Complexity of the primary genetic response to mitogenic activation of human T cells

1989 ◽  
Vol 9 (3) ◽  
pp. 1041-1048
Author(s):  
P F Zipfel ◽  
S G Irving ◽  
K Kelly ◽  
U Siebenlist
Keyword(s):  
T Cells ◽  
Cyclosporin A ◽  
Human Peripheral Blood ◽  
Human Fibroblasts ◽  
Primary Response ◽  
Cdna Clones ◽  
Genetic Response ◽  
Activated T Cells ◽  
Isolation And Characterization ◽  
Inducible Genes

We describe the isolation and characterization of more than 60 novel cDNA clones that constitute part of the immediate genetic response to resting human peripheral blood T cells after mitogen activation. This primary response was highly complex, both in the absolute number of inducible genes and in the diversity of regulation. Although most of the genes expressed in activated T cells were shared with the activation response of normal human fibroblasts, a significant number were more restricted in tissue specificity and thus likely encode or effect the differentiated functions of activated T cells. The activatable genes could be further differentiated on the basis of kinetics of induction, response to cycloheximide, and sensitivity to the immunosuppressive drug cyclosporin A. It is of note that cyclosporin A inhibited the expression of more than 10 inducible genes, which suggests that this drug has a broad genetic mechanism of action.

scholarly journals Complexity of the primary genetic response to mitogenic activation of human T cells.

1989 ◽  
Vol 9 (3) ◽  
pp. 1041-1048 ◽  
Author(s):  
P F Zipfel ◽  
S G Irving ◽  
K Kelly ◽  
U Siebenlist
Keyword(s):  
T Cells ◽  
Cyclosporin A ◽  
Human Peripheral Blood ◽  
Human Fibroblasts ◽  
Primary Response ◽  
Cdna Clones ◽  
Genetic Response ◽  
Activated T Cells ◽  
Isolation And Characterization ◽  
Inducible Genes

We describe the isolation and characterization of more than 60 novel cDNA clones that constitute part of the immediate genetic response to resting human peripheral blood T cells after mitogen activation. This primary response was highly complex, both in the absolute number of inducible genes and in the diversity of regulation. Although most of the genes expressed in activated T cells were shared with the activation response of normal human fibroblasts, a significant number were more restricted in tissue specificity and thus likely encode or effect the differentiated functions of activated T cells. The activatable genes could be further differentiated on the basis of kinetics of induction, response to cycloheximide, and sensitivity to the immunosuppressive drug cyclosporin A. It is of note that cyclosporin A inhibited the expression of more than 10 inducible genes, which suggests that this drug has a broad genetic mechanism of action.

Accessory function of interleukin-1 and interleukin-6: preferential costimulation of T4 positive lymphocytes

Blood ◽  
1990 ◽  
Vol 75 (4) ◽  
pp. 922-930 ◽  
Author(s):  
G Tosato ◽  
J Miller ◽  
G Marti ◽  
SE Pike
Keyword(s):  
T Cells ◽  
T Cell ◽  
Interleukin 6 ◽  
Human Peripheral Blood ◽  
Interleukin 1 ◽  
Small Degree ◽  
Activated T Cells ◽  
Accessory Function ◽  
Wide Range ◽  
Marked Preference

Abstract Two monocyte-derived cytokines, interleukin-1 (IL-1) and interleukin-6 (IL-6), have been reported to costimulate monocyte-depleted T cell populations in the presence of mitogen, and this effect has been attributed to an accessory function of these molecules. We have now examined further the accessory function potential of IL-1 plus IL-6, and examined how these cytokines promote T cell growth with mitogen. Together, IL-1 and IL-6 additively and, to a small degree, synergistically promote the proliferation of highly purified human peripheral blood T cells with phytohemagglutinin (PHA). However, maximum costimulation by IL-1 plus IL-6 over a wide range of concentrations is significantly smaller than that induced by optimal numbers of monocytes. Also, in contrast to monocytes that costimulate equally effectively T4 positive and T8 positive cells, IL-1 plus IL-6 costimulate T4 positive lymphocytes in marked preference to T8 positive cells. IL-1 plus IL-6 induces IL-2 secretion in T cell cultures costimulated with PHA, and an antibody to the IL-2 receptor, anti-Tac, markedly inhibits PHA-activated T cells costimulated by IL-1 plus IL-6. In addition, IL-1 plus IL-6 enhances the expression of surface IL-2 receptors. Because the costimulatory effect of IL-1 plus IL-6 is quantitatively smaller than that of monocytes, and it is preferentially directed toward T4 positive as opposed to T8 positive T cells, IL-1 plus IL-6, together, appear to represent a selective set of monocyte- derived accessory signals.

