Histamine-releasing factor/translationally controlled tumor protein (HRF/TCTP)-induced histamine release is enhanced with SHIP-1 knockdown in cultured human mast cell and basophil models

J Leukoc Biol. 2008 Oct;84(4):1151-8. doi: 10.1189/jlb.0308172. Epub 2008 Jul 14.

Abstract

Previously, we demonstrated a negative correlation between histamine release to histamine-releasing factor/translationally controlled tumor protein (HRF/TCTP) and protein levels of SHIP-1 in human basophils. The present study was conducted to investigate whether suppressing SHIP-1 using small interfering (si)RNA technology would alter the releasability of culture-derived mast cells and basophils, as determined by HRF/TCTP histamine release. Frozen CD34+ cells were obtained from the Fred Hutchinson Cancer Research Center (Seattle, WA, USA). Cells were grown in StemPro-34 medium containing cytokines: mast cells with IL-6 and stem cell factor (100 ng/ml each) for 6-8 weeks and basophils with IL-3 (6.7 ng/ml) for 2-3 weeks. siRNA transfections were performed during Week 6 for mast cells and Week 2 for basophils with siRNA for SHIP-1 or a negative control siRNA. Changes in SHIP-1 expression were determined by Western blot. The functional knockdown was measured by HRF/TCTP-induced histamine release. siRNA knockdown of SHIP-1 in mast cells ranged from 31% to 82%, mean 65 +/- 12%, compared with control (n=4). Histamine release to HRF/TCTP was increased only slightly in two experiments. SHIP-1 knockdown in basophils ranged from 34% to 69%, mean 51.8 +/- 7% (n=4). Histamine release to HRF/TCTP in these basophils was dependent on the amount of SHIP knockdown. Mast cells and basophils derived from CD34+ precursor cells represent suitable models for transfection studies. Reducing SHIP-1 protein in cultured mast cells and in cultured basophils increases releasability of the cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Basophils / cytology
  • Basophils / drug effects
  • Basophils / physiology*
  • Biomarkers, Tumor / pharmacology*
  • Cells, Cultured
  • Flow Cytometry
  • Histamine Release / drug effects*
  • Humans
  • Immunoglobulin E / pharmacology
  • Inositol Polyphosphate 5-Phosphatases
  • Mast Cells / cytology
  • Mast Cells / drug effects
  • Mast Cells / physiology*
  • Pertussis Toxin / pharmacology*
  • Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases
  • Phosphoric Monoester Hydrolases / deficiency*
  • Phosphoric Monoester Hydrolases / genetics
  • RNA, Small Interfering / genetics
  • Tumor Protein, Translationally-Controlled 1

Substances

  • Biomarkers, Tumor
  • RNA, Small Interfering
  • TPT1 protein, human
  • Tumor Protein, Translationally-Controlled 1
  • Immunoglobulin E
  • Pertussis Toxin
  • Phosphoric Monoester Hydrolases
  • Inositol Polyphosphate 5-Phosphatases
  • INPP5D protein, human
  • Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases