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IN
VITRO
STUDIES
OF
PRESENT
DOMINGO
LUIs
ON
MUSCOLO,
the
Department
of
In vitro
Orthopaedic
studies
cytotoxic
Preliminary
transplantation
histocompatibility
there
are
bone
cells.
branes
attention
of bone
no
reports
have
not
has
on
cells
been
the
antigenic
been
isolated
components
on
from
present
in bone
surface
antigens
seem
to
of
bone
cell
great
number
the
that
generate
play
memof
immune
a key
as
bone
Muscobo,
agement
albotranspbantation
role
(Ottolenghi
in
1972;
Kawai
and Ray 1976),
immunological
of skeletal
sarcomas
(Invernizzi
and
1975),
and
possible
susceptibility
diseases
(McDevitt
also
in clarifying
help
disorders
The
and
to some
Bodmer
the
1974),
aetiobogy
of those
suspected
of being
related
present
study was undertaken
presence
of major
antigens
on bone
in
with
vitro
disparate
mine
if
cells.
individuals
they
carry
lymphocytes.
antiserum,
plantation
Isolated
bone
cells
lymphocytes
bone
containing
antigens,
might
the
cells
were
antibodies
in order
to
were
(from
cultured
genetically
species)
to deterof stimulating
exposed
against
major
transstudy
their
antigenic
MATERIALS
AND
METHODS
Lewis,
Brown-Norway
were
Bethesda,
and each,
the others
purchased
Shinya
Robert
from
equivalent
Domingo
Luis
and
Buffalo
adult
Microbiological
of human
‘1 . I).
Niuseolo.
Aires.
male
allogeneic
lymphocyte
hand,
for the presence
.
HL-A
I ntitute
(Palm
1971).
of Orthopaedics
with
and
University
of
Illinois
of transplantation
lymphocytes
On the other
and
exposed
to
their antigenic
profile.
antigens
in an active
bone cells
of serologically
were killed
by
defined
(SD)
absorbed
sera suggest
“sharing”
of
The relevance
of the surface
antigens
susceptibility
to various
orthopaedic
(Bronwill-Gilbings,
Rochester,
New
York)
fitted
with
ing
marrow
per
cent
cells,
and
transferred
dipotassium
to flasks
salt of EDTA
vated
(56
serum
in 20 milbilitres
with 5 per cent
degrees
Celsius
for
of TC
and
thirty
containing
1.5-1.7
cent heat macti-
5 per
minutes)
medium
199.
autobogous
The
flasks
were
shaken in water
gassed
CO2 in air, sealed,
and
bath at 37 degrees
Celsius
for eighteen
hOurs.
After decalcification,
the slices were washed,
ded
in
flasks
I,
Type
shaken
digested.
viability
containing
two
The
determined
60-70
per cent).
haemocytometer
8x10xml).
Fig.
for
resuspen-
milligram
of collagenase
150 units/mg)
in 10 millilitres
of medium,
and
hours
or more
until
they
were
completely
isolated
bone
cells
were
washed
twice
and
by the trypan
blue exclusion
test (usually
The cell concentration
was determined
by
and phase
contrast
microscopy
(normally
The isolated
3) or were
2-3
cells
bipolar
showed
multiple
delicate
processes
with long unbranched
processes
as
described.
Cellular
in vitro experiments
When
two populations
of lymphoid
cells from individuals
genetically
different
at the major histocompatibility
locus are
cultured
together
in vitro,
an interaction
between
the
cells
takes
place and lymphocyte
proliferation
occurs.
This reaction,
called mixed lymphocyte
culture
(MLC),
reflects
antiments,
differences
mixed
bating
between
lymphocyte-bone
to determine
lymphocyte
the
cells.
In the present
cell cultures
(MLBCC)
if bone
cells
proliferation
are
when
experiwere
capable
cultured
of stimutogether
in
vitro.
With
established
Iraumatologv,
that purpose
(Fig.
