Histamine enhances granulocyte-macrophage colony-stimulating factor and interleukin-6 production by human peripheral blood mononuclear cells. Shlomith.
Histamine
enhances
granulocyte-macrophage
colony-stimulating human peripheral Shlomith Mor,* Ina Fabian* *Departmenz
ofCell ofBone
Abstract:
The
of cytokines was studied.
Nagler,t
Biology
and
Marrow
effect
units were
cells
ence of conditioned herent mononuclear histamine (CM-histamine)
medium cells
on
cells
but
granular
lymphocytes.
IL-6
antibodies
markedly
whereas
IL-6
activity
ceptor
in
not
and 58:
Key
Words:
T. Handzel,
ofMedicine,
Carmi
Tel Aviv
Immunobiology,
GeIlerBernstein,#
University,
Hadassah
Hebrew University Hadassah and Department ofAllergy,
histamine
effect
H1
cytokines
cells
in
activity in
GM-CSF
complete
cytokines H2
and
or
IL-6
or CM-PBS lymphocytes. H1 and H2 reblocking
J.
Hj and
of
activity. human
such
receptors.
medium
conditioned
in
CM-hista-
colony-stimulating is able to activate
and 1995.
or anti-
GM-CSF
No
to generate
IL-6 via 445-450;
radioim-
and
as GMLeukoc.
I2
production
enhances
Ramat
and
Aviv;
University
Hospital,
Medical ZamenhofSick
School,
toma
cell
cells [10]
by mouse
interleukin-6 line
treated with Histamine
to enhance the IL-6 (8.2-fold)
cytokine enhanced
resulted
cells
and
by nonadherent
blocking
(IL-i) cells of
observed the pres-
Anti-GM-CSF
CM-histamine.
antagonists
CSF Biol.
Diagnosis, Rehovot;
from nonadwith 1O M with phosphate-
found and
indicating
histamine-enhanced conclude that
mononuclear
Zeev
School Cancer
production
was in
ELISA
blocked
the
moderate,
the
levels could be detected in CM-histamine prepared from CD3, CD4, or CD8 Preincubation of CM-histamine with the We
Tumor Hospital.
(CFU-C) cultured
Using
large
was
and
prepared cultured compared
(CM-PBS).
by mononuclear
CM-PBS,
Sackler
of mononuclear in the number
kits, histamine was of GM-CSF (9.6-fold)
mine
Histology,
Transplantation
of histamine
myeloid colony-forming when bone marrow
munoassay production
Barak,
by
Jerusalem; Fund Clinic,
Israel
by subpopulations A 3.5-fold increase
saline
Vivian
lmmunology Laboratoryfor Research Institute, Kaplan
Tel Aviv,
buffered
Arnon
tDepartment jerusalem; Pediatric
factor and interleukin-6 production blood mononuclear cells
peritoneal
(IL-6)
SK-MG4,
macrophages
production
particularly
IL-i [8]. promotes
if the
proliferation
(IL-3) [11]. induces in vitro
and function; expression
it. reduces CD4+ [12] and inactivates
[13]. In the
present
mine on the mononuclear
(nonadherent CD3, histamine
cells.
study
we investigated of cytokines by mononuclear
finding
the
cytokine esses.
are
pre-
myeloid
in T cell
and augments natural killer
production cells and
the
phenotype
CD8+ cell effect
subset activity of hista-
by peripheral blood cell subpopulations
that
antagonists activity
histamine
of granulocyte-macrophage CSF) and IL-6 by peripheral phasizes
cells
unit spleen (CFU-S) CFU-S in response
alteration
and H2 receptor colony-stimulating
on
Our
glioblas-
cells, monocytes, large granular lymphocytes, and CD8 lymphocytes) and the effect H1 receptor antagonists (terfenadine
CD4,
pyrilamine) nitidine)
the
of granulocytic
(CFU-C) [9] and colony-forming and modulates proliferation ofearly
to interleukin-3 Histamine
[7] and
by
possible
secretion
(cimetidine production
increases
the
colony-stimulating blood mononuclear role
during
of
histamine and
or raby the production
factor (GMcells emin
inflammatory
of or
regulating
allergic
proc-
receptor
antagonists
INTRODUCTION Abbreviations: ing
Histamine
is known
to exert
effects
and
[1-3].
It acts on human T cells and interferon-y production
(IL-2)
to modulate
the
a variety
expression and synthesis lipopolysaccharide-stimulated clear
cells
[5] and
monocytes
production
of
of immunoregulatory of several
to inhibit interleukin-2 [4] and suppresses gene
tumor necrosis peripheral blood [6].
cytokines
It regulates
factor by mononu-
interleukin-1
units
CFU-C,
in spleen;
phosphate-buffered linked FBS,
conditioned
saline,
calcium
immunosorbent fetal
lating medium; Reprint Medicine,
bovine
factor;
assay; serum;
IL-2,
RIA.
unit-cell; medium; and
FACS,
GM-CSF,
CFU-S.
colony-form-
DPBS-CMF,
magnesium
free;
Dulbecco’s ELISA,
fluorescence-activated
granuloeyte-macrophage
interleukin-2;
IMDM,
iscove’s
enzyme-
cell
sorting;
colony-stimumodified
Dulbecco’s
radioimmunoassay.
requests:
ma
Department
Tel Aviv, P.O. Box 39040,
Journal
colony-forming
CM,
of Leukocyte
Fabian,
of Cell
Tel
Aviv
University,
Biology
and
Histology,
Sackler Ramat
School Aviv,
of
69978
israel.
