Interleukin-1 Induced Activation of Indoleamine 2,3-Dioxygenase in Human Peripheral Mononuclear Cells
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Authors
Leone, Nicolo
Issue Date
1988
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
An assay was set up to quantify the activity of indoleamine
2,3-dioxgenase (IDO) in human peripheral mononuclear cells(PMC).
Interleukin-1 (IL-1), interferon-gamma (IFN-G) and D-ala peptide T
(pep-T) were used to stimulate PMC in culture medium. Following a
44 h incubation period, PMC were pulsed with radiolabeled
tryptophan in fresh culture medium. After a period of 4 h, PMC
supernatants (SN) were aspirated, microfuged, injected into a C18
column and run through reverse-phase high performance liquid
chromatography (HPLC) in an attempt to separate tryptophan and its
catabolites. Fractions were collected at 30 s intervals and analyzed
by means of scintillation spectrometry. Counts per minute (CPM)
were plotted as a function of time for each fraction. Percent 100
specific tryptophan catabolism (%SC) was calculated for each
biological response modifier (BRM) induced and control run. These
data indicated that, although 2 separate HPLC elution systems were
used, a satisfactory separation of radiolabeled tryptophan and its
catabolites was not achieved. However, compared to the control run
for each PMC donor, some tryptophan degradation may have occured
in both IL-1 and IFN-G induced PMC, which would indicate IDO
activation. Additional chemical analysis or a different HPLC elution
method is needed to confirm whether or not IL-1, IFN-G and pep-T
induce IDO activation in PMC.
Description
iii, 31 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.