1):
Italian
hospital.
in mind
the
1) lymphocytes
U niver..itv
of
following
cultures
from
Lewis
rats+
Iienos
Ares.
Potoi
were
lym42 I 5.
M.D..
Ray.
ME).
[)epartment
of Orthopaedic
Surgery.
, Phi)..
Departhient
of Orthopaedic
Yarnaguchi
Surgery.
University.
University
School
of
of Medicine.
Illinois,
54()
South
lJhe
\Vood
(.atv.
75 Japan.
Street.
(‘hieago.
IlIinoi
#{241}l2.
U.S.A.
Requests
342
for
a
diamond
impregnated
blade.
The slices were dipped into distilled sterile water for two seconds
in order to byse any remain-
established
Associates,
The
ArgentIna.
Kawal,
D.
Fisher
Maryland.
These
strains
of rats are highly
inbred
with the exception
of Lewis and Fisher,
differs
from
at the major
histocompatibibity
locus
of the rat
Ag-B-the
Buenos
(BN),
Medicine,
to determine
stimulating
genic
Rats
and
JAPAN
the presence
antigens,
previously
to cytotoxic
UBE,
Isolation
of bone cells
The
technique
previously
described
by Bard,
Dickens,
Edwards
and Smith (1974) was used with minor modifications.
Soft tissues,
periosteum
and cartilage
were removed
from
femora
and tibiae and the marrow
was removed
by washing
with TC medium
199 (Difco,
Detroit,
Michigan).
Bone slices of approximately
150 micrometres
were cut
under continuous
irrigation
using a thin sectioning
machine
(
profile.
rats
with
(Sigma
orthopaedic
(histocompatibility)
within
the same
antigens
capable
Also
they
to auto-immunity.
to investigate
transplantation
albogeneic
manParmiani
orthopaedic
and
of
allotransplantation,
upon
KAWAI,
to investigate
cultured
RAT
OF AMERICA
School
transplantation
may not express
different
biological
phenomena
(Bach,
Bach and Sondel
1976),
and it is likely
that attempts
to define
those
antigens on bone cells could
be relevant
to clinical
situations
such
SHINYA
undertaken
albografts,
THE
STATES
Lincoln
were
focused
and/or
structures
antigens
Cell
were
such
as bone
is discussed.
homogenates
Antigenic
potential
responses.
UNITED
ANTIGENS
IN
antigens
on the cell surface.
Additional
studies
antigens
between
bone cells and lymphocytes.
much
antigenicity
ARGENTINA,
procedure
performed.
way, providing
evidence
the isolation
in a specific
cells to clinical
fields
and skeletal
sarcomata
Although
the
AIRES,
Abraham
cells
Bone
CELLS
CHICAGO,
containing
antibodies
against
results
suggest
that bone cells
form,
at least after
cytotoxic
antibodies
of bone
diseases
bone
antigens.
sera
RAY,
Surgery,
on isolated
(histocompatibility)
BONE
BUENOS
D.
ROBERT
From
TRANSPLANTATION
reprints
should
he
addressed
to
Dr
Niuscolo.
JUL
JOURNAL
OF
BONE
ANE)
JOINT
SURGERY
IN
CELLULAR
EXPERI
VITRO
STUDIES
OF
TRANSPLANTATION
343
ANTIGENS
MENTS
Lewis
HUMORAL
BN
EXPERIMENTS
Lews
BN
freeof
-
-.;:-:3
___J
marrow
bone grafts’
<complete
bone
_____________
grafts
E
a)
U.)
414.
Absorption
u R ES
Cu LT
th
BN lymphoid
cells
1
Lymphocyte
measured
by3H-Thymidine
FIG.
Cellular
experiments
Cytotoxicity
Stimulation
uptake
following
cultures
served
lymphocytes
__;p
BN
cells
bone
cells
FIG.
