially Regulate the Balance Between Proliferation and Fas-Mediated Apoptosis

Jaleco

Swainson

Dardalhon

et al. 2003

The Journal of Immunology

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Cytokines play a crucial role in the maintenance of polyclonal naive and memory T cell populations. It has previously been shown that ex vivo, the IL-7 cytokine induces the proliferation of naive recent thymic emigrants (RTE) isolated from umbilical cord blood but not mature adult-derived naive and memory human CD4+ T cells. We find that the combination of IL-2 and IL-7 strongly promotes the proliferation of RTE, whereas adult CD4+ T cells remain relatively unresponsive. Immunological activity is controlled by a balance between proliferation and apoptotic cell death. However, the relative contributions of IL-2 and IL-7 in regulating these processes in the absence of MHC/peptide signals are not known. Following exposure to either IL-2 or IL-7 alone, RTE, as well as mature naive and memory CD4+ T cells, are rendered only minimally sensitive to Fas-mediated cell death. However, in the presence of the two cytokines, Fas engagement results in a high level of caspase-dependent apoptosis in both RTE as well as naive adult CD4+ T cells. In contrast, equivalently treated memory CD4+ T cells are significantly less sensitive to Fas-induced cell death. The increased susceptibility of RTE and naive CD4+ T cells to Fas-induced apoptosis correlates with a significantly higher IL-2/IL-7-induced Fas expression on these T cell subsets than on memory CD4+ T cells. Thus, IL-2 and IL-7 regulate homeostasis by modulating the equilibrium between proliferation and apoptotic cell death in RTE and mature naive and memory T cell subsets.

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Section: Discussionmentioning

confidence: 99%

Homeostasis of Naive and Memory CD4+ T Cells: IL-2 and IL-7 Differentially Regulate the Balance Between Proliferation and Fas-Mediated Apoptosis

Jaleco

Swainson

Dardalhon

et al. 2003

The Journal of Immunology

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“…Moreover, when these authors assessed GLUT1 expression in lymphocytes isolated via negative selection and in the absence of elutriation, only a very low percentage of T cells bound the HTLV SU immunoadhesin (91). In this regard, it is important to note that several recent studies, including work performed in our own laboratory, have shown that the choice of blood coagulant as well as the method used to isolate T lymphocytes can modulate surface expression of receptors pertinent to T lymphocyte function (109).…”

Section: Lymphocyte Activation and Glut1-dependent Metabolismmentioning

confidence: 99%

HTLV envelopes and their receptor GLUT1, the ubiquitous glucose transporter: a new vision on HTLV infection?

Manel

Taylor²,

Kinet³

et al. 2004

Front Biosci

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We identified the ubiquitous glucose transporter GLUT1 as a receptor for Deltaretroviruses HTLV-1 and HTLV-2 envelopes (Env), mediating viral binding and entry. Here, we review the context and key observations that led us to this finding: functional modules of HTLV SU are similar to those of Gammaretrovirus Env which use multimembrane-spanning nutrient transporters as receptors; the HTLV Env receptor is an early marker of T lymphocyte activation; and HTLV Env inhibits glucose transport. We review several molecular, viral, cellular and physiological aspects of HTLV infection in relation to the in vivo and in vitro properties of GLUT1. Also, we examine the implications of HTLV-1 Env-GLUT1 interactions and altered glucose transport on the two major HTLV-1-induced diseases, adult T cell leukemia (ATL) and neurodegenerative tropical spastic paraparesis/HTLV-associated myelopathy (TSP/HAM). Complementary to the classical models of disease progression, we propose new schemes that emphasize the potential metabolic alterations caused in different cellular compartments. Finally, we review the potential use of HTLV Env-derived constructs as tools for labeling GLUT1 in vivo and inhibiting GLUT1 transport in tumor cells.

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“…Although this approach achieves a high grade of purity in the specific cell population (21), several studies have reported that intracellular signaling could be activated downstream from the antibody binding site. The function of positively selected cells could therefore be altered by the procedure used to purify them (22,23). We used anti-vWF antibody for the isolation of ECs and anti-CD 90 for fibroblast isolation.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Culture of Fibroblasts, Vascular Smooth Muscle, and Endothelial Cells From the Fetal Rat Ductus Arteriosus

et al. 2011

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ABSTRACT:The ductus arteriosus (DA), a fetal arterial shunt vessel between the proximal descending aorta and the pulmonary artery, closes shortly after birth. Initial functional closure as a result of the DA's smooth muscle contraction is followed by definite anatomical closure. The latter involves several complex mechanisms like endothelial cushion formation and smooth muscle cell migration resulting in fibrosis and sealing of the vessel. These complex steps indicate highly specialized functions of the DA vascular smooth muscle cells (VSMCs), endothelial cells, and fibroblasts. Herein, we describe a new reproducible method for isolating VSMCs, endothelial cells, and fibroblasts of high viability from fetal rat DA using immunomagnetic cell sorting. Purity of the different cell cultures was assessed by immunohistochemistry and flow cytometry and ranged between 85 and 94%. The capability of the VSMCs to react to hypoxic stimuli was assessed by intracellular calcium and ATP measurements and by VEGF mRNA expression analysis. VSMCs respond to hypoxia with decreases in intracellular calcium concentrations and ATP levels, whereas VEGF mRNA expression increased 3.2-fold. The purified vessel-specific different cell types are suitable for subsequent gene expression profiling and functional studies and provide important tools for improving our understanding of the complex processes involved in the closure of the DA. T he ductus arteriosus (DA) is a shunt which connects the proximal descending aorta and the main pulmonary artery (PA). During fetal life, it serves to bypass the pulmonary circulation. The DA closes shortly after birth, and pulmonary blood flow increases during late gestation (1). After birth, an abrupt increase in oxygen tension results in ductal constriction, which is further promoted by falling levels of prostaglandin E2 because of an increased pulmonary metabolism and the elimination of placental prostaglandin. DA medial smooth muscle contraction leads to wall thickening, luminal obliteration, and shortening of the vessel. Permanent sealing of the DA is a process generated by the infolding of the endothelium, neointima formation, subintimal disruption, and vasa vasorum ingrowth resulting in subsequent vessel fibrosis. Several major molecular pathways are known to be involved in this programmed proliferation during permanent closure of the DA, e.g. NO signaling, the system of VEGF-promoted angiogenesis, and the cyclooxygenase/prostaglandin system (2-5).However, many questions concerning the exact function and cross-talk of these signaling pathways and the molecular basis for failed DA closure still remain unanswered (6). Previous studies investigating molecular events in the duct were in part hampered by the lack of an in vitro model that allows to study the processes in question separately in each of the different cell types of the DA instead of using the whole vessel (7). Herein, we describe an improved and reproducible method to culture organotypic vascular smooth muscle cells (VSMCs), fibroblasts, and e...

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Ex vivo isolation protocols differentially affect the phenotype of human CD4+ T cells

Cited by 18 publications

References 26 publications

Homeostasis of Naive and Memory CD4+ T Cells: IL-2 and IL-7 Differentially Regulate the Balance Between Proliferation and Fas-Mediated Apoptosis

Homeostasis of Naive and Memory CD4+ T Cells: IL-2 and IL-7 Differentially Regulate the Balance Between Proliferation and Fas-Mediated Apoptosis

HTLV envelopes and their receptor GLUT1, the ubiquitous glucose transporter: a new vision on HTLV infection?

Isolation and Culture of Fibroblasts, Vascular Smooth Muscle, and Endothelial Cells From the Fetal Rat Ductus Arteriosus

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