Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY, 1 4 ( 3 ) , 4 5 1 - 4 6 1 ( 1 9 9 2 )
S E R U M TUMOR N E C R O S I S FACTOR A C T I V I T Y I N INFLAMMATORY BOWEL D I S E A S E M . Maeda, N . Watanabe, H. Neda, N . Y a m a u c h i , T. Okamoto, H. Sasaki, Y. T s u j i , S . Akiyama, N . T s u j i and Y . N i i t s u Department of Internal M e d i c i n e ( S e c t i o n 4 ) , Sapporo Medical C o l l e g e , South-1, West- 16, Chuo-ku, S a p p o r o 0 6 0 , Japan
ABSTRACT Serum tumor necrosis factor (TNF) levels i n 33 p a t i e n t s with inflammatory bowel d i s e a s e (IBD) were measured by using a s e n s i t i v e enzyme immunoassay. Four o f five Crohn's diseases (CD) and n i n e o f twenty e i g h t ulcerative c o l i t i s (UC) had elevated l e v e l s of serum TNF. I n active CD o r UC, a greater fraction o f p a t i e n t s studied had s i g n i f i c a n t l y increased serum TNF levels (3/3 f o r CD and S / l 1 f o r UC). Production of TNF by peripheral b l o o d m o n o c y t e s when stimulated b y lipopolysaccharide was a l s o increased in t h e s e patients and correlated with t h e i r serum TNF levels. These rusults suggest that TNF may have s o m e p a t h o e t i o l o g i c a l meaning i n IBD.
INTRODUCTION Many immunological disturbances are associated with (1-6).
Increased h e l p e r T c e l l s (CD4 p o s i t i v e )
IBD
and decreased
suppressor T c e l l s (CDS p o s i t i v e ) populations in t h e peripheral circulation were found frequently i n UC (1).
In addition, enhanced
expression o f h u m a n leukocyte antigen (HLA) was recently correlated with
IBD
(2).
Furthermore,
451 Copyright @ 1992 by Marcel Dekker, Inc.
immune
cyt o k i n e s
including
MAEDA ET AL.
452 TABLE 1 Patients Profiles
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
uc
CD
Sex
28 M13, F15
5 M4, F1
Age(y r) Disease activity
19-42 active remission
23-33 active remission
3 2
Extent of lesion
total colitis left-sided colitis proc titis
ileocolitis colitis
3 2
No of cases
11 17 7 15
6
interleukin-1 (IL-1) (3), interferon (IFN) (4, 5 ) and TNFare assumed to be involved in t h e hyperimmune s t a t e of IBD. I n the present p a p e r , we describe serum TNF levels and TNF production by circulating monocytcs i n p a t i c n t s w i t h IBD.
PATIENTS AND METHODS Human Serum Samoles We studied 3 3 p a t i e n t s ( 1 6 male and 1 7 female, aged 1 9 t o 46 years) w i t h IBD.
The p a t i e n t s had a h i s t o l o g i c a l l y
confirmed
diagnosis of IBD. The p a t h o l o g y corresponded t o UC i n 2 8 cases (active stage cases (active
:
11 cases, remission s t a g e : 3, remission
:
2) (TABLE1).
Discase c o n t r o l s were as follows polyp ( n = 3 ) , ischemic
17 cases), and CD i n 5
c o l i t i s (n=2),
c o l o n cancer ( n = l 9 ) , colon infectious c o l i t i s
(n=8),
pneumonia ( n = 4 ) , c h o l e c y s t i t i s (n=3), and h e p a t i t i s ( n = 3 ) . Twenty healthy age-matchcd laboratory workers s e r v e d a s normal c o n t r o l s .
