Transcript poster dc efis eci 2006

DECREASED PERIPHERAL BLOOD BDCA1+ AND
BDCA3+ DENDRITIC CELLS IN RHEUMATOID ARTHRITIS
AND OSTEOARTHRITIS
U.A.H
M VILLARROEL1, G ZALDIVAR1, MA SÁNCHEZ1, A HERNÁNDEZ1, JDD GARCÍA2, MJ LEÓN3, A SÁNCHEZ-ATRIO3, A
PÉREZ-GÓMEZ3, E CUENDE3, A LÓPEZ3, F ALBARRAN3, J MONSERRAT1, A PRIETO1, E REYES1, M ALVAREZ-MON1,3
(1) Laboratory of Immune System Diseases and Oncology. R&D CNB-CSIC associated unit. Department of Medicine, Alcalá de Henares, Spain.
(2) Department of Internal Medicine. “Principe de Asturias” University Hospital, Alcalá de Henares, Spain.
(3) Immune System Diseases and Oncology Service. “Principe de Asturias” University Hospital, Alcalá de Henares, Spain.
INTRODUCTION
RESULTS
Circulating Peripheral Blood Dendritic Cells (CPB-DCs)
comprise different subsets that are involved into
inflammatory response of patients suffering autoimmune
and degenerative chronic diseases.
Significant decreases in the percentages of CD19 CD14
+
+
EMA BDCA1 and CD19 CD14 EMA BDCA3 subsets were
found in RA patients and OA patients compared to HC (RA:
p<0.01 and p <0.04 respectively; OA: p<0.05 p<0.01
respectively) (Fig 2).
AIM
,2
1,4
The aim of this study is to compare the distribution of CPBDC subsets in patients with rheumatoid arthritis (RA),
osteoarthritis (OA) and healthy controls (HC).
p<0.05
1,2
p<0.01
,1
1,0
mDC 2
MATERIAL AND METHODS
mDC 1
,8
,6
Whole anticoagulated peripheral blood samples were
obtained from RA patients (N=30), OA patients (N=21) and
HC (N=7).
,4
0,0
,2
0,0
FL3H: EMA, CD14 PE-Cy5.5, CD19 PE-Cy5.5
(A)
(B)
Figure 1. Flow cytometric analysis of CPB-DCs.
The right contour plots showed the different CPB-DCs subsets, assessed in the
CD19-CD14-EMA- population.
A.Sample stain with cocktail control.
B.Sample stain with DBCA cocktail.
Abreviations: FITC, fluorescein isothiocyanate; PE, phycoerithrin; PE-Cy5.5,
phycoerithrin-cyanin 5.5; APC, allophyco-cyanin. EMA. Ethidium Bromide
Monoazide.
p<0.04
p<0.01
-,1
-,2
A
RA
OA
HC
B
RA
OA
HC
Figure 2. Distribution of mDC1 and mDC2 subsets in CPB-DCs.
Percentage distribution.
A. mDC1. anti BDCA1+
B. mDC2. anti BDCA3+
Numbers represent the significant values of U Mann Whitney test. Blue box
differences between OA and HC, orange box differences between RA and HC.
RA. Rheumatoid arthritis. OA osteoarthritis. HC. Healthy controls.
We found similar percentages
of both mDC1 and mDC2
subsets between RA and OA
patients. The percentage of
+
CD19 CD14 EMA BDCA2
DC
subset did not differ in all three
groups studied (Fig 3).
,8
,6
,4
pDC
FL3H: EMA, CD14 PE-Cy5.5, CD19 PE-Cy5.5
Blood Dendritic Cell Enumeration kit (Miltenyi Biotec –
MACS ®) was used for phenotypical assay. We used four
colours flow cytometer (FACSCalibur BD®) for data
acquisition. CPB-DCs were stained with anti-BDCA1
(mDC1), anti-BDCA2 (pDC) and anti-BDCA3 (mDC2)
monoclonal antibodies in the CD19 CD14 EMA population.
Phenotypical and statistical analyses were performed with
FlowJo (Tree Star®) and SPSS 11.0 (SPSS Inc.®) software
respectively (Fig 1).
,1
,2
Figure 3. Distribution of pDC subset in
CPB-DCs
Percentage distribution.
RA. Rheumatoid arthritis. OA
osteoarthritis. HC. Healthy controls
0,0
-,2
RA
OA
HC
CONCLUSION
We found a significant decrease in mDC subset in
RA and OA patients compare to HC.
This data also could be explained partially by the
chronic
inflammatory
response
in
both
pathologies.
The decreased percentage of mDC subset found in
these patients may reflect either increased
apoptosis of these cells or alternatively to
enhanced migration to sites of inflammation.