Analysis of interleukin 12 protein production and mRNA expression in mice exposed topically to chemical allergens

Toxicology. 1999 Jan 1;132(1):57-66. doi: 10.1016/s0300-483x(98)00137-1.

Abstract

Interleukin (IL) 12 is a heterodimeric cytokine which stimulates IFN-gamma production and promotes the development of type 1 T helper cells and T cytotoxic cells from their respective precursors. We have described previously that contact allergens such as 2,4-dinitrochlorobenzene (DNCB), and respiratory allergens such as trimellitic anhydride (TMA) induce discrete type 1 and type 2 cytokine secretion patterns, respectively, following repeated topical exposure of BALB/c strain mice. Under such conditions, we have now examined production by draining LNC of the inducible subunit of IL-12 (p40) and p40 and p35 subunit mRNA expression. Cultured LNC prepared from mice treated with DNCB secreted significantly more IL-12 p40 protein than did TMA- or vehicle-activated LNC, such differences becoming more pronounced as the duration of culture increased. Maximal levels of mRNA expression of the IL-12 p40 subunit were observed after 6-24 h of culture in all treatment groups and declined thereafter. Somewhat higher levels were induced following exposure to DNCB, these differences only reached statistical significance at 24 h of culture. Expression of this subunit by LNC from all treatment groups declined with time in culture. Levels of IL-12 p35 mRNA expression were comparable in cultured LNC prepared from allergen and vehicle treated mice and remained constant throughout the entire culture period. These data indicate that the divergent T cell cytokine responses seen in response to contact and respiratory allergens are associated with differential production of IL-12 p40 protein in the absence of substantial changes in the expression of mRNA for either subunit.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetone / pharmacology
  • Administration, Topical
  • Allergens / pharmacology*
  • Animals
  • Cells, Cultured
  • DNA Primers / chemistry
  • DNA-Binding Proteins / metabolism
  • Dinitrochlorobenzene / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Interleukin-12 / biosynthesis*
  • Interleukin-12 / genetics
  • Lymph Nodes / cytology
  • Lymph Nodes / drug effects*
  • Lymph Nodes / metabolism
  • Mice
  • Mice, Inbred BALB C
  • NADPH Oxidases
  • Olive Oil
  • Phthalic Anhydrides / pharmacology
  • Plant Oils / pharmacology
  • Proteins*
  • RNA, Messenger / biosynthesis*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Allergens
  • DNA Primers
  • DNA-Binding Proteins
  • Dinitrochlorobenzene
  • Olive Oil
  • Phthalic Anhydrides
  • Plant Oils
  • Proteins
  • RNA, Messenger
  • Acetone
  • Interleukin-12
  • trimellitic anhydride
  • NADPH Oxidases
  • NCF4 protein, human