453
TUMOR N E C R O S I S FACTOR I N BOWEL D I S E A S E
Serum samples were inactivated by heating a t 56C for 3 0 m i n , and
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
were stored a t -70 “c un ti1 assayed. Recomb in an t Human TNF Recombinant human TNF (rhTNF), produced in Escherichia coli and purified (99.9%) was generously provided by Asahi Chemical Ind. C o . , Ltd. (Tokyo, Japan) (7). The rhTNF had a s p e c i f i c a c t i v i t y of 2.37 X l o 6 units (U)/mg protein a s determined b y i t s c y t o t o x i c activity a g a i n s t TNF s e n s i t i v e L-M (mouse tumorigenic f i b r o b l a s t cells, ATCC CLL 137, American Type Culture Collection, R o c k v i l l e ,
MD.) (8). A TNF activity unit (U) was defined as t h e amount of TNF g i v i n g 50% L c e l l survival as determined by dye-uptake (7). ELISA (Enzyme Linked Immunosorbent Assay) Serum TNF levels were measured by a double antibody enzyme linked immunosorbent assay (ELISA, Asahi Chemical Ind. C o . , Ltd.) (9). Microtiter plates (Dynatech, Immulon 11, 96 wells) were coated with rabbit anti-rhTNF IgG by o v e r n i g h t incubation with a n t i b o d y at 4°C. The plates were then washed 5 times with p h o s p h a t e buffered s a l i n e (PBS) c o n t a i n i n g 0 . 2 % Triton X-100. Next, aliquots o f serum from p a t i e n t s were added to t h e wells. Serially diluted rhTNF i n dog plasma was included to establish a standard curve. The p l a t e s were then incubated with serum samples at 4°C for 2 4 h . After washing w i t h PBS c o n t a i n i n g 0 . 2 % Triton X-100, peroxidase conjugated
anti-TNF a n t i b o d y was added. The plates were then incubated for 6 h at r o o m temperature. After washing, they were incubated with p-n itro-phen ylphosphat e substrate a t room temperature for 1 O m i n . The absorbance of product was measured a t 492nm using an ELISA reader (SLT-Lab Instruments, Austria). TNF levels were detected i n a dose dependent manner a t concentrations between 0 . 2 and 2.0 U/ml. The determinations were made in triplicate. TNF i n
serum was
The detection limit of
about 0.2U/ml corresponding t o
concentration of 10Opg/ml by t h i s assay.
a rhTNF
454
MAEDA ET A L .
Isolation a n d c u l t u r e o f Human Peripheral Blood Monocytes Mo n o n ucl ear cell s were i so I a ted from h ep ari ni zed p eri ph er a1 Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
venous b l o o d (1OU heparin/ml b l o o d ) b y Ficoll-Isopaque gradient density centrifugation. The m o n o c y t e fraction was isolated using macrophage separation p l a t e (MSP-P, Otsuka Assay Laboratories, Osaka, Japan).
The monocyte concentration was adjusted t o 5 X
106/ml i n Eagle's minimal e s s e n t i a l medium (EMEM) with
10%
h e a l - i n a c t i v a t e d f e t a l b o v i n e serum (FBS, Flow L a b . , Australia), 1 0 0 U/ml of p e n i c i l l i n , 1OOp g/ml of streptomycin and 10mM HEPES
(pH7.2). These c e l l s were t h e n incubated for 20h in t h e presence o r absence o f 1 0 p g/ml lipopolysaccharide (DIFCO, LPS B , E. c o l i , 01 27
B8) a t 37°C and 5 % CO, in humidified a i r . The supernatants
were collected after 2 0 h a s conditioned media, filtered through a 0 . 2 2 p m m i l l i p o r e filter,and kept frozen at -20°C until assayed.
Bioasaay for TNF
To measure t h e TNF a c t i v i t y i n a b i o a s s a y , standardized serum samples and conditioned media of monocytes from IBD patients were incubated with Actinomycin D (final concentration l p g/ml) treated L-M c e l l s at 37°C for 48h in 5 % CO,.
C y t o t o x i c activity was t h e n
a s s e s s e d b y the dye uptake method ( 1 0 , l l ) .
RESULTS Serum Levels o f TNF i n Patients with IBD --TheserumTNFlevels of all20 healthy subjects were below detection l i m i t . W i t h IBD, on t h e o t h e r hand, p o s i t i v e e l e v a t i o n s were observed i n 8 0 % (4/5) of p a t i e n t s with CD and 31% ( 9 / 2 8 ) of patients with UC (FIG. 1 ) . The percentages of p o s i t i v e e l e v a t i o n s in p a t i e n t s with t h e a c t i v e stage were as much as 1 0 0 % ( 3 / 3 ) for CD and 7 2 . 7 % (8/1 1) for UC. I n t h e s e i n s t a n c e s , serum levels of TNF were high at 0 . 4 2 - 0 . 8 6 U/ml for C D a n d 0 . 2 - 2 . 1 2 U/ml f o r UC.
455
TUMOR N E C R O S I S FACTOR I N BOWEL D I S E A S E
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
A 2.12
A A
_ _ _ _ *_ _ _ _ - _ _ _ _ _ - _ 0.0.
Healthy Crohn's Ulcerative Infectious Colon Other Colon Other Control Disease Colitis Colitis Cancer Disease Inflammatory (n=20) (n=5) (n=28) (n=8) (n=19) (n=5) Disease (n=10)
FIG. 1. Serum TNF concentration i n p a t i e n t s with c o l o n diseases. In I B D p a t i e n t s , A a n d A represented a c t i v e and remitted disease, respectively. Serum samples listed under o t h e r colon diseases were taken from p a t i n t s with ischemic c o l i t i s (m) and p o l y p s
(a).
As f o r o t h e r diseases o f t h e c o l o n , increased serum TNF l e v e l s
were seen i n 26.7% ( 3 / 8 ) p a t i e n t s with infectious c o l i t i s , 9 . 5 % (2/19) o f p a t i e n t s with c o l o n cancer,and 1 0 0 % (2/2) of p a t i e n t s with ischemic c o l i t i s a s shown i n other colon disease group o f F i g . 1 , however, serum TNF levels i n p a t i e n t s with these diseases were lower than t h o s e in p a t i e n t s with a c t i v e IBD.
The serum TNF in
these c o n t r o l s was n o more than 0.27 U/ml (patients with colon cancer had t h e highest levels). The three p a t i e n t s with c o l o n p o l y p s showed no p o s i t i v e result. Inflammatory diseases i n s i t e s other than t h e large i n t e s t i n e ( h e p a t i t i s , 3 cases; c h o l e c y s t i t i s , 3 c a s e s ; pneumonia, 4 c a s e s ) , a
MAEDA ET AL.
456 2.12
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
0.9-
>-
It7
(pre ope)
0.80.7-
53
.- 0.6Y
P
0.5-
u C
2 z k
0.4-
IJ.
0.3-
therapy
therapy
FIG. 2. Serum TNF concentration i n IBD p a t i e n t s before and after therapy. I n IBD p a t i e n t s , 0 a n d 0 represented UC and CD, respectively .
p o s i t i v e e l e v a t i o n s were obtained only in two cases o f pneumonia; the serum TNF levels i n t h e s e cases were l o w a t 0 . 2 6 U/ml and 0 . 3 1 U/ml compared to t h o s e in cases o f UC and CD. Relationship between P a t h o l o g i c Features o f IBD and Serum TNF Serum l e v e l s o f TNF were measured i n t h r e e CD p a t i e n t s and s i x UC p a t i e n t s before and a f t e r treatment. Of t h e p a t i e n t s with UC, o n e case received corticosteroid, a n o t h e r underwent surgery, and t h e other four cases received sulfasalazine. Of t h e p a t i e n t s with CD, o n e case received corticosteroid, a n o t h e r sulfasalazine o r elemental diet therapy. Regardless o f t h e treatment regimen, serum TNF decreased below t h e detection limit(O.2U/ml) i n b o t h CD and UC (FIG. 2), and correlated well with r e m i s s i o n . Furthermore, i n o n e C D patient between serum TNF levels,
there was g o o d correlation
C-react v e p r o t e i n ,
(Crohn's disease a c t i v e index) (FIG. 3 )
and the CDAI
- -6oi\ y-yF:200
457
TUMOR N E C R O S I S FACTOR I N BOWEL D I S E A S E ESR
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
SERUM TNF (Ufml)
70
CDAl
(Ihr)
50
ESR
40
.
10
.
0
J
150
0.6
Q
CRP
0 0.
--
Da :e
lj ar
.inn
0.54 ...