Effect of rapamycin on the cyclosporin A–resistant CD28-mediated costimulatory pathway

Blood ◽  
2002 ◽  
Vol 99 (12) ◽  
pp. 4517-4524 ◽  
Author(s):  
Paritosh Ghosh ◽  
Meredith A. Buchholz ◽  
Shingo Yano ◽  
Dennis Taub ◽  
Dan L. Longo
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cyclosporin A ◽  
T Cell Activation ◽  
Messenger Rna ◽  
Cell Activation ◽  
Human Peripheral Blood ◽  
Interleukin 2 ◽  
Immunosuppressive Drug ◽  
Receptor Interaction

The consequences of T-cell activation depend exclusively on costimulation during antigen–T-cell receptor interaction. Interaction between the T-cell coreceptor CD28 and its ligand B7 during antigen-antigen receptor engagement results in full activation of T cells, the outcomes of which are proliferation and effector functions. The ability of CD28 to costimulate the production of interleukin-2 (IL-2) explains the importance of this costimulation. The signaling event mediated by CD28 engagement has been proposed to have 2 components: one is sensitive to the immunosuppressive drug cyclosporin A (CsA), and the other one is CsA-resistant. In this report, we demonstrate that the CsA-resistant pathway is sensitive to the immunosuppressive drug rapamycin. Treatment with rapamycin blocked IL-2 production after activation of human peripheral blood T cells with phorbol ester (PMA) and anti-CD28 (CsA-resistant pathway), whereas this drug did not have any effect on PMA plus ionomycin stimulation (CsA-sensitive pathway). The inhibitory effect of rapamycin was on messenger RNA stability and translation, rather than on IL-2 transcription or protein turnover.

Effects of Ca-Containing Phosphate Glasses on T Lymphocytes In Vitro

2005 ◽  
Vol 284-286 ◽  
pp. 597-602 ◽  
Author(s):  
A. Kesisoglou ◽  
Jonathan C. Knowles ◽  
I. Olsen
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Dna Synthesis ◽  
T Cell Activation ◽  
Cell Activation ◽  
Human Peripheral Blood ◽  
Suppressor Cells ◽  
Phosphate Glasses ◽  
Activated T Cells

Calcium phosphate-based glasses (PG) are of interest as both scaffold and delivery materials for tissue rebuilding because of their chemical similarity to bone. Since it is essential that these materials exhibit local and systemic biocompatibility and do not adversely affect host tissues, the present study was undertaken to examine the effects of PG containing different amounts of Ca on human T lymphocytes in vitro. This was carried out by measuring the effects of extracts of the PG on the direct and mitogen-induced activation of T cells from human peripheral blood, as well as assessing CD4 and CD8, surface antigens which define T-helper and T-suppressor cells, respectively. The results showed that DNA synthesis by resting T lymphocytes was unaffected by all the PG. However, extracts of the PG containing 24 mol% of Ca caused a very marked inhibition of mitogen-induced T cell activation. This PG also reduced both the resting CD4+ and CD8+ T cells, as well as activated CD8+ cells. In contrast, high Ca-PG significantly augmented DNA synthesis by mitogen-activated T cells. These experiments show that PG containing differing levels of Ca can have pronounced and differential effects on the activation and function of T lymphocytes in vitro.

scholarly journals Regulation of Myosin Heavy Chain Expression during Rat Skeletal Muscle Development In Vitro

2001 ◽  
Vol 12 (5) ◽  
pp. 1499-1508 ◽  
Author(s):  
Carol E. Torgan ◽  
Mathew P. Daniels
Keyword(s):  
Skeletal Muscle ◽  
T Cells ◽  
Cyclosporin A ◽  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Fiber Formation ◽  
Immunofluorescent Staining ◽  
Nuclear Factor ◽  
Activated T Cells

Signals that determine fast- and slow-twitch phenotypes of skeletal muscle fibers are thought to stem from depolarization, with concomitant contraction and activation of calcium-dependent pathways. We examined the roles of contraction and activation of calcineurin (CN) in regulation of slow and fast myosin heavy chain (MHC) protein expression during muscle fiber formation in vitro. Myotubes formed from embryonic day 21 rat myoblasts contracted spontaneously, and ∼10% expressed slow MHC after 12 d in culture, as seen by immunofluorescent staining. Transfection with a constitutively active form of calcineurin (CN*) increased slow MHC by 2.5-fold as determined by Western blot. This effect was attenuated 35% by treatment with tetrodotoxin and 90% by administration of the selective inhibitor of CN, cyclosporin A. Conversely, cyclosporin A alone increased fast MHC by twofold. Cotransfection with VIVIT, a peptide that selectively inhibits calcineurin-induced activation of the nuclear factor of activated T-cells, blocked the effect of CN* on slow MHC by 70% but had no effect on fast MHC. The results suggest that contractile activity-dependent expression of slow MHC is mediated largely through the CN–nuclear factor of activated T-cells pathway, whereas suppression of fast MHC expression may be independent of nuclear factor of activated T-cells.

Immunotherapy with Anti-CD3 x Anti-CMV Bispecific Antibody (CMVBi) Armed Donor Derived Activated T Cells (ATC) – A Novel Strategy against Cytomegalovirus (CMV) Post Allogeneic Stem Cell Transplantation (SCT).

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1157-1157
Author(s):  
Mayur S Ramesh ◽  
Archana Thakur ◽  
Philip Pellett ◽  
Subhendu Das ◽  
Zaid S Al-Kadhimi ◽  
...  
Keyword(s):  
T Cells ◽  
Bispecific Antibody ◽  
Ex Vivo ◽  
Human Fibroblasts ◽  
Interleukin 2 ◽  
Target Cells ◽  
Normal Donor ◽  
Activated T Cells ◽  
Specific Cytotoxicity ◽  
Novel Strategy

Abstract Abstract 1157 Poster Board I-179 Introduction CMV reactivation and infection can cause profound negative outcome post allogeneic SCT. Current management strategies against CMV are suboptimal and are associated with adverse effects. Induction of anti-CMV T cell responses by vaccination has not been helpful in immunocompromised hosts. Immunotherapy with CMV specific donor-derived cytotoxic T lymphocytes (CTL) is effective after allografting, but it is expensive, labor intensive, and diffcult to replicate in most centers. Non-toxic targeted therapy is needed to improve clinical outcomes. In earlier studies, we generated anti-CD3 chemically heteroconjugated with anti-Her2/neu or anti-CD20 bispecific antibody (BiAb). Ex vivo expanded anti-CD3 activated T cells (ATC) and armed with BiAb exhibit high levels of specific cytotoxicity directed at the respective tumor antigens. We propose a novel strategy of using ATC armed ex-vivo with engineered CMVBi to target CMV antigens. We tested the strategy initially in an in vitro cell culture model using CMV-infected fibroblasts and T cells from normal human donors. We hypothesize that donor anti-CD3 ATC armed with CMVBi will target and eliminate CMV-infected target cells. Materials and methods Normal donor peripheral blood mononuclear cells (PBMC) were used to generate ATC by activation with anti-CD3 (OKT3) and interleukin 2 (IL-2). CMVBi was created by chemical heteroconjugation of OKT3 (murine IgG2a) monoclonal antibody and polyclonal anti-CMV (Cytogam®). Specific cytotoxicity was tested by 51Cr release assay using CMV-infected or uninfected human fibroblasts as target cells. Effector populations tested included CMVBi armed ATC and ATC alone; additional controls included CMVBi alone, Cytogam® alone, CMVBi armed, and unarmed PBMC. Cytotoxicity was assessed for CMVBi and an irrelevant BiAb at arming doses ranging from 1 to 500 ng/106 ATC with effector to target ratios (E:T) ranging from 25:1 to 3.125:1. Interferon gamma (IFNγ) EliSpots were used to determine cytokine response after exposing CMV-infected and uninfected fibroblasts to unarmed and CMVBi-armed ATC. Results CMVBi arming with as little as 1 ng/106 ATC was significantly more cytotoxic for target cells than unarmed ATC. There was an incremental increase in cytotoxicity with CMVBi armed ATC as the mutiplicity of infection (MOI) was increased in target cells. At all E:T ratios (25:1, 12.5:1, 6.25:1, 3.125:1 ), ATC armed with a dose of 50 ng/106 CMVBi demonstrated markedly enhanced killing of CMV-infected targets (MOI 1) compared to unarmed ATC (see table). In the uninfected control cells, both unarmed and armed ATC lysis was barely detectable over spontaneous lysis. Immunofluorescent studies showed that CMVBi-armed ATC specifically aggregated around GFP fluorescence-marked CMV-infected fibroblasts, whereas unarmed ATC did not aggregate. Cytokine secretion analyzed by IFNγ EliSpot confirmed the superior cytotoxicity of CMVBi-armed ATC. Conclusion We used polyclonal Cytogam® to make CMVBi-armed ATC that were able to kill target cells expressing various CMV antigens with high specificity. CMVBi-armed PBMC was only minimally cytotoxic in comparision to CMVBi-armed ATC. Hence, infusion of CMVBi alone to patients is unlikely to be as effective as infusion of ATC armed ex-vivo with CMVBi. We demonstrated similar degree of cytotoxicity using different donor ATC including CMV sero-negative donors. This effect is independent of MHC mediated antigen presentation hence, overcomes CMV immune escape. This non-MHC restricted specific killing strategy could be easily adapted for the prevention and/or treatment of CMV infection/disease after allogeneic SCT using donor-derived ATC. Our current use of BiAb-armed ATC for cancer immunotherapy in humans illustrates the feasibility of adoptive immunotherapy with CMVBi-armed ATC in SCT. Disclosures Lum: Transtarget Corporation: Founder.

scholarly journals Mechanisms of Rapid Induction of Interleukin-22 in Activated T Cells and Its Modulation by Cyclosporin A

2011 ◽  
Vol 287 (7) ◽  
pp. 4531-4543 ◽  
Author(s):  
Ina Rudloff ◽  
Malte Bachmann ◽  
Josef Pfeilschifter ◽  
Heiko Mühl
Keyword(s):  
T Cells ◽  
Cyclosporin A ◽  
Activated T Cells ◽  
Interleukin 22 ◽  
Rapid Induction

scholarly journals Cloning and expression of cyclosporin A- and FK506-sensitive nuclear factor of activated T-cells: NF45 and NF90.

1994 ◽  
Vol 269 (32) ◽  
pp. 20691-20699 ◽  
Author(s):  
P.N. Kao ◽  
L. Chen ◽  
G. Brock ◽  
J. Ng ◽  
J. Kenny ◽  
...  
Keyword(s):  
T Cells ◽  
Cyclosporin A ◽  
Cloning And Expression ◽  
Nuclear Factor ◽  
Activated T Cells

Facile one-pot syntheses of new C-28 esters of oleanolic acid and studies on their antiproliferative effect on T cells

2018 ◽  
Vol 73 (11-12) ◽  
pp. 417-421 ◽  
Author(s):  
Muhammad S. Ali ◽  
Ghafoor Ahmed ◽  
Muhammad A. Mesaik ◽  
Muhammad R. Shah ◽  
Mehreen Lateef ◽  
...  
Keyword(s):  
T Cells ◽  
Oleanolic Acid ◽  
Human Peripheral Blood ◽  
Bromine Atom ◽  
Alkylating Agents ◽  
Antiproliferative Effect ◽  
Activated T Cells ◽  
One Pot ◽  
Healthy Human ◽  
Peripheral Blood Mononuclear

Abstract Structure-activity relationship studies on oleanolic acid (1) have resulted in facile syntheses of its new C-28 esters 2–7 by way of one-pot reaction of 1 with a variety of alkylating agents. Oleanolic acid and its new esters were studied for their in vitro antiproliferative effect on healthy human peripheral blood mononuclear cell isolated phytohemagglutinin activated T cells. Results showed that compounds 1, 3, and 5 exhibited significant inhibitory activity on T-cell proliferation. Compound 5 was found to be the most potent, with an IC50 value of 4.249 μg/mL, among all tested compounds, and its activity could be attributed to the presence of bromine atom in the molecule.

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