1
Humoral
(see text).
phocytes
from
BN rats
(MLC),
and 2) lymphocytes
from
Lewis
rats + bone
cells from
BN rats (MLBCC).
Since there
are major
histocompatibibity
differences
between
Lewis
and
BN rats,
mixed
lymphocyte
cultures
will give rise to high
stimulation.
The results
were compared
with those
obtained
in mixed
lymphocyte-bone
cell cultures.
The
against
as controls:
lymphocytes
or bone
cells
from
the same
histocompatibility
differences
and, therefore,
stimulation);
and 2) lymphocytes
from Lewis
cytes
from
BN rats + bone
cells from
BN rats
possible
inhibitory
effects
of bone
cells over
lymphocyte
cultures.
+
1) lymphocytes
rat strain
(no
no anticipated
rats+
lymphoto determine
regular
mixed
Mixed
lymphocyte
culture (MLC) and mixed
lymphocyte-bone
cell culture techniques
(MLBCC)-The
culture procedure
has
been fully described
elsewhere
(Muscobo
et a!. 1976).
Blood
was collected
by cardiac
puncture
and lymphocyte
suspenwere
prepared
by dextran
sedimentation.
After
washing,
the lymphocytes
were
resuspended
in culture
medium
(supplemented
MEM
Hank’s
base
with
10 per
cent
heat
inactivated
rat serum)
and a cell suspension
of 2.5 x 106 x ml
(85 per cent living lymphocytes)
was prepared.
The cells were
were
split into pieces
in both
2
experiments
and grafted
(see text).
into the subcutaneous
tissues
flanks.
Humoral
cytotoxicity
studies-At
intervals
tion, serum was obtained
from the immunised
studied
for
the
presence
of cytotoxic
after immunisaLewis rats and
antibodies
in the
trypan
blue exclusion
cytotoxicity
test (Tissot and Cohen 1972), using
lymphocytes
and isolated
bone cells from BN rats as target
cells.
The principle
of the cytotoxic
test is as follows:
target
cells, serum
and complement
are incubated
in vitro;
if the
serum
contains
antibodies
against
major
histocompatibibity
the target
cell surface,
complement
is fixed
and
the cell dies. The percentage
of killed cells is then determined
antigens
on
by trypan
viable
blue
cells
uptake.
Dead
are not (Figs.
Results
were
expressed
are
-
stained
by trypan
blue,
5).
as cytotoxicity
- control
sample
100
cells
3 to
index:
xlOO
control
sions
The
five
Lewis
(with
no
mean
rats
major
cytotoxic
grafted
percentage
with
obtained
bone
histocompatibility
from
Lewis
differences;
with
sera
from
or Fisher
rats
not
included
in
Figure
2) tested
against
lymphocytes
or bone cells from BN
rats (4 per cent and 41 per cent cytotoxicity
respectively)
mixed
and cultured
in 0.2 millilitre
microplates.
After
120
served
as control
values.
Significance
levels were calculated
hours
of incubation
at 37 degrees
Celsius
in humidified
5 per
t test.
cent CO2 air atmosphere,
1 pCi of 3H-Thymidine
(Sp. ac 6#{149}7by the Student’s
Ci/Mmobe)
was added to each culture.
Eighteen
hours
later,
Absorption
studies
(Fig. 2)-Absorption
is the removal
of
the cultures
were precipitated
onto glass fibre paper and the
antibody
from an immune
serum by treatment
with particulate
lymphocyte
proliferation
assayed
by determining
the 3Hantigen
homologous
to that antibody,
followed
by centriThymidine
uptake
using
liquid
scintillation
spectrometry.
fugation
and separation
of the antigen-antibody
complex.
Varying
concentrations
of bone
cells
were
used
in the
Serum
obtained
from Lewis rats grafted
with bone from
MLBCC,
ranging
from 0.4 to 1 .2 x 106 x ml.
