CLINICAL RESEARCH POSTER PRESENTATION ABSTRACTS
Please note: All posters will be displayed in
Salons 10-12.
FRIDAY, 10:15-11:00 AM
Finnell JS, Yim D, Nandi A, Gunarni P, Sasagawa M,
Rosenblatt KP, Wenner CA.
Influence of Polysaccharide Krestin¨ extract of Trametes
versicolor on protein. expression in human primary peripheral blood
lymphocytes.
Bastyr University cindyw@bastyr.edu
PURPOSE: The purpose of this study was to optimize
Proteomics analysis conditions to quantify and qualify the differential
expression of immunologically relevant proteins in human primary peripheral
blood lymphocytes (PBL) treated in vitro with Polysaccharide
Krestin® (PSK), an extract of the Trametes versicolor mushroom.
Recent studies have used Proteomic pattern profiling to characterize
the differential expression of immunologically relevant proteins in
immune cells isolated from human blood upon treatment with known immune
stimulating agents. However, the application of this nascent technology
in medical mushroom research has not been fully explored.
METHODS: A total of 13 healthy human blood donors
were recruited to this study. PBL isolates from the blood of 10 volunteers
were cultured in vitro under the following conditions: untreated
(negative control); PSK (100 mg/mL); phytohemagglutinin (PHA, 1 mg/mL);
or PSK + PHA. Separate aliquots of cell lysates and supernatants were
collected at 0, 1 and 24 hr, flash frozen and stored at -80¡C until
analysis. For Proteomics testing optimization, sample sets from the
first four donors were analyzed at pH 3, 4, 5, 7 and 9 on each of four
Ciphergen Protein Chips: CM10, H50, IMAC and Q10, using a SELDI-TOF
mass spectrometer (MS) and a Pro-TOF MS. Spectra were analyzed using
ProteinChip Biomarker Wizardª by Ciphergen Biosystems, Inc., to statistically
determine differential protein expression between sample sets.
RESULTS: Analysis of spectra showed that significant
clustering of proteins in samples treated with PSK and PHA, as compared
to the untreated control, occurred in the 1 Ð 20 KDa range in both the
cell lysate and supernatant samples. Clustering was observed in over
70% of peaks in cell lysates at the 1 hour time point, and in over 50%
of peaks in supernatants at the 24 hour time point. Many immunologically
relevant proteins, including several cytokines and chemokines known
to regulate immune responses, fall within this molecular weight range.
These data suggest that PSK and PHA may both induce similar expression
patterns of immunological proteins. Determining the effects of PSK on
protein expression by immune cells in humans will lead to a better understanding
of the appropriateness of use of Trametes versicolor preparations
in promoting immunological health.
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