- Email: [email protected]
The relationship between pneumocyte reactivity and the outcome of experimental streptococcal pneumonia
35
EXPERIMENTAL PATHOLOGY MACROSCOPIC AN11 MICROSCOFIC A N A L Y S I S DT ETllANOL A N D N I C O I ' L N I ? I)AMAl:E '7'0 RAT GASTRIC MIICOSA AFTER TREATMLNT WlTIi MISOPROSTIL AND OMEPRAZOIX W , M . Ilui, R . W . Chen, S.K. L a m , Department o f Fnthnloqi. all4 Medicine, l l n i v c r s j t y o r Hong Konq
*L.J.,
W e compared Lhe p r o t e c t i v e P f f e c t s of m i s o p r o s t o l , a p r o s t a g l a n d i r i E l d e r i v a t i v e w i t h b o t h a c i d - i n h i b i t o r y and c y t o p r o t e c t i v e a c t i o n s , t o omeprazole which p o s s e s s e s "otent acid-inhibiLory a c t i v i t y only on t h e i n j u r y t o r a t g a s t r i c mucosa induced by e t h a n o l a l o n e , and e t h a n o l combined w i t h n i c o t i n e , which was give11 t o r a t s 25lJg/ml t a p H$ f o r t e n days b e f o r e experiment. Assessment of macros c o p i c and m i c r o s c o p i c mucosal damage were made i n r a t s t r e a t e d w i t h e t h a n o l and n i c o t i n e a l o n e , o r i n combination w i t h v a r i e d dosrs of M i s o p r o s t o l , and Omeprazole t o p i c a l l y Tlie u l c e r index was measured by t h e t o t a l a r e a o f macros c o p i c hpmorrhnqic n e c r o s i s i n t h e g l a n d u l a r stomach. Microscopic examination of t h e t i s s u e t a k e n from t h e nonn e c r o t i c l e s i o n a r e a s o f , e a c h r a t stomach was c a r r i e d o u t i n o r d e r t o d e t e r m i n e t h e e x t e n t and d e p t h o f m u c o s a l damage. The d e p t h oE mu'cosal damage was c l a s s i f i e d a s 0-damage; I-damage, 11-damat~e and 111-damage based on standardized c r i t e r i a . In r a t s challenged with ethanol a l o n e , t h e u l c e r i n d e x , t h e t o t a l a r e a of damaged g a s t r i c mucosa, and I11 damage i n t h e n o n - n e c r o t i c l e s i o n a r e a of stomach were s i g n i f i c a n t l y lower i n r a t s t r e a t e d w i t h a l l d o s e s of rnisoprostol and high d o s e s of Omeprazol? vs l o w d o s e s of Omeprazole and c o n t r o l ( y C O . 0 5 ) . In rat.s c h a l l e n g e d w i t h e t h a n o l i n combination w i t h c h r o n i c n i c o t i n e , t h e u l c e r i n d e x , t o t a l a r e a of damaged mucosa and 1 1 1 damaqe i n n o n - n e c r o t i c l e s i o n a r e a s were s i g n i f i c a n t l y lower i n r a t s t r e a t e d w i t h a l l d o s e s of misoprostol v s a l l d o s e s of Omeprazole and c o n t r o l (p
Presenter:
Dr. J. 110,
Department of P a t h o l o g y , H K U
THE RELATIONSHIP BETWEEN PNEUMOCYTE R E A C T I V I T Y AND T H E OUTCOME OF EXPERIMENTAL STREPTOCOCCAL PNEUMONIA GC Rhodes*, JW T a p s a l l , AWJ Lykke School o f Pathology, U n i v e r s i t y o f NSW, Kensington 2033
Two models o f b a c t e r i a l pneumonia i n SPF A u s t r a l i a n
A1 b i n o W i s t a r r a t s have been developed and c h a r a c t e r i s e d
i n o r d e r t h a t a l v e o l a r e p i t h e l i a l (pneumocyte) r e a c t i v i t y may be s t u d i e d d u r i n g t h e e v o l u t i o n o f t h i s d i s e a s e . I n t h e f i r s t model, i n which pneumonia i s induced by a s t r a i n o f Streptococcus sanguis, r e s o l u t i o n o f the pneumonic l e s i o n i s a s s o c i a t e d w i t h r a p i d r e p a i r o f damaged a l v e o l a r e p i t h e l i u m mediated by h y p e r p l a s i a o f t y p e 2 pneumocytes and t r a n s f o r m a t i o n o f a p r o p o r t i o n o f daughter c e l l s i n t o t y p e 1 pneumocytes'. I n t h e second model, i n which pneumonia i s induced by a s t r a i n o f S t r e p t o c o c c u s pneumoniae t y p e 25, f a i l u r e o f r e s o l u t i o n i s a s s o c i a t e d w i t h damage t o t r a n s f o r m i n g t y p e 2 pneumoc y t e s and subsequent i n h i b i t i o n o f t h e normal a l v e o l a r e p i t h e l i a l r e p a i r process'. Using immunohistochemistry t o i d e n t i f y t y p e 2 pneumoc y t e s and a comparative morphometric a n a l y s i s , we have c o n f i r m e d s i g n i f i c a n t d i f f e r e n c e s between t h e two models i n t h e p a t t e r n o f t h i s c e l l ' s response. These f i n d i n g s have suggested t h a t t h e t y p e 2 pneumocyte's response t o an i n j u r i o u s s t i m u l u s may be o f fundamental importance i n d e t e r m i n i n g t h e u l t i m a t e outcome o f t h e a c u t e i n j u r y .
