Immunochemical characterization of the distinct monocyte cyclic AMP-phosphodiesterase from patients with atopic dermatitis.
BACKGROUND
Previous findings have suggested that the immunopathology of patients with atopic dermatitis (AD) results from altered cellular responses caused by cyclic nucleotide regulatory abnormalities. One such defect is the increased degradation of the second messenger, cyclic adenosine monophosphate (cAMP), by elevated cAMP-phosphodiesterase (PDE) activity in patients with AD.


METHODS
We used two monoclonal antibodies to identify the major PDE isoform in AD blood monocytes. We have also characterized the abnormal PDE activity by means of chromatofocusing and sucrose gradient centrifugation.


RESULTS
The chromatofocusing technique allowed the separation of a PDE-containing fraction (isoelectric point = 6.1) from AD monocytes but not from normal cells. This monocyte fraction accounted for most of the elevated leukocyte-PDE activity and was a cytosolic, cAMP-specific, low Michaelis constant, calcium-calmodulin-dependent enzyme, inhibited by the cAMP-PDE inhibitor, Ro 20-1724. The majority of the PDE activity in this chromatofocused fraction was immunoadsorbed by the solid-phase immobilized antibodies against calcium-calmodulin-dependent PDE.


CONCLUSIONS
The increased degradation of cAMP by a unique form of PDE may cause defective regulation of intracellular functions of AD monocytes, leading to the characteristic hyperreactive immune and inflammatory events. Characterization of PDE isoenzymes from different leukocyte subpopulations may allow further expansion of cell-directed therapy for inflammatory disease.
Department of Dermatology, Oregon Health Sciences University, Portland 97201.
S C
SC
Chan
D
D
Reifsnyder
J A
JA
Beavo
J M
JM
Hanifin
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000097:blood* / Q000276:immunology; D000818:Animals; D003600:Cytosol / Q000201:enzymology; D003876:Dermatitis, Atopic / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000097:blood*; D007700:Kinetics; D051379:Mice; D009000:Monocytes / Q000201:enzymology*
D007527:Isoenzymes; D015105:3',5'-Cyclic-AMP Phosphodiesterases