.11
-Eo4++NF
2/27
I
50
---o
0
IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY, 1 4 ( 3 ) , 4 5 1 - 4 6 1 ( 1 9 9 2 )
S E R U M TUMOR N E C R O S I S FACTOR A C T I V I T Y I N INFLAMMATORY BOWEL D I S E A S E M . Maeda, N . Watanabe, H. Neda, N . Y a m a u c h i , T. Okamoto, H. Sasaki, Y. T s u j i , S . Akiyama, N . T s u j i and Y . N i i t s u Department of Internal M e d i c i n e ( S e c t i o n 4 ) , Sapporo Medical C o l l e g e , South-1, West- 16, Chuo-ku, S a p p o r o 0 6 0 , Japan
ABSTRACT Serum tumor necrosis factor (TNF) levels i n 33 p a t i e n t s with inflammatory bowel d i s e a s e (IBD) were measured by using a s e n s i t i v e enzyme immunoassay. Four o f five Crohn's diseases (CD) and n i n e o f twenty e i g h t ulcerative c o l i t i s (UC) had elevated l e v e l s of serum TNF. I n active CD o r UC, a greater fraction o f p a t i e n t s studied had s i g n i f i c a n t l y increased serum TNF levels (3/3 f o r CD and S / l 1 f o r UC). Production of TNF by peripheral b l o o d m o n o c y t e s when stimulated b y lipopolysaccharide was a l s o increased in t h e s e patients and correlated with t h e i r serum TNF levels. These rusults suggest that TNF may have s o m e p a t h o e t i o l o g i c a l meaning i n IBD.
INTRODUCTION Many immunological disturbances are associated with (1-6).
Increased h e l p e r T c e l l s (CD4 p o s i t i v e )
IBD
and decreased
suppressor T c e l l s (CDS p o s i t i v e ) populations in t h e peripheral circulation were found frequently i n UC (1).
In addition, enhanced
expression o f h u m a n leukocyte antigen (HLA) was recently correlated with
IBD
(2).
Furthermore,
451 Copyright @ 1992 by Marcel Dekker, Inc.
immune
cyt o k i n e s
including
MAEDA ET AL.
452 TABLE 1 Patients Profiles
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
uc
CD
Sex
28 M13, F15
5 M4, F1
Age(y r) Disease activity
19-42 active remission
23-33 active remission
3 2
Extent of lesion
total colitis left-sided colitis proc titis
ileocolitis colitis
3 2
No of cases
11 17 7 15
6
interleukin-1 (IL-1) (3), interferon (IFN) (4, 5 ) and TNFare assumed to be involved in t h e hyperimmune s t a t e of IBD. I n the present p a p e r , we describe serum TNF levels and TNF production by circulating monocytcs i n p a t i c n t s w i t h IBD.
PATIENTS AND METHODS Human Serum Samoles We studied 3 3 p a t i e n t s ( 1 6 male and 1 7 female, aged 1 9 t o 46 years) w i t h IBD.
The p a t i e n t s had a h i s t o l o g i c a l l y
confirmed
diagnosis of IBD. The p a t h o l o g y corresponded t o UC i n 2 8 cases (active stage cases (active
:
11 cases, remission s t a g e : 3, remission
:
2) (TABLE1).
Discase c o n t r o l s were as follows polyp ( n = 3 ) , ischemic
17 cases), and CD i n 5
c o l i t i s (n=2),
c o l o n cancer ( n = l 9 ) , colon infectious c o l i t i s
(n=8),
pneumonia ( n = 4 ) , c h o l e c y s t i t i s (n=3), and h e p a t i t i s ( n = 3 ) . Twenty healthy age-matchcd laboratory workers s e r v e d a s normal c o n t r o l s .
