Maintenance of Normal T Lymphocyte Function after Transfection with SV40 Large T

Ryan, Qin; Goonewardene, I. Michael; Murasko, Donna M.

doi:10.1006/cimm.1993.1136

Cited by 6 publications

(7 citation statements)

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“…However, in contrast to our cell line, the latter two types of transformation give rise to cell lines that are largely dependent on exogenously added IL-2, although they can acquire IL-2 independency after prolonged growth in vitro [2]. The SV40transformed cell line described by Ryan and co-workers is also dependent on IL-2 [7,8].…”

Section: Discussioncontrasting

confidence: 62%

“…With the exception of human leukaemic or lymphoma cell lines, there are a few examples of transformed T-lymphocyte cell lines. They have been obtained by transfection with EBV DNA [6], transformation with HTLV-1 [2,3] or Herpesvirus saimiri [2,4,5], or recently by transfection with SV40 large T [7,8]. Human, EBV-infected transformed T-cell lines have also been described, which were isolated from patients [38,39] or very recently from established EBV-transformed lymphoblastoid-cell lines [40].…”

Section: Discussionmentioning

confidence: 99%

“…In order to circumvent this problem, cells obtained from human lymphoblastic leukaemias or lymphomas are often used because of their unlimited lifespan. Transformation of mature human T cells has also been attempted using various approaches, such as infection with human T-cell leukaemia virus-type 1 (HTLV-1) [2,3] or Herpesvirus saimiri [4,5] and transfection with EBV [6] or simian virus 40 (SV40) DNA [7,8]. Although they are continuously growing, all these transformed human T-cell lines usually remain dependent on exogenously added IL-2 [2].…”

Section: Introductionmentioning

confidence: 99%

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Establishment and Characterization of a Human T‐Lymphocyte Cell Line Immortalized by SV40 and with Abnormal Expression of TCR/CD3

et al. 1998

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Human lymphocytes derived from the peripheral blood of a healthy woman were transfected with a plasmid carrying the simian virus 40 (SV40) large T antigen. The successfully transformed cells contained SV40 large T DNA and were negative for Epstein-Barr virus (EBV) and human T-cell leukaemia virus (HTLV)-1 genomes. The immortalized cell line was assigned to the T-lymphocyte lineage on the basis of morphological, immunological and cytochemical criteria. While the cells expressed CD1a and CD4 at the cell surface, the CD3 complex was solely intracytoplasmic. Immunoprecipitation studies indicated that these cells lacked T-cell receptor (TCR) alpha-chains but not beta-chains. They were negative for activation markers such as CD25, CD69 and major histocompatibility (MHC) class II molecules. In addition, the transformed cells exhibited a complete growth independency towards interleukin-2 (IL-2). However, after phorbol ester stimulation, CD25 and CD69 markers were expressed and IL-2 was secreted. This new human immortalized T-lymphocytic cell line, which is cell-surface TCR/CD3-negative, may be useful as an in vitro model for studying TCR/CD3 assembly, expression and signal transduction.

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Section: Discussioncontrasting

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Establishment and Characterization of a Human T‐Lymphocyte Cell Line Immortalized by SV40 and with Abnormal Expression of TCR/CD3

et al. 1998

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“…The human T-cell leukaemia virus type-1 (HTLV-1) oncoprotein, Tax, which is crucial for transformation by HTLV-1, has recently been shown to bind and inactivate p16 by interacting with its ankyrin repeats, indicating a mechanism for the transforming abilities of this virus (Suzuki et al, 1996). Furthermore, transfection of primary T-lymphocytes with SV40 large T, whose protein is known to abrogate the pRb pathway, delays the point of cellular senescence (Ryan et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