Biology
Volume
58,
October
1995
445
MATERIALS
AND
METHODS
200
Growth factors and antibodies
Cl)
-j
I.
CM-PBS
L
CM-histaminj
-a
Lu Recombinant
human
Chinese
hamster
ovary
in Esclmerichia
to IL-b
CM-CSF
colt.
were
and
(CHO)
antibody
gifts
recombinant
cells, to
recombinant
GM-CSF,
of Genetics
human htiman
antibody
Institute,
lL-3
to
produced
IL-b
Cambridge,
produced
and
lL-3.
C.)
in
0
antibody
100
MA.
C.) LA. C.)
Reagents
0
z lscove’s
tnodified
Dulbecco’s
phate-buffered
salimle
purchased
purchased dihydrochloride,
Co.,
fromn
St. Louis,
Motridal,
MO.
Hertfordshire.
Grand
Island,
Rartitidine
Emlgland.
Saml Jose,
UT. were
was
CD4
and
CD8
from
serum
(FBS)
fraction from
Il-Il!, Sigma
histamine Chemical
purchased
obtained
from
MicroCELLectors
Applied
Fig.
Yerucham,
Becton
were
Immune
Effectofhistamine
.
supernatants
gift
Clara
of
were
cells
were
obtained mononuclear
7.5%
FBS
tached containing
10%
FBS.
determined
fugated
by
To cells
were
amid the
large
Percoll
fluorochrome-conjugated
440.
Berton
flow
dishes adherent
and
was
cells;
esterase
The
in
column 40%)
tile
high-density
large
granular
FACS-separated
and cells
CD4,
lymphocytes.
*1)
and CA).
The
1.
of B cells
Mononuclear
Cells
Cultured
with
Various
on CFU-C
plastic
CA).
CD3+
bating
and
instructions containing
the
buoyant pellet)
were
>95%
as
FACS
(FACS
CD16
according
pended
in
ethylenediaminetetraacetic
labeled
cells/mnl by
and
After
was
to
gently
of Histamine
of CFU-C/10’
±
1.0
CM-histamine
(10
Ni)
44.3
2.8
±
4.6
(13)’
CM-histamine
(10
M)
30.9
±
2.8
(13)c
CM-histamine
(108
M)
26.5
±
3.8
(4)C
CM-histamine
(10#{176} M)
10.0
±
1.5
(2)
CM-histamine
(1012
12.0
±
2.5
(2)
2.1
(13)
M)
CM-PBS
12.4±
rGM-CSF
(300
rlL-3
(300
rlL-6
(25U/ml) “Mean
1)M)
±
6Nuniber
(lupli(-ate). Cp
85%
room
surface.
rinses.
were
were and
cells 11-Ill
room
CD4
nonadhering
devices
devices
concentration at
by incu-
culture The
containing
1 h at
CD8
EAtS]
out
in T lym-
lymphocytes
were
CD8.
cell
monocytes.
66.4
of experimenma
or
the
pooled,
tissue
MicroCELLector for
CD4
carried
mm
15-30
suspension
DPBS-CMF
medium, were
for
selection
Sciences
or CD8.
Preparation of histamine-conditioned (CM-histamine)
tn.
Journal
were
CD4+, i0
subpopulations
nonadherent
cells
CD8
per
milliliter
(2
ml/dmsh).
.
dmshes media
were air-5%
indicated
that
day
activity.)
To
control
in which
antagonists
of
4
were
IMDM .
after
incubator. supernatants
cultures
of histamine
PBS
blood large
plated
.
10% -12 (10 -10
incubation (Preliminary showed was
added
monomiuclear at a concentration FBS -4
for optional
of
in 35-mm M)
was
4 days
studies
[data
tissue
added
at
and
37#{176}Cin not shown]
colony-stimulating
(CM-PBS).
H1 or H2 receptors
cells,
lymphocytes,
granular
with
Hmstammne
harvested CO2
cells,
lymphocytes) .
medium
(peripheral
mononuclear
and
SE.
mn(li(-ates
CD8
by negative
25-cm2
at a maximum
each
to dislodge
two 4-mi
humidified in paremitheses
of the
were
phocytes
cells”
4 ml
settle
with
>88% None
purified
Immune
CD4
incubate
into
complete
niediau,a
to
loaded
rocked
along
No.
CD3
CD8,
provided. Briefly, the 0.5% Cohn fraction
acid
allowed
incubation,
cells
Nonadherent
Concentrations
nonad-
purified
was
MicroCELLectors,
to the DPBS-CMF
low
Growth
t-ytokines
on
used
were
NA,
CD3,
isolated
(Applied
of subpopulations
the
lymphocytes Conditioned
Separation
cells
were
devices
immunomobilized
by
cells;
AD,
lymphocytes
of T lymaphocytes
covalently
(in
from
and CD8
containing
sorted
Prepared
lymphocytes;
are
in duplicate.
mononuclear
lymphocytes;
CD4
10. Results
.05.