453
TUMOR N E C R O S I S FACTOR I N BOWEL D I S E A S E
Serum samples were inactivated by heating a t 56C for 3 0 m i n , and
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
were stored a t -70 “c un ti1 assayed. Recomb in an t Human TNF Recombinant human TNF (rhTNF), produced in Escherichia coli and purified (99.9%) was generously provided by Asahi Chemical Ind. C o . , Ltd. (Tokyo, Japan) (7). The rhTNF had a s p e c i f i c a c t i v i t y of 2.37 X l o 6 units (U)/mg protein a s determined b y i t s c y t o t o x i c activity a g a i n s t TNF s e n s i t i v e L-M (mouse tumorigenic f i b r o b l a s t cells, ATCC CLL 137, American Type Culture Collection, R o c k v i l l e ,
MD.) (8). A TNF activity unit (U) was defined as t h e amount of TNF g i v i n g 50% L c e l l survival as determined by dye-uptake (7). ELISA (Enzyme Linked Immunosorbent Assay) Serum TNF levels were measured by a double antibody enzyme linked immunosorbent assay (ELISA, Asahi Chemical Ind. C o . , Ltd.) (9). Microtiter plates (Dynatech, Immulon 11, 96 wells) were coated with rabbit anti-rhTNF IgG by o v e r n i g h t incubation with a n t i b o d y at 4°C. The plates were then washed 5 times with p h o s p h a t e buffered s a l i n e (PBS) c o n t a i n i n g 0 . 2 % Triton X-100. Next, aliquots o f serum from p a t i e n t s were added to t h e wells. Serially diluted rhTNF i n dog plasma was included to establish a standard curve. The p l a t e s were then incubated with serum samples at 4°C for 2 4 h . After washing w i t h PBS c o n t a i n i n g 0 . 2 % Triton X-100, peroxidase conjugated
anti-TNF a n t i b o d y was added. The plates were then incubated for 6 h at r o o m temperature. After washing, they were incubated with p-n itro-phen ylphosphat e substrate a t room temperature for 1 O m i n . The absorbance of product was measured a t 492nm using an ELISA reader (SLT-Lab Instruments, Austria). TNF levels were detected i n a dose dependent manner a t concentrations between 0 . 2 and 2.0 U/ml. The determinations were made in triplicate. TNF i n
serum was
The detection limit of
about 0.2U/ml corresponding t o
concentration of 10Opg/ml by t h i s assay.
a rhTNF
454
MAEDA ET A L .
Isolation a n d c u l t u r e o f Human Peripheral Blood Monocytes Mo n o n ucl ear cell s were i so I a ted from h ep ari ni zed p eri ph er a1 Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
venous b l o o d (1OU heparin/ml b l o o d ) b y Ficoll-Isopaque gradient density centrifugation. The m o n o c y t e fraction was isolated using macrophage separation p l a t e (MSP-P, Otsuka Assay Laboratories, Osaka, Japan).
The monocyte concentration was adjusted t o 5 X
106/ml i n Eagle's minimal e s s e n t i a l medium (EMEM) with
10%
h e a l - i n a c t i v a t e d f e t a l b o v i n e serum (FBS, Flow L a b . , Australia), 1 0 0 U/ml of p e n i c i l l i n , 1OOp g/ml of streptomycin and 10mM HEPES
(pH7.2). These c e l l s were t h e n incubated for 20h in t h e presence o r absence o f 1 0 p g/ml lipopolysaccharide (DIFCO, LPS B , E. c o l i , 01 27
B8) a t 37°C and 5 % CO, in humidified a i r . The supernatants
were collected after 2 0 h a s conditioned media, filtered through a 0 . 2 2 p m m i l l i p o r e filter,and kept frozen at -20°C until assayed.
Bioasaay for TNF
To measure t h e TNF a c t i v i t y i n a b i o a s s a y , standardized serum samples and conditioned media of monocytes from IBD patients were incubated with Actinomycin D (final concentration l p g/ml) treated L-M c e l l s at 37°C for 48h in 5 % CO,.
C y t o t o x i c activity was t h e n
a s s e s s e d b y the dye uptake method ( 1 0 , l l ) .
RESULTS Serum Levels o f TNF i n Patients with IBD --TheserumTNFlevels of all20 healthy subjects were below detection l i m i t . W i t h IBD, on t h e o t h e r hand, p o s i t i v e e l e v a t i o n s were observed i n 8 0 % (4/5) of p a t i e n t s with CD and 31% ( 9 / 2 8 ) of patients with UC (FIG. 1 ) . The percentages of p o s i t i v e e l e v a t i o n s in p a t i e n t s with t h e a c t i v e stage were as much as 1 0 0 % ( 3 / 3 ) for CD and 7 2 . 7 % (8/1 1) for UC. I n t h e s e i n s t a n c e s , serum levels of TNF were high at 0 . 4 2 - 0 . 8 6 U/ml for C D a n d 0 . 2 - 2 . 1 2 U/ml f o r UC.
