Immunostimulatory flagellin from Burkholderia pseudomallei effects on an increase in the intracellular calcium concentration and up-regulation of TNF-α by mononuclear cells

Ya Lei Chen, Yao Shen Chen, Hsi Hsun Lin, Chun Wen Chan, Ssu Ching Chen, Ching Hsein Chen

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Using flow cytometry analysis, the flagellin of Burkholderia pseudomallei acts as a signalling inducer, and evokes an increase in the intracellular calcium ion concentration ([Ca2+]i) in human peripheral blood mononuclear cells (PBMC). The cells with increased [Ca2+]i segregate into the live monocyte gate and not into the live lymphocyte gates. The stimulated [Ca2+]i increase can be neutralized with antiflagellin antibodies. In the absence of [Ca2+], [Ca2+]i was increased rapidly in flagellin-treated cells compared to non-flagellin-treated cells only after the addition of 1 mM CaCl2. Selective calcium antagonists were used to effectively block the [Ca2+]i signal, revealing that this signal was decreased by the addition of L-type calcium channel blockers (diltiazem, nifedipine and verapamil) and La2+ but was not changed by the addition of a T-type calcium channel blocker (flunarizine). It seemed that flagellin facilitates [Ca2+]i influx via a La2+ sensitive L-type cellular membrane channel. Furthermore, flagellin also acts as a TNF-α inducer in a time- and concentration- dependent manner when adhered mononuclear cells are treated with flagellin. This ability to induce TNF-α production was affected by the presence of [Ca2+] in the culture medium. It suggested that B. pseudomallei flagellin is an immuno-stimulatory molecule, causing an increase in [Ca 2+]i and an up-regulation of TNF-α, which may play an important role in the inflammation process.

Original languageEnglish
Pages (from-to)81-86
Number of pages6
JournalMicrobiology and Immunology
Volume51
Issue number1
DOIs
StatePublished - 2007

Keywords

  • Flagellin
  • Intracellular calcium ion concentration
  • TNF-α
  • [Ca]i

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