1.
Rhodes, GC, T a p s a l l JW, .Lykke AWJ. A l v e o l a r e p i t h e l i a l response i n e x p e r i m e n t a l s t r e p t o c o c c a l pneumonia. J P a t h o l (1989) 157: 347-357.
2.
Rhodes GC, Lykke AWJ, T a p s a l l JW, S m i t h LW. Abnormal a l v e o l a r e p i t h e l i a l r e p a i r associated w i t h f a i l u r e o f r e s o l u t i o n i n e x p e r i m e n t a l s t r e p t o c o c c a l pneumonia J Pathol ( i n press)
THE INIITBITORY FFFFCT OF APOLIPOPROTEINS I N IIDL ON FXI'W TMEXrA L ATHEROSCLEROSIS I N RABBITS Bi-fan3 W_anxzoiT l.C_h_eF-B&o:shengt Xia Ren-yi. S_he-Ming:pygfr Department of Pathology, I n s t i t u t e of B a s i c Medical S c i e n c e s , Chinese Academy of Medical S c i e n c e s , R e i j i n R 100730 China
Ran ~.
According t o d a t a obtained from e p i d e m i o l o g i c a l and exp e r i m e n t a l s u r v e y s , serum HDL l e v e l is known t o be corr e l a t e d r e v e r s e l y with t h e i n c i d e n c e of a t h e r o a c l e r o e i e . The main a p o l i p o p r o t e i n s i n HDL are known t o be apo A , A and A Apolipoprotpine (mainly apo A ) were i s o l a t e d from about 100 l i t e r e of r a b b i t ' a e e m f o r azd s t u d y i n g t h e i n h i b i t o r y e f f e c t o n a t h e r o m a t o u s plaque development i n r a b b i t s . Data i r d i c a t e d t h a t l i p i d c o n t e n t i n a o r t i c i n t i m a ( t a b l e below shows d a t a of experiment 1 ) . area of n t h e r o m a t o u ~l e e i a n involved, l i p i d d e p o s i t i o n i n i n t i m a m o r p h o l o g i c a l l y and t h e s e v e r i t y of atheromatous l e s i o n s involved were much lower i n a n i m a l s o f t h e e x p e r i mental group (with c h o l e s t e r o l 6 days/wk for 4-12 weeks) a f t e r a d m i n s t r a t i o n i n t r a v e n o u s l y of a p o l i p o p r o t e i n s i s o l a t e d f r a n HDL for 8 t o 12 weeks t h a n t h o e e of a n i m a l s of t h e c h o l e s t e r o l c o n t r o l group i n two s u c c e s s i v e e x p e r i ments. Data of t h e s e e x p e r i m e n t s c l a r i f i e d t h a t a p o l i p o p r o t e i n s (mainly apo A ) i n llDL a r e t h e main f a c t o r s ac1 t i n g on t h i s i n h i b i t o r y e f f e c t .
purified
~
groups
l i p i d s i n i n t i m a (mg/pm wet w t . )
chol. c o n t r o l (6 a n i m a l s ) experimental
(6)
blank c o n t r o l
(6)
T
a
m
n
t t l . chol. 16.5 4.8
cho1,est. 8.9 5 2.0
6.7 5 9.4
3.8 5 5.6 ([email protected]) 0.6 + 0.2 (P'ij.01)
(pt0.05) 1.7 t 0.1 (P
m Molecular
B i G V
f r e e chol.
-
7.6 5 2.9
2.9 2 7.8
(~'0.05) 1.2 + 0.1 (P
Biochemistry
-
ADJUVANT ARTHRITIS AND PERITONEAL MACROPHAGE ACTIVATION. T.R. Southwood*, R.E. P e t t y . J.B. Bradley**. D i v i s i o n o f Rheumatology, Department o f P a e d i a t r i c s , U n i v e r s i t y o f B r i t i s h Columbia, Vancouver, CANADA, and **Department o f C l i n i c a l Immunology, F l i n d e r s Medical Centre, South AUSTRALIA. The c e l l u l a r e v e n t s a s s o c i a t e d w i t h complete Freund's a d j u v a n t (CFA)-induced a r t h r i t i s (AA), a r a t model o f human a r t h r i t i s , a r e u n c l e a r . Our o b j e c t i v e was t o examine t h e r e l a t i o n s h i p between p e r i t o n e a l macrophage (PMP) a c t i v a t i o n and t h e a r t h r i t i c response t o a d j u v a n t i n Sprague-Dawley r a t s . P e r i t o n e a l c e l l s were o b t a i n e d by l a v a g e b e f o r e (day 0 ) and a f t e r CFA (day 21), and t h e PMP p o p u l a t i o n was d e f i n e d by c e l l s i z e and g r a n u l a r i t y u s i n g f l o w c y t o m e t r y . 75% t o 90% o f c e l l s f r o m t h i s p o p u l a t i o n were p h a g o c y t l c , adherent and NSE p o s i t i v e . A c t i v a t e d PMP (aPMP) (PMP