Modified skin window technique for the extended characterisation of acute inflammation in humans.
OBJECTIVE
To modify the skin window technique for extended analysis of acute inflammatory responses in humans, and demonstrate its applicability for investigating disease.


METHODS
15 healthy subjects and 5 Crohn's patients.


METHODS
Skin windows, created by dermal abrasion, were overlaid for various durations with filter papers saturated in saline, 100 ng/ml muramyl dipeptide (MDP) or 10 microg/ml interleukin-8 (IL-8).


METHODS
Exuded leukocytes were analyzed by microscopy, immunoblot, DNA-bound transcription factor arrays and RT-PCR. Inflammatory mediators were quantified by ELISA.


RESULTS
Infiltrating leukocytes were predominantly neutrophils. Numerous secreted mediators were detectable. MDP and IL-8 enhanced responses. Many signalling proteins were phosphorylated with differential patterns in Crohn's patients, notably PKC alpha/beta hyperphosphorylation (11.3 +/- 3.1 vs 1.2 +/- 0.9 units, P < 0.02). Activities of 44 transcription factors were detectable, and sufficient RNA isolated for expression analysis of over 400 genes.


CONCLUSIONS
The modifications enable broad characterisation of inflammatory responses and administration of exogenous immunomodulators.
Marks
D J B
DJ
Department of Medicine, University College London, London, WC1E 6JJ, UK. d.marks@ucl.ac.uk
Radulovic
M
M
McCartney
S
S
Bloom
S
S
Segal
A W
AW
D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000494:pharmacology; D000208:Acute Disease; D000276:Adjuvants, Immunologic / Q000494:pharmacology; D016022:Case-Control Studies; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D003424:Crohn Disease / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology; D007249:Inflammation / Q000378:metabolism*; D016209:Interleukin-8 / Q000494:pharmacology; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects / Q000473:pathology; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000166:cytology* / Q000187:drug effects / Q000473:pathology; D012884:Skin Window Technique; D014157:Transcription Factors / Q000378:metabolism
D000276:Adjuvants, Immunologic; D007155:Immunologic Factors; D016209:Interleukin-8; D014157:Transcription Factors; D000119:Acetylmuramyl-Alanyl-Isoglutamine