Konference: 2006 48th ASH Annual Meeting - účast ČR
Kategorie:
Ostatní
Téma: Poster session: Basic Science and Clinical Practice in Blood Transfusion II
Číslo abstraktu: 959
Autoři: Dr. Ing. Karel Holada, Ph.D.; Jan Simak; Jaroslav G. Vostal
Three documented transfusion cases of vCJD underline the need of
better insight in blood prion protein biology. Cellular prion
protein (PrPc) plays key role in the pathophysiology of prion
diseases. Its expression by cells is necessary for amplification of
infectious prions and the disease process itself. Physiological
function of PrPc remains obscure. Its clarification may provide
important clues for the development of urgently needed blood test
and effective disease treatment. PrPc is expressed on CD34+
hematopoietic stem cells and its expression is regulated during
blood cell differentiation. Recently the importance of PrPc for
self-renewal of long-term repopulating hematopoietic stem cells was
suggested and other studies reported the protective function of
PrPc against oxidative stress and apoptosis in various cell
cultures. We previously demonstrated that human as well as mouse
red blood cells (RBC) express approximately 200 PrPc molecules /
cell (Holada et al., BJH 2000, 110, 472-80). To test if the PrPc
expression plays a role in the post-transfusion recovery and
survival of RBC we carried out transfusion study in mice. RBC
isolated from blood of wild type (WT) and PrP knockout (KO) FVB
mice were labeled in vitro by different levels of NHS-biotin. The
labeling was optimized to allow simultaneous detection of both
populations of RBC in mouse blood using flow cytometry. To exclude
the influence of different level of cell biotinylation on the
experiment outcome two mixtures of RBC were prepared. The first
contained KO RBC labeled with high and WT RBC with low level of
biotin and the second mixture contained cells labeled vice versa.
Each mixture was injected via tail vein in a group of WT mice (n=5)
and the survival of RBCs was followed. Samples were analyzed on day
1, 2, 3, 6, 9, 15, 21 and 29. The count of biotinylated RBC was
measured in comparison to 100 000 nonlabeled recipient RBC.
Simultaneously the expression of PrPc on RBC was monitored using
flow cytometry with MAb 6H4. KO RBC displayed significantly higher
first day post-transfusion recovery compared to WT RBC in both
groups of mice (81±3 % vs. 74± 3 %, P<0.005 and 90±4 % vs. 80±4
%, P<0.005). The slope of the RBC survival curve in all
individual mice during the initial 15 days was steeper for KO RBC
(mavg= - 3.44) than for WT RBC (mavg = -
2.37) suggesting the protective role of PrPc in circulating RBC.
The difference in the slope diminished during the 15 to 29 day
period which was accompanied by a 50% decrease of PrPc surface
expression on transfused WT RBC. To confirm our data the identical
experiment was carried out in a group of KO mice (n=5) transfused
with a mixture containing KO RBC labeled with low and WT RBC with
high level of biotin. Again the first day post-transfusion recovery
was higher for KO RBC (80±6 % vs. 75±6 %, P<0.05) and the
initial slope of the KO RBC survival curve was steeper in all mice
in the group. Our data suggest that PrPc expression plays role in
the post-transfusion recovery and survival of RBC. The observation
that WT RBC disappear from the circulation at lower rate than KO
RBC until their level of surface PrPc reaches 50% is compatible
with the protective role of PrPc expression on cells. Taken
together our study demonstrates that physiological role of PrPc
expression on RBC may lay in facilitating their longer survival in
circulation. (GACR 310/04/0419, MSMT 0021620806).
Datum přednesení příspěvku: 9. 12. 2006