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A Szentágothai János Kutatóközpont a PTE korszerű, nemzetközi tudományszervezési és menedzsment normák szerint kialakított új intézménye, amely az élettudományi, élettelen természettudományi, valamint környezettudományi oktatás...

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Flow cytometry

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BD FACS Canto II flow cytometer

 

The digital flow cytometer contains 3 lasers (violet, blue and red) and 8 fluorescent channels.

Optical system

  • Detection of 8 FL-parameters
  • Detection of forward scatter light (FSC) with photo diode
  • Direction of side scatter light (SSC) and fluorescence signal towards PMTs

Blue laser detection (488 nm)

  • 502 LP, 530/30 nm FITC, Alexa Fluor 488, CFSE
  • 556 LP, 585/42 nm PE, PI
  • 655 LP, 670 LP nm PerCP, PerCP-Cy5.5, 7AAD, PI
  • 735 LP, 780/60 nm PE-Cy7, PE-Vio780

Red laser detection (633 nm)

  • 660/20 nm APC, Cy5, Alexa Fluor 647
  • 735 LP, 780/60 nm APC-Cy7, APC-H7, APC-Vio780

Violet laser detection (405 nm)

  • 450/50 nm PacificBlue, BV421, Alexa Flour405, BV421, eF450, Horizon V450, VioBlue
  • 502 LP, 510/50 nm AmCyan, BV510, PacificOrange, Horizon V500, Q-Dot525, VioGreen

Sample Input Sizes

  • Microtubes, 12x75 mm, and 15mL

The flow cytometer can be used to analyze all kinds of cell subsets in cell suspension, their DNA content, their cytokine and chemokine production.

High Throughput Sampler (HTS) option.

Registered users have to book an appointment in Szentagothai Research Centre website. Unregistered people should contact the device manager



FlowJo V10.

FlowJo V10. flow cytometric analysing software.



Dr. Meggyes Mátyás
researcher
meggyes.matyas@pte.hu
+36 72/536-251/31906

2020

Direct-acting antiviral treatment downregulates immune checkpoint inhibitor expression in patients with chronic hepatitis C.

Chronic hepatitis C (CHC) infection is associated with increased TIM-3, PD-1 immune checkpoint receptors expression that inhibits adaptive T cells and increases NK cell cytotoxicity against T helper cells, both resulting T cell exhaustion. Elimination of the virus with direct-acting antivirals (DAAs) may modify host immune response via altering these immune checkpoint receptors' expression. We conducted a prospective study to analyze changes in TIM-3, PD-1 and their ligands galectin-9, PD-L1 expression by peripheral blood T cell subpopulations, NK cell subpopulations, and monocytes by multicolor flow cytometry in 14 CHC patients successfully treated with 12 weeks of dasabuvir, ombitasvir, and paritaprevir/ritonavir plus ribavirin. Blood samples were collected before, at the end of treatment, and 12 and 24 weeks later. Sustained virological response (SVR) was associated with increased percentage of peripheral blood CD3+ T and CD8+ cytotoxic T lymphocytes and decreased percentage of NKbright cells. After DAA treatment, decreased TIM-3 expression by CD4+ T cells, by NKbright, and by NKT cells was found. Expression of immune checkpoint molecules' ligand PD-L1 by NK cells and by regulatory T cells and galectin-9 by NK cells and monocytes also decreased significantly at SVR. Our data suggest that DAA treatment not only inhibits viral replication but may alter host adaptive and innate immune responses. A decrease in immune checkpoint molecules and their ligands expression both on adaptive and on innate immune cells may contribute to the recovery of exhausted adaptive immune responses and to sustained virological response.

CONTACT
Dr Mátyás Meggyes
Core Facility Leader
 +36 72/536-251/31907
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Direct-acting antiviral treatment downregulates immune checkpoint inhibitor expression in patients with chronic hepatitis C.

Chronic hepatitis C (CHC) infection is associated with increased TIM-3, PD-1 immune checkpoint receptors expression that inhibits adaptive T cells and increases NK cell cytotoxicity against T helper cells, both resulting T cell exhaustion. Elimination of the virus with direct-acting antivirals (DAAs) may modify host immune response via altering these immune checkpoint receptors' expression.

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