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New insights in heparin-induced thrombocytopenia by the use of fluid-phase assays to detect specifically platelet factor 4/heparin complex antibodies and antibody-secreting cells.
[heparin-induced thrombocytopenia]
The
key
feature
of
heparin-induced
thrombocytopenia
(
HIT
)
is
the
production
of
antibodies
(
Ab
)
against
the
platelet
factor
4
(
PF
4
)
/
heparin
complex
.
These
Ab
are
directed
against
neoepitopes
of
the
PF
4
tetramer
,
which
are
induced
by
the
complex
formation
with
heparin
.
To
study
this
humoral
immune
response
in
greater
detail
,
either
in
a
murine
immunization
model
or
in
human
blood
samples
,
reliable
and
specific
immune
assays
to
detect
specifically
Ab
against
the
PF
4
/
heparin
complexes
,
but
not
PF
4
alone
are
required
.
We
established
fluid-phase
enzyme-immunoassays
in
which
the
soluble
biotinylated
antigen
,
PF
4
/
heparin
,
is
firstly
captured
by
specific
Ab
,
and
secondly
directly
detected
with
enzyme-conjugated
streptavidin
.
The
use
of
this
fluid-phase
principle
allowed
a
higher
specificity
than
the
traditional
solid-phase
enzyme-immunoassays
in
terms
of
Ab
binding
to
murine
PF
4
/
heparin
compared
to
murine
PF
4
alone
.
This
fluid-phase
approach
applied
to
the
detection
of
specific
murine
PF
4
/
heparin
Ab
-secreting
cells
(
ASC
)
identified
the
spleen
as
the
main
lymphatic
organ
that
contributes
to
the
PF
4
/
heparin
Ab
response
in
mice
.
IgG
ASC
specific
for
PF
4
/
heparin
are
very
transiently
detectable
in
mice
,
which
might
explain
why
anti-
PF
4
/
heparin
IgG
Ab
typically
disappear
within
100
days
in
humans
.
Furthermore
,
this
fluid-phase
approach
was
successfully
transferred
to
detect
human
PF
4
/
heparin-
specific
Ab
.
T
he
fluid-phase
principle
for
the
specific
detection
of
anti-
PF
4
/
heparin
IgG
and
IgM
Ab
enables
new
and
improved
assays
for
HIT
research
in
men
and
mice
.
At
least
in
mice
PF
4
/
heparin
antibodies
are
produced
by
transient
B
cells
.
Diseases
Validation
Diseases presenting
"main lymphatic organ"
symptom
heparin-induced thrombocytopenia
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