Quantitative comparison of cytokine mRNA and inflammatory responses in cutaneous late phase allergic reactions.
The aim of this study was to quantitatively compare expression of mRNA for IL-5 and IFN-gamma with the frequency of mRNA-positive cells, total and activated eosinophils, neutrophils, lymphocytes, and vessels expressing adhesion molecules. Replicate biopsies of skin LPR to pollen antigens (Ag) and control injection sites (B) at 6 and 24 h were assessed for: (1) mRNA for IL-5 and IFN-gamma by quantitative RT-PCR (QC-RT/PCR); (2) frequency of cells expressing mRNA for IL-5 and IFN-gamma by in situ hybridization (ISH); (3) inflammatory cells and adhesion molecule expression. More mRNA for IL-5 was found in Ag- than in B-injected sites at 6 and 24 h by both QC-RT/PCR and ISH. Small amounts of mRNA for IFN-gamma were detected in Ag sites by QC-RT/PCR at 6 and 24 h, but were not significantly different than at B sites. The frequency of IFN-gamma mRNA(+)cells was higher in Ag than in B sites at 6 h. There was no correlation between the amount if IL-5 detected by QC-RT/PCR and frequency of IL-5 mRNA(+)cells by ISH. These findings also did not correlate with the degree of inflammatory responses.


CONCLUSIONS
(1) greater IL-5 than IFN-gamma deposition in Ag sites suggests Th(2)predominance in LPR; (2) lack of correlation between QC-RT/PCR and ISH findings may reflect varying mRNA content of inflammatory cells.
Zweiman
B
B
Department of Dermatology, University of Pennsylvania Medical Center, Philadelphia 19104-6057, USA.
Parrott
C M
CM
Graif
Y
Y
David
M
M
Lessin
S R
SR
D000328:Adult; D003872:Dermatitis / Q000276:immunology*; D019040:E-Selectin / Q000096:biosynthesis; D004804:Eosinophils / Q000276:immunology; D005260:Female; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology* / Q000473:pathology; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics*; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics*; D007963:Leukocytes, Mononuclear; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000276:immunology; D011058:Pollen / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012867:Skin / Q000276:immunology* / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology; D013997:Time Factors; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis
D019040:E-Selectin; D015848:Interleukin-5; D012333:RNA, Messenger; D019010:Vascular Cell Adhesion Molecule-1; D007371:Interferon-gamma