Involvement of the Ink4 proteins p16 and p15 in T-lymphocyte senescence

Erickson¹,

Sangfelt²,

Heyman³

et al. 1998

Oncogene

104

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Little is known about the molecular background to senescence in T-lymphocytes. In ®broblast systems replicative senescence has been shown to correlate with a number of changes in the expression of the proteins normally regulating progression through the G1 phase of the cell cycle, and recently the Ink4 inhibitor p16 was implicated as a central regulator of replicative senescence in human ®broblasts. It has, however, been claimed that p16 is not expressed in T-lymphocytes. In the present study we have analysed G1 regulating proteins in ageing human T-lymphocytes. We show that PHA and IL-2 stimulated T-lymphocytes cease to proliferate after around 20 population doublings, these cells can not thereafter be restimulated to growth, and were also found to exhibit markers for senescence. We found that Tlymphocytes accumulate p16 and p15 protein during successive population doublings and display high levels of these proteins as they enter into replicative senescence. There was also an increased binding of p16 to the Cdk6 kinase in senescent cells, and a decreased Cdk6 as well as Cdk2 kinase activity. The levels of other G1 regulating proteins were also altered in the senescent cells, such as slightly elevated levels of p21/WAF1, and downregulation of Cdk2 and cyclinD3. The levels of p27/ Kip1 is down regulated in proliferating cells but rise to approximately 15% of the levels in un-stimulated quiescent cells. As a high proportion of T-cell childhood acute lymphoblastic leukaemias have deletions of both p15 and p16, our data suggest that inactivation of these genes makes it possible for leukemic cells to avoid senescence.

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“…CXCR6 + human peripheral lymphocytes were isolated from buffy coats of five healthy human volunteers by Ficoll separation followed by CXCR6 staining with the monoclonal anti‐human CXCR6‐PE antibody (2.5μxg/∼1 × 10 6 cells), sorting using the FACS Aria (Becton Dickinson and propagated with human recombinant Interleukin‐2 (25 U/ml) as reported previously [28].…”

Section: Methodsmentioning

confidence: 99%

Homocysteine up‐regulates vascular transmembrane chemokine CXCL16 and induces CXCR6+ lymphocyte recruitment in vitro and in vivo

Postea

Koenen

Hristov

et al. 2008

J Cellular Molecular Medi

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Objective: Hyperhomocysteinemia induces endothelial dysfunction and promotes atherosclerotic vascular disease. Infiltrates of activated macrophages and lymphocytes are observed in human and experimental atherosclerotic lesions, their emigration being guided by endothelial-leukocyte adhesion molecules and chemoattractants. The CXC-chemokine CXCL16 functions as an adhesion molecule by interacting with its receptor (CXCR6) and also as a scavenger for oxidized low density lipoprotein (oxLDL). We investigated the modulation of CXCL16 on cultured endothelial cells (EC) and the recruitment of CXCR6+ lymphocytes in response to homocysteine (Hcy), in vitro and in vivo. Methods and Results: Hcy-stimulated EC show a significant increase in CXCL16 mRNA and protein expression. Incubation of EC with d,l-Hcy and l-Hcy significantly increased CXCR6+ lymphocyte adhesion to EC while l-Cysteine (l-Cys) had no effect. Furthermore, EC stimulation with Hcy increased uptake of DiI-oxLDL. An anti-CXCL16 monoclonal antibody, antioxidants (Tiron) and PPAR-γ agonists (Pioglitazone) considerably reduced CXCR6+ lymphocyte adhesion and uptake of DiI-oxLDL. Upon injection in the peritoneal cavities of mice, l-Hcy and not l-Cys, increased the number of CXCR6+ lymphocytes, which was reduced by coinjection with Pioglitazone or anti-human CXCL16 antibody. Conclusions: Hyperhomocysteinemia up-regulates CXCL16 leading to increased recruitment of CXCR6+ lymphocytes and scavenging of modified lipids via a potential involvement of a PPAR-γ-dependent mechanism. CXCL16 may therefore contribute to the formation and progression of atherosclerotic lesions under conditions of hyperhomocysteinemia.

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Maintenance of Normal T Lymphocyte Function after Transfection with SV40 Large T

Cited by 6 publications

References 0 publications

Establishment and Characterization of a Human T‐Lymphocyte Cell Line Immortalized by SV40 and with Abnormal Expression of TCR/CD3

Establishment and Characterization of a Human T‐Lymphocyte Cell Line Immortalized by SV40 and with Abnormal Expression of TCR/CD3

Involvement of the Ink4 proteins p16 and p15 in T-lymphocyte senescence

Homocysteine up‐regulates vascular transmembrane chemokine CXCL16 and induces CXCR6+ lymphocyte recruitment in vitro and in vivo

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