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Copenhagen,
Denmark
19 - 22 June 2005
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CCR1
mRNA expression in peripheral blood leukocytes
of patients with and without endometriosis
H
Xu [1,2], D Finas [1], C Banz [1], A Schultze-Mosgau
[1], AD Ebert [3], K Diedrich [1], D Hornung
[1]
[1] University of Schleswig-Holstein Campus
Luebeck
Dept. of Gynecology and Obstetrics
Luebeck, Germany
[2] Women’s Hospital School of Medicine
Zhejiang University
Department of Gynaecology
Hangzhou, China
[3] Department of Gynaecology and Obstetrics
Charite Campus
Benjamin Franklin
Berlin, Germany
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Introduction
Results published in the last decade have clearly
demonstrated that endometriosis is associated with
changes in both cell-mediated and humoral immune
responses. The major responses of women with endometriosis
include increased macrophage and B-cell number and
activity, decreased natural killer (NK) cells and
T-cell responsiveness, and increased cytokine levels
in the peritoneal fluid.
The immune alterations associated with endometriosis
are not only local, but also systemic. During the
evolution of an inflammatory response, peripheral
blood neutrophils are the first responder cells
elevated in significant numbers, followed by mononuclear
cells. Neutrophil leukocytes express chemokine receptors
including CCR1 and CXCR2. So, neutrophil-chemokine
interactions are important to inflammatory responses
depending on the repertoire of chemokines synthesized
and on the temporal
expression of chemokine receptors on the leukocytes
successfully.
RANTES (regulated upon activation, normal T cells
expressed and secreted) is found at elevated levels
in the peritoneal fluid of patients with endometriosis.
CCR1 is a CC-chemokine receptor with high affinity
for RANTES. In the present study, we investigated
CCR1 mRNA levels in peripheral blood leukocytes
of patients with and without endometriosis.
Materials and methods
The study group included 20 women of reproductive
age who had endometriosis diagnosed at laparoscopy.
The control group consisted of 20 age-matched women
without endometriosis. Blood samples were obtained
from peripheral blood of patients before anaesthesia
and laparoscopy.
Total RNA from whole blood samples was isolated
by PAXgene blood RNA kit. Reverse Transcription
PCR was done by SuperScript™ II Reverse Transcriptase
to get cDNA. Real-time PCR of CCR1 and HPRT was
done using Platinum®, SYBR®, Green qPCR
SuperMix UDG. Human CCR1 Primer Pair™ and
housekeeping gene HPRT Primer were used as primers
in Realtime PCR.
Results
Molecules of CCR1 mRNA of patients with endometriosis
were significantly increased compared to control
group. The ratio of CCR1/HPRT mRNA molecules in
patients with endometriosis (30.94±3.19)
was significantly elevated compared to women without
endometriosis (6.16±1.07) (p<0.0001).
The ratios in both lower degree stages endometriosis
(stage I and II, 23.61±2.37) and severe stages
endometriosis (stage III±IV, 33.38±4.02)
were significantly increased versus control (p<0.0001).
However, there was no significant difference of
our housekeeping HPRT mRNA between the two examined
groups.
Conclusions
Expression of CCR1 mRNA in peripheral blood leucocytes
of patients with endometriosis was significantly
higher compared to patients without endometriosis.
Both lower degree stages (stage I and II) and severe
stages endometriosis (stage III±IV) were
significantly increased versus control. Larger studies
are needed to show, if we can use CCR-1 mRNA measurement
for the diagnosis or exclusion of endometriosis
or for the follow-up of endometriosis patients.
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