455
TUMOR N E C R O S I S FACTOR I N BOWEL D I S E A S E
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
A 2.12
A A
_ _ _ _ *_ _ _ _ - _ _ _ _ _ - _ 0.0.
Healthy Crohn's Ulcerative Infectious Colon Other Colon Other Control Disease Colitis Colitis Cancer Disease Inflammatory (n=20) (n=5) (n=28) (n=8) (n=19) (n=5) Disease (n=10)
FIG. 1. Serum TNF concentration i n p a t i e n t s with c o l o n diseases. In I B D p a t i e n t s , A a n d A represented a c t i v e and remitted disease, respectively. Serum samples listed under o t h e r colon diseases were taken from p a t i n t s with ischemic c o l i t i s (m) and p o l y p s
(a).
As f o r o t h e r diseases o f t h e c o l o n , increased serum TNF l e v e l s
were seen i n 26.7% ( 3 / 8 ) p a t i e n t s with infectious c o l i t i s , 9 . 5 % (2/19) o f p a t i e n t s with c o l o n cancer,and 1 0 0 % (2/2) of p a t i e n t s with ischemic c o l i t i s a s shown i n other colon disease group o f F i g . 1 , however, serum TNF levels i n p a t i e n t s with these diseases were lower than t h o s e in p a t i e n t s with a c t i v e IBD.
The serum TNF in
these c o n t r o l s was n o more than 0.27 U/ml (patients with colon cancer had t h e highest levels). The three p a t i e n t s with c o l o n p o l y p s showed no p o s i t i v e result. Inflammatory diseases i n s i t e s other than t h e large i n t e s t i n e ( h e p a t i t i s , 3 cases; c h o l e c y s t i t i s , 3 c a s e s ; pneumonia, 4 c a s e s ) , a
MAEDA ET AL.
456 2.12
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
0.9-
>-
It7
(pre ope)
0.80.7-
53
.- 0.6Y
P
0.5-
u C
2 z k
0.4-
IJ.
0.3-
therapy
therapy
FIG. 2. Serum TNF concentration i n IBD p a t i e n t s before and after therapy. I n IBD p a t i e n t s , 0 a n d 0 represented UC and CD, respectively .
p o s i t i v e e l e v a t i o n s were obtained only in two cases o f pneumonia; the serum TNF levels i n t h e s e cases were l o w a t 0 . 2 6 U/ml and 0 . 3 1 U/ml compared to t h o s e in cases o f UC and CD. Relationship between P a t h o l o g i c Features o f IBD and Serum TNF Serum l e v e l s o f TNF were measured i n t h r e e CD p a t i e n t s and s i x UC p a t i e n t s before and a f t e r treatment. Of t h e p a t i e n t s with UC, o n e case received corticosteroid, a n o t h e r underwent surgery, and t h e other four cases received sulfasalazine. Of t h e p a t i e n t s with CD, o n e case received corticosteroid, a n o t h e r sulfasalazine o r elemental diet therapy. Regardless o f t h e treatment regimen, serum TNF decreased below t h e detection limit(O.2U/ml) i n b o t h CD and UC (FIG. 2), and correlated well with r e m i s s i o n . Furthermore, i n o n e C D patient between serum TNF levels,
there was g o o d correlation
C-react v e p r o t e i n ,
(Crohn's disease a c t i v e index) (FIG. 3 )
and the CDAI
- -6oi\ y-yF:200
457
TUMOR N E C R O S I S FACTOR I N BOWEL D I S E A S E ESR
Immunopharmacology and Immunotoxicology Downloaded from informahealthcare.com by York University Libraries on 12/28/14 For personal use only.
SERUM TNF (Ufml)
70
CDAl
(Ihr)
50
ESR
40
.
10
.
0
J
150
0.6
Q
CRP
0 0.
--
Da :e
lj ar
.inn
0.54 ...
.11
-Eo4++NF
2/27
I
50
---o
0
