474

Dunham

Furthermore, differentiated TGF-3,

et al. monocytes macrophages

which

is mediated

number of receptors macrophages [42]. Macrophages such as tumor a monokine

are

much more susceptible than to the inhibitory effects of in part

for TGF-3

can be activated necrosis factor-a that

was

her of T-cell-mediated

immune

press

the

states TGF-3

synthesis

mononuclear effect on unknown.

such as has recently of TNF-a

cells

ticoids. However, it has any of the immunosuppressive equivalent to the widely corticoids.

Furthermore,

addition suppressive

of interferon-rny effects of

diated mals

and

isolated

RPMI

and sup-

but

[41]

on

the

used

suppressive

effects

this

report, the

recently

shown

that

(IFN-y) in vitro can both glucocorticoids

reverse [40]

the and

of toxic

we

iodination doses

metab-

can

partially cell-me-

by neutrophils of dexamethasone

on the synthesis

show of

with

of TNF-a

that

porcine

TGF-132

from

LPS-stimulated

Porcine TGF-132 antiinflammatory

supis

as glu-

dexamethasone, which is more suppressive natural glucocorticoid cortisol. Recombinant in combination with LPS, primes macrophages

even in the presence of moderately TGF432 or glucocorticoids.

suppressive

and

of

human

U/mg),

Cell The

AND

Lines murine

fibrosarcoma

was a gift from Dr. Research, Frondheim,

WEHI

164 clone

Terje Espevik (Institute Norway). These cells

tained in RPM! 1640 with tories, Grand Island, NY), prepared by supplementing

13 cell

line

of Cancer were main-

L-glutamine (Gibco Laboraand complete medium was with 2 g/liter sodium bicar-

were Limof 25 MA). RPM!

secreting leuType Culture

maintained

in complete

Products

TNF-a

(rHuTNF-a; TNF-3

monoclonal

5.0

(rHuTNF-3; anti-HuTNF-a

X

iO

1 .2

x

(>5

X

neutralizing units/mi), and rabbit polyclonal antiHuTNF-3 (2.90 x i0 neutralizing units/ml) were gifts from Dr. John Kaumeyer of Genentech, Inc. (South San Francisco, was kindly Limited of 5-10

CA). Recombinant supplied by Dr.

(Basel, Switzerland), x lO U/mg of

rPoIFN-y, as well from Dr. Bernard porcine platelets,

MN),

1 ml 4 mM

serum albumin dexamethasone,

porcine IFN-’y (rPoIFN-y) Rolf Steiger of Ciba-Geigy and protein.

had a specific activity Rabbit antiserum to

as rabbit preimmune serum, were gifts Charley [9]. TGF-32, purified from was purchased from R & D Systems,

(Minneapolis,

and HC1

prepared

by reconstituting

containing

1 mg/ml

bovine

(BSA). and LPS,

Hydrocortisone-21 phenol extracted

succinate, from Escher-

ichia co/i, were purchased (St. Louis, MO). Stock prepared; hydrocortisone-2

from Sigma solutions of 1 succinate

Chemical Co. 1 mg/mI were was diluted in

dilutions calf ml).

salt solution in 200-proof

were

prepared

(HBSS) and ethyl alcohol.

concentration ments (40

of nM)

Porcine

that were used were observed

HBSS

supplemented

ig/ml

streptomycin

No

in preparing TGF-132 in any experiment.

isolated

following with was

from

lungs

or dex-

of 5-month-old

exsanguination.

500

U/mI

poured

into

One penicillin the

of the lungs, the medium tubes that were maintained

lavage

discarded

was

5%

used in these experisignificant effects of

gentle massaging into 50 ml conical fluid

containing

Macrophages were

pigs

dexamethaSubsequent

UT; endotoxin -

1’

20

represent

superscripts

are

.05).