BN rats was divided
in two groups:
one was untreated,
a
The results were expressed
as the mean count per minute
second
was
absorbed
with
BN
lymphoid
cells.
The
cytotoxic
(
± standard
Humoral
error)
experiments
Immunisation
with complete
marrow
(cortex
VOL.
9
59-B,
of four
No.
replicate
(Fig.
cultures.
2)
animals-Twenty
Lewis rats were grafted
bone
(cortex
and marrow)
or bone free of
minus
marrow)
from BN rats.
The bones
of
3. AUGUST
1977
activity
of both serum groups was then tested against
cytes and bone cells from BN rats as described.
The
purpose
of this part of the experiment
determine
if lymphocytes
and
bone
cells
share
gens.
If so, both should
show none or at least
cytotoxicity
when tested with the absorbed
sera.
bymphowas
surface
to
anti-
diminished
344
D.
L.
MUSCOLO,
S.
.
KAWAI
AND
R.
D.
RAY
.
.77
.
,)i
3
FIG.
Figure
3-Isolated
medium,
uptake
and
Absorption
was
cell
(Wright,
Hargreaves,
1973) by incubation
with
2 x 1O
Celsius.
After
stored
medium.
performed
serum
and
culture
contrast.
excluding
undiluted
Specificity
samples
with
in
blue.
Phase
lymphocytes
and HeblstrOm
at 37 degrees
suspended
of trypan
two living
Bernstein
removed
FIG.
bone
shows
lymphocyte
;...
at
-20
controls
lymphoid
lymphoid
(x
the
cells
for
Bansal,
sixty
Celsius
until
5
FIG.
(x
1200.)
Figure
4-Isolated
hone
cell
suspended
in
showing
uptake
of trypan
blue
in culture
medium.
Phase
contrast.
culture
by one
( x 900.)
dead
Li
of
minutes
the serum
was
used.
were done by absorbing
cells from “third party”
4
conrast.
1200.)
Figure
5-Photomicrograph
dye.
The cells are suspended
of2 milbilitres
centrifugation,
degrees
Phase
(I)
+1
aC
8000
Li
some
Buffalo
sera
rats.
Fa-
Li
z
RESULTS
Cellular
in vitro
Mixed
major
rats+
high
experiments
(Fig.
6)
lymphocyte
cultures
between
rats that differ at the
histocompatibility
locus (lymphocytes
from Lewis
lymphocytes
from
BN rats)
gave,
as expected,
and
(7898
reproducible
C.P.M.
lymphocyte-bone
combination
from
BN
levels
±SE
cell
(lymphocytes
rats)
did
of lymphocyte
On
556).
the
cultures
from
not
show
000
I
stimulation
other
with
Lewis
2000
I.;-
hand,
mixed
the same
strain
rats + bone
cells
lymphocyte
stimulation
in
any of six consecutive
experiments
(49 C.P.M.±SE
7).
These
values
were practically
the same as those obtained
in the
control
experiments
lymphocytes
or bone cells
cultured
therefore,
(with
with
from
which
the same
no histocompatibility
no expected
stimulation)
This
suggests
isolation
procedure
culture
conditions
lymphocytes
The
in
isolated
bone
some
ruled
inhibitory
effect
over lymphocyte
out.
Mixed
lymphocyte
cultures
cells
were
202)
with
from
added
(lymphocytes
and bone
lymphocyte
and no significant
regular
mixed
Lewis
rats
Serum
raised
BN
differences
lymphocyte
+ lymphocytes
in Lewis
may
have
stimulation
in which
was
bone
Lewis
rats +
from
cells from
stimulation
Humoral
cytotoxicity
(Fig. 7 and Table
I)
cells
rats)
(7315
showed
C.P.M.
high
±SE
were found
(P >050)
cultures
(lymphocytes
from
BN
rats)
rats
with
bone
grafts
culture
(MLC)
and mixed
lymphocyte-bone
cell
results.
Cultures
were harvested
after
120 hours
after
hours
pulse
with
3H-Thymidine.
Values
for
3H-Thymidine
uptake represent
the lymphocyte
stimulation
in each
culture.
Note
that bone
cells alone
were
incapable
of stimulating
albogeneic
lymphocytes
in vitro.
Lymph. = lymphocytes;
Lewis=
(MLBCC)
eighteen
Lewis
from
grafted
BN
rats
animals)
rats;
BN = Brown
(fifteen
showed
Norway
rats.
antisera
from
nine
different
69 ± SE 2 cytotoxic
index
over
BN
lymphocytes
and
(P<
0.#{216}J1)
Serum
raised
37 ± SE
in Lewis
4 over
rats
with
BN
bone
bone
grafts
cells
free
of
marrow
from BN rats (twelve
antisera
from six different
grafted
animals)
showed
60± SE 5 cytotoxic
index over
BN lymphocytes
and
31 ± SE 3 over
BN bone
cells
(P <
tested
from
complete
lymphocyte
0001).
The
experiments
6
FIG.
Mixed
culture
that
bone
cells,
at least
after
the
that
was
performed
and
under
used,
did not stimulate
allogeneic
in vitro.
possibility
that
lymphocytes
levels
of
and,
49
CULTURES
lymphocytes+
rat strain
were
differences
(Fig. 6).
,,(
___________
specificity
over
Buffalo
“third
rats,
of these
two
groups
party”
lymphocytes
and no significant
of antisera
was
and bone
cytotoxicity
cells
was
found.
THE
JOURNAL
OF
BONE
AND
JOINT
SURGERY
IN
VITRO
STUDIES
OF
TRANSPLANTATION
These
plantation
70
60
Li
(I,
r?
rLL&I
Sero
...
Sero
raised
with bone
free
of marrow
raised
complete
. . .
+1
with
bone
grafts.
toxic
levels
Absorption
x
Li
0
Absorption
lymphoid
z
40
carried
>-
I-
0
><
0
BN
20
cytotoxic
control
bone,
found
tests
(Table
anlayses
and bone
were
Bone
was
Cells
cally
(unabsorbed:
values
as
cytotoxicity
-
sample
-
100
It
been
responsible
a limited
x 100.
control
CvioToxlcrrv
OF HUMORAL
BONE
Significance
levels
CELLS
AND
were
grafts
serum
from
after
receiving
BN rats
INDICES
OBTAINED
LYMPHOCYFES
calculated
using
bone
bone
on
the
effect
from
over
Control
sera
from
Lewis
five different
BN lymphocytes
Finally,
Lewis
marrow
the
obtained
Fisher
rats)
showed
and 0±SE
cytotoxic
rats with
BN complete
bone were compared
Lewis
rats
rats
(twelve
59-8,
No.
3. AUGUST
1977
grafted
sera
at least
able
the
6
and
to
to
absorb
second.
transplantation
antigens
lymphocyte
stimulation
(Lane
and Ling 1973),
have
and
AGAINST
target
ISOLATED
t test
cells
Bone
cells
*37±4
t
1
]
T
I
t=1#{149}20
05>P>0#{149}2
31±3
indices
of
serum
with
samples
0± SE 2 cytotoxic
3 over BN bone
index
cells.
raised
bone
and BN free-of(Table
I). When
lympho-
cytes
were
used as target
cells, the differences
of the first were significant
(005
> P > 002),
cells were the target,
there
was no significant
(0.5 > P > 02)
between
both groups.
VOL.
13, 8 and
grafts
from
or
over
bone
dramati-
lymphoid
surface,
were
when
BN
dropped
that
their
first
the in vitro
distribution
Also,
over
absorbed:
indicate
60±5
bone
29;
Humoral
dropped
to
69, 67 and
taken
sera
with
t:01
t=21
005>P>002
Free-of-marrow
49 and
,
t
-
cells.
DISCUSSION
proposed
that
Lymphocytes
grafts
lymphoid
absorbed
cytotoxic
the Student
69±2w
Complete
two raised
free-of-marrow
were
antigens
for
tissue
BN
Lewis
same
I
TABLE
COMPARISON
the
the identity
of
antigens
were
6 respectively).
results
since
with
has
the
31
share
antibodies
index:
control
using
lymphocytes
(unabsorbed:
indices
for
extent,
results.
Sera raised
with complete
bone
grafts
bone
grafts
gave positive
cytotoxicity
against
cells or lymphocytes
from the donor.
Cytotoxicity
recorded
cells
transcyto-
samples,
with
BN
BN
2 and
These
cells
some
7
with
over
BN
absorption
3,
the
bone
CELLS
FIG.
Humoral
cytotoxicity
or with free-of-marrow
either
isolated
bone
bone
antisera
and one
cytotoxic
respectively).
TARGET
over
cells share
but lower
II)
absorbed
cells,
----
BN
that bone
lymphocytes,
to determine
further
cell transplantation
indices
levels
after
humoral
activity
were
65 ; absorbed:
0
Lymphocytes
indicate
with
out.
Three
different
complete
bone
30
0
I0
F-
results
antigens
antisera.
grafts.
sera.
Control
345
ANTIGENS
in favour
but if bone
difference
studies
induce
on the ability
such stimulation
(Main
1973),
1967)
in
and
incapable
brain
thyroid
cells
of cells
have
cells
(Lane,
of stimulating
other
been
than lymphocytes
reported.
Fibroblasts
(Pulvertaft
Jackson
allogeneic
have
1972),
sperm
chondrocytes
to be good
also
been
tested
cells
(Levis
(Gertzbein
stimulators.
Ling
lymphocytes
On the other
hand,
epidermal
cells
Jones
and Kountz
1971),
endotheial
Burger
recently
proved
and
and
(Main,
cells
to
Pulvertaft
1975)
are
in vitro.
Cochrum,
(Vetto
and
and
Whalen
1976) and
and
Lance
1976)
Neoplastic
bone
cells
(V#{225}nky, Stjernsw#{228}rd and Nilsonne
346
D.
L.
MUSCOLO,
S.
KAWAI
TABLE
CvroToxIc
ABsoRvrloNs
REACTIONS
PERFORMED
WITH
AND
D.
RAY
II
UNABSORBED
WITH
R.
AND
LYMPHOID
ABSORBED
CELLS
SERA
BN
FROM
RATS’
RN target
Lymphocytes
Serum
Lewis
rat
wth
grafted
Unabsorbed
Absorbed
Lewis
rat
wth
Unabsorbed
grafted
Absorbed
Lewis
rat grafted
with
BN
Cytotoxic
Unabsorbed
1972),
results
are
but
antigens
the
bone
presence
it difficult
as cytotoxicity
of the activity
of tumour-
to extrapolate
the
This
observation
difficulty
bone
should
be taken
possible
bone cell
variables
concentration
of obtaining
cells
of the
per
isolated
determined
and
this
more
bone
proved
for
antigen
it may
density
no sufficient
study,
are
(Edidin
thought
reported
tissues
that
isolated
cent
it was
to
cells
and,
on
Loghem
1966)
kidney
Belzer
and
Lance
1971),
and
their
cell
(Engelfriet,
have
lympho-
Kountz
sperm
a wide
chondrocytes
1976) were
antibodies,
antigens.
Although
presence
of
cells
(Elves
tissue
antiin this
test
distribution
studies
have
been
from different
solid
of SD transplantation
surfaces.
Heersche,
It was
Eijsvoogel
cells
(Perkins,
1975),
transplantation
cytotoxic
several
endothelial
cells
(Vetto
Gantan,
epidermal
found
and
and
Siegel,
cells
(Fellous
and
1974a;
Malseed
killed
by, or were
able
showing
cell membrane
Langer,
demonstration
serological
antigens
cells
The
indices
Czitrom,
has
with
Pritzker
and
Gross
1975),
of SD transplantation
been
no
antigens
on
reported.
that
van
Burger
Howell,
(Cooper
and
Dausset
1970),
and
to absorb,
expression
Heyner
cytotoxic
of those
evidence
in bone
for the
(Elves
isolated
bone
differences
cell lines
both
were
bone
of the humorab
cytotoxicity
for the presence
of SD
Sera
raised
in the cell-antigen
were
found,
since
significantly
cells.
cells.
experiments
transplantation
higher
Absorption
expression
cytotoxicity
against
studies
with
lymphocytes
confirmed
a “share”
of SD transplantation
antigens,
since
lymphoid
cells
were
able to absorb
most
of the antibodies
with cytotoxic effect
over bone cells.
The remnant
cytotoxicity
a
that
was
found
after
absorption
could
the possible
presence
of differentiation
antigens
on bone
cells,
as described
there
The
was
no
attempt
expression
these
been
so-called
explosive
to confirm
Frederiks,
HL-A
due to
Burwell,
of transplanted
Van
Oud
Van
Bone
Gowland
TI-fE
antigens
antigens”
has
the increasing
in close
relation
conditions.
play
a fundamental
with
and
role
Hooff,
Pena
OF
same
tissues
Dexter
BONE
in
the
and
AND
Van
between
species,
elicits
do (Chalmers
1963).
grafts
with weak antigenicity
1976) seem to preserve
JOURNAL
a great
foreign
tissues
Alblas,
Keuning,
allotransplantation,
different
individuals
within
the
transplantation
immunity
as other
freeze-dried
bone
1976; Friedbaender
speculation.
are
Termijtelen,
1975).
by
antigens
at the cell surface
is
by a chromosomab
region
called
HL-A
in the human.
Interest
in
acceptance
or rejection
(Van
Rood,
Blusse
Leeuwen
this
stimulating
“transplantation
in recent
years
evidence
that they
number
of pathological
HL-A
antigens
be explained
or tissue-specific
for other
cell lines,
of lymphocyte
and SD transplantation
genetically
determined
Ag-B
in the rat and
1959;
there
is
transplantation
6
1974b;
than
therefore,
allogeneic
(SD)
humoral
have
6
formal
Quantitative
between
not
alive”,
obtain
Eijsvoogeb
bone
membranes
using
intact
cells isolated
to determine
the presence
antigens
fibroblasts
1971),
to
29
complete
bone or with bone free of marrow
were abbe to
kill lymphocytes
and isolated
bone cells from the donor.
These
reactions
proved
to be immunologically
specific,
since control
and “third
party”
reactions
were negative.
and
to stimulate
Recently,
65
index
(see text).
Note that absorption
lymphocytes
and bone cells.
but
1972).
8
in the expen(due to the
cytes
defined
by the
2
on
stimulation.
“strength”
49
antigens
“metabolically
as essential
be possible
in their
in vitro.
Serologically
gens,
demonstrated
67
as preliminary
deleterious
effect
of collagenase
isolate
bone
cells,
on antigens
lymphocyte
Moreover,
13
provide
alive,
(Schellekens
1970).
3) The possible
and
EDTA,
used
to
3
results
evidence
million
cells
in
60 to 70 per
to be
they
were
reported
stimulation
responsible
one
2) Although
whether
has
been
lymphocyte
than
milbilitre).
cells
for
bone
cells.
due to the following
ment:
1) Inadequate
.
recorded
most
The present
studies
suggest
that isolated
bone
incapable
of stimulating
allogeneic
lymphocytes
vitro.
low
probable
makes
to normal
Absorbed
activity
was
cells removes
lymphoid
; Han
31
bone
free-of-marrow
1971
69
BN complete
bone
associated
Bone cells
BN complete
bone
S
cells
Even
(Burwell
transplanta-
JOINT
SURGERY
IN
tion
antigens
(Urist,
abbe
to
Mikulski
persistence
generate
and
of those
VITRO
an
Boyd
immune
1975),
antigens
STUDIES
OF
cells
due
gens
1975).
In
addition
duction
of
lymphocytes,
to
being
“destructive”
HL-A
circumstances
so far unclear,
antibody
called
the
graft
(Carpenter,
experimental
since
they
profile
of bone
cells,
system
may
provide
problem.
Association
of
diseases
antigens
and
psoriasis
reported
pro-
HL-A
antigens,
supporting
and
certain
genetic
origin
of these
HL-A
B27 with Reiter’s
established
infections
of
to protect
Salmonella
arthritis
and
interactions
better
with
HL-A
antigens
with
Several
reported
diseases
of bone
to be associated
HL-A
B27 with spondylitis
well accepted.
This antigen
of patients
Fries
this
occulta
Gallmeier,
mas,
leukaemias,
spondylitis
abnormal
also be
and
developments
included
in this
asymmetry
of
the
of the
group.
facet
1975),
Terasaki,
theory
(Takasugi,
Terasaki,
1973;
and
and
and
Delbon
lympho-
solid
tumours
Mickey,
Menck
Chretien,
1975)
with specific
at present
BOhme,
1972),
other
Henderson,
have
and
Rogentine,
showed
significant
HL-A
antigens.
it is unclear
how
transplanta-
relate
to the resistance
or susceptiauto-immune
and infectious
diseases
transplantation
throw
light
suggest
antigens
present
induced
sarcomas
are modified
or altered
antigens
present
in normal
cells
Baldwin
1975 ; Invernizzi
and Parmiani
Better
charactensation
histocompatibility
into the malignant
been
of
after
or
rheumatoid
Stiehm
1974)
Kuwert,
Schmidt
Tarpley,
tumour-associated
1975).
Voak
of an immuno-
and juvenile
Katz and
(Bertrams,
sarcomas
in chemically
histocompatibility
(Bowen
and
(Calm
have
the
Wetter
Thompson
that
lumbo-sacral
Spina
bifida
joints
and
in man, future
developments
in this field may
on the aetiology
of these
conditions.
Finally,
recently
presented
observations
and probably
association
of
disorders
is now
in approximately
ankylosing
Hazleman
reported.
tion HL-A
antigens
bility to neoplastic,
1975).
Some
spine
may
Reis,
antigens
1975).
(Russel,
Shultes
and Kuban
in association
with particular
myeloma
associations
Although
and joint
have
with increased
and related
is present
with
particular
of transplantation
from
the normal
of a specific
HL-A
antigen,
be in the future.
The strong
cent
immune
of
been
Twomey
been
frequency
more
will
the
also
HL-A
Johnson
conditions.
The association
disease
and acute arthritis
with
Shigebla,
Yersinia
(Brewerton
(Rachelefsky,
Multiple
et a!. 1975) in
ofthe
antigenic
understanding
would
imply
distribution
in those
patients
different
population.
recently
have
specific
and
(Bulgen,
and arthritic
have been
of a type
Jeter
1972; Langer
Further
definition
shoulder
1976)
1972)
the
tend
with
Mendell
anti(Elves
from
immune
recognition
or destruction
D’Apice
and
Abbas
1976).
There
is
evidence
suggesting
that
this may
be a
(Bonfiglio
transplantation.
95 per
for
antibodies
under
the production
“blocking”,
factor
bone
and
responsible
cytotoxic
antigens
trigger,
association
Ruderman,
Frozen
or cell debris.
However,
the role of cellular
transplantation
in the fate of bone
grafts
is far from clear
in
(Amos,
to
347
ANTIGENS
reported
response
possibly
on dead
TRANSPLANTATION
of
antigens
may
transformation
cell
surface
provide
further
of the cells.
bone
insight
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