Assessment of preferential Th1 or Th2 induction by low-molecular-weight compounds using a reverse transcription-polymerase chain reaction method: comparison of two mouse strains, C57BL/6 and BALB/c

Toxicol Appl Pharmacol. 2001 Nov 15;177(1):38-45. doi: 10.1006/taap.2001.9286.

Abstract

The aim of this study was to examine whether the RT-PCR method for various Th1/Th2 cytokines is appropriate for determination of response to allergens using C57BL/6 and Balb/c mice, which are known to preferentially demonstrate Th1 and Th2 responses, respectively. To this end, both strains of mice were sensitized by skin painting with the contact allergen dinitrochlorobenzene (DNCB) or the respiratory allergen trimellitic anhydride (TMA). We used the sensitizing protocol adopted by Kimber and coworkers (Toxicology 103, 63-73, 1995). At various time points after the last application, the levels of mRNA expression for Th1-type cytokines IFN-gamma, IL-18, and IL-12p40, as well as receptor IL-18R, and the Th2-type cytokine IL-4 and the receptor ST2L, in lymph nodes were measured. The results suggest that differential expression of IL-12p40 and IL-4 mRNA after 24 h allows clear discrimination between DNCB and TMA in C57BL/6 mice, more obviously than in Balb/c mice. Furthermore, to examine this method, C57BL/6 mice were exposed to OXA, DNFB, and TNCB (Th1-predominant allergens) or PA, TDI, and MDI (Th2-predominant allergens). Elevation of IL-12p40 expression was significant with the Th1 inducers, while the level of IL-4 was higher with Th2-predominant allergens. The results of the present study demonstrate, for the first time, that differential expression of IL-12p40 and IL-4 in C57BL/6 mice may be useful as a parameter for assessing influence of contact and respiratory allergens.

Publication types

  • Comparative Study

MeSH terms

  • Allergens / chemistry
  • Allergens / classification
  • Allergens / toxicity*
  • Animals
  • DNA Primers / chemistry
  • Dinitrochlorobenzene / toxicity
  • Dinitrofluorobenzene / toxicity
  • Interleukin-12 / biosynthesis
  • Interleukin-12 / genetics
  • Interleukin-4 / biosynthesis
  • Interleukin-4 / genetics
  • Isocyanates / toxicity
  • Local Lymph Node Assay
  • Lymph Nodes / drug effects
  • Lymph Nodes / metabolism
  • Lymph Nodes / pathology
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Molecular Weight
  • Oxazoles / toxicity
  • Phthalic Anhydrides / toxicity
  • Picryl Chloride / toxicity
  • RNA, Messenger / biosynthesis
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Species Specificity
  • Th1 Cells / drug effects*
  • Th1 Cells / metabolism
  • Th2 Cells / drug effects*
  • Th2 Cells / metabolism
  • Toluene 2,4-Diisocyanate / toxicity

Substances

  • 4-ethoxymethylene-2-phenyl-2-oxazoline-5-one
  • Allergens
  • DNA Primers
  • Dinitrochlorobenzene
  • Isocyanates
  • Oxazoles
  • Phthalic Anhydrides
  • RNA, Messenger
  • Toluene 2,4-Diisocyanate
  • Interleukin-12
  • Interleukin-4
  • trimellitic anhydride
  • 4,4'-diphenylmethane diisocyanate
  • Dinitrofluorobenzene
  • phthalic anhydride
  • Picryl Chloride