[Detection of Th17 cells in splenocytes of BALB/c mouse]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2009 Jun;25(6):490-2.
[Article in Chinese]

Abstract

Aim: To observe the role of IL-12 in Th17 cells differentiation.

Methods: Mouse splenocytes were prepared and cultured with alphaCD3 alone or plus different concentration of recombined mice IL-12, 3 days later, IL-17 in the culture supernatant was assayed by ELISA. In addition, Mouse splenocytes were stimulated byalphaCD3 with or without IL-12 for 48 h, then cell were washed, and restimulated by PMA and Ionomycin, the expression of intracellular IL-17 and IFN-gamma by CD4(+) and CD8(+) T cells were examined at single cell level by Flow Cytometric analysis.

Results: Th-17 cells is a independent cell population, which did not express IL-5, IL-10, IFN-gamma and Foxp3. IL-12 could induce active mouse T cells produce large amount of IFN-gamma and the differentiation toward Th1 cell. On the contrary, IL-12 could inhabit the production of IL-17 and differentiation toward Th17 cells.

Conclusion: IL-12 inhabit T cell differentiation toward Th17 cells.

Publication types

  • English Abstract

MeSH terms

  • Animals
  • CD4-Positive T-Lymphocytes / cytology
  • CD4-Positive T-Lymphocytes / metabolism
  • CD8-Positive T-Lymphocytes / cytology
  • CD8-Positive T-Lymphocytes / metabolism
  • Cell Differentiation / drug effects*
  • Cells, Cultured
  • Culture Media, Conditioned / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Flow Cytometry
  • Interferon-gamma / metabolism
  • Interleukin-12 / pharmacology*
  • Interleukin-17 / metabolism*
  • Ionomycin / pharmacology
  • Mice
  • Mice, Inbred BALB C
  • Spleen / cytology
  • Spleen / drug effects
  • Spleen / metabolism
  • T-Lymphocytes, Helper-Inducer / cytology*
  • T-Lymphocytes, Helper-Inducer / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology
  • Time Factors

Substances

  • Culture Media, Conditioned
  • Interleukin-17
  • Interleukin-12
  • Ionomycin
  • Interferon-gamma
  • Tetradecanoylphorbol Acetate