Effector activity of OKT4+ and OKT8+ T-cell subsets in lectin-dependent cell-mediated cytotoxicity against adherent HEp-2 cells

Perl, András; González-Cabello, R; Làng, István; Gergely, P

doi:10.1016/0008-8749(84)90089-3

Cited by 26 publications

(21 citation statements)

References 19 publications

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“…The plates were incubated at 37 Њ C in a humidified atmosphere with 5% CO 2 for 72 h. The cultures were pulsed with 0.4 Ci 3 H-TdR 8 h before termination. Cells were harvested and 3 H-TdR incorporation was measured as earlier described (30). The results were expressed in counts per min as mean Ϯ standard error (SE) of six parallel cultures.…”

Section: Methodsmentioning

confidence: 99%

Comparative analysis of antibody and cell-mediated autoimmunity to transaldolase and myelin basic protein in patients with multiple sclerosis.

Colombo¹,

Bánki²,

Tatum³

et al. 1997

J. Clin. Invest.

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Section: Methodsmentioning

confidence: 99%

Comparative analysis of antibody and cell-mediated autoimmunity to transaldolase and myelin basic protein in patients with multiple sclerosis.

Colombo¹,

Bánki²,

Tatum³

et al. 1997

J. Clin. Invest.

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“…PBL were separated after the removal of monocytes by adherence to autologous serum-coated petri dishes (18). PBL were resuspended at 10 6 cells/ml in RPMI 1640 medium, supplemented with 10% FCS, 2 mM Lglutamine, 100 IU/ml penicillin, and 100 g/ml gentamicin in 12-well plates at 37°C in a humidified atmosphere with 5% CO 2 .…”

Section: Cell Culture and T Cell Activationmentioning

confidence: 99%

T Cell Activation-Induced Mitochondrial Hyperpolarization Is Mediated by Ca2+- and Redox-Dependent Production of Nitric Oxide

Nagy¹,

Koncz²,

Perl

2003

The Journal of Immunology

156

163

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Activation, proliferation, or programmed cell death of T lymphocytes is regulated by the mitochondrial transmembrane potential (Δψm) through controlling ATP synthesis, production of reactive oxygen intermediates (ROI), and release of cell death-inducing factors. Elevation of Δψm or mitochondrial hyperpolarization is an early and reversible event associated with both T cell activation and apoptosis. In the present study, T cell activation signals leading to mitochondrial hyperpolarization were investigated. CD3/CD28 costimulation of human PBL elevated cytoplasmic and mitochondrial Ca2+ levels, ROI production, and NO production, and elicited mitochondrial hyperpolarization. Although T cell activation-induced Ca2+ release, ROI levels, and NO production were diminished by inositol 1,4,5-triphosphate receptor antagonist 2-aminoethoxydiphenyl borane, superoxide dismutase mimic manganese (III) tetrakis (4-benzoic acid) porphyrin chloride, spin trap 5-diisopropoxyphosphoryl-5-methyl-1-pyrroline-N-oxide, and NO chelator carboxy-2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide, mitochondrial hyperpolarization was selectively inhibited by carboxy-2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (−85.0 ± 10.0%; p = 0.008) and, to a lesser extent, by 2-aminoethoxydiphenyl borane. Moreover, NO precursor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate diethylenetriamine elicited NO and ROI production, Ca2+ release, transient ATP depletion, and robust mitochondrial hyperpolarization (3.5 ± 0.8-fold; p = 0.002). Western blot analysis revealed expression of Ca-dependent endothelial NO synthase and neuronal NO synthase isoforms and absence of Ca-independent inducible NO synthase in PBL. CD3/CD28 costimulation or H2O2 elicited severalfold elevations of endothelial NO synthase and neuronal NO synthase expression, as compared with β-actin. H2O2 also led to moderate mitochondrial hyperpolarization; however, Ca2+ influx by ionomycin or Ca2+ release from intracellular stores by thapsigargin alone failed to induce NO synthase expression, NO production, or Δψm elevation. The results suggest that T cell activation-induced mitochondrial hyperpolarization is mediated by ROI- and Ca2+-dependent NO production.

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“…A total of 2500 MO3.13 cells/well of flat-bottom 96-well plates was prelabeled with [ 3 H]TdR (ICN Biomedicals) with or without 10 g/ml TALpep and allowed to form nonconfluent monolayer for 24 h. Targets were washed three times, and effectors were added for 24 h. Subsequently, plates were washed six times to remove effectors and killed target cells, and cytotoxicity was determined based on [ 3 H]TdR content of remaining viable cells, as earlier described (46).…”

Section: T Cell-mediated Cytotoxicity Assaysmentioning

confidence: 99%

“…PBMC were incubated in petri dishes precoated with autologous serum for 1 h at 37°C to remove monocytes (46). Nonadherent cells were enriched for CD8 ϩ T cells by selective depletion of CD4 ϩ cells using mAb-coated Dynabeads M-450 CD4, according to the manufacturer's instructions (catalogue 111.08; Dynal Biotech).…”

Section: Establishment Of Tal-specific Tclmentioning

confidence: 99%

CD8+ T Cell-Mediated HLA-A*0201-Restricted Cytotoxicity to Transaldolase Peptide 168–176 in Patients with Multiple Sclerosis

Niland

Bánki²,

Biddison

et al. 2005

The Journal of Immunology

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Transaldolase (TAL) is expressed at selectively high levels in oligodendrocytes and targeted by autoreactive T cells of patients with multiple sclerosis (MS). Among 14 TAL peptides with predicted HLA-A2 binding, TAL 168–176 (LLFSFAQAV, TALpep) exhibited high affinity for HLA-A2. Prevalence of HLA-A2-restricted CD8+ T cells specific for TALpep was increased in PBMC of HLA-A2+ MS patients, as compared with HLA-A2− MS patients, HLA-A2+ other neurological disease patients, and HLA-A2+ healthy donors. HLA-A*0201/TALpep tetramers detected increased frequency of TAL-specific CD8+ T cells, and precursor frequency of TAL-specific IFN-γ-producing T cells was increased in each of seven HLA-A2+ MS patients tested. Stimulation by TALpep or rTAL of PBMC from HLA-A2+ MS patients elicited killing of TALpep-pulsed HLA-A2-transfected HmyA2.1 lymphoma cells, but not HLA-A3-transfected control HmyA3.1 targets. Without peptide pulsing of targets, HLA-A2-transfected, but not control MO3.13 oligodendroglial cells, expressing high levels of endogenous TAL, were also killed by CD8+ CTL of MS patients, indicating recognition of endogenously processed TAL. TCR Vβ repertoire analysis revealed use of the TCR Vβ14 gene by T cell lines (TCL) of MS patients generated via stimulation by TAL- or TALpep-pulsed APCs. All TAL-specific TCL-binding HLA-A*0201/TALpep tetramers expressed TCR Vβ14 on the cell surface. Moreover, Ab to TCR Vβ14 abrogated cytotoxicity by HLA-A2-restricted TAL-specific TCL. Therefore, TAL-specific CTL may serve as a novel target for therapeutic intervention in patients with MS.

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Effector activity of OKT4+ and OKT8+ T-cell subsets in lectin-dependent cell-mediated cytotoxicity against adherent HEp-2 cells

Cited by 26 publications

References 19 publications

Comparative analysis of antibody and cell-mediated autoimmunity to transaldolase and myelin basic protein in patients with multiple sclerosis.

Comparative analysis of antibody and cell-mediated autoimmunity to transaldolase and myelin basic protein in patients with multiple sclerosis.

T Cell Activation-Induced Mitochondrial Hyperpolarization Is Mediated by Ca2+- and Redox-Dependent Production of Nitric Oxide

CD8+ T Cell-Mediated HLA-A*0201-Restricted Cytotoxicity to Transaldolase Peptide 168–176 in Patients with Multiple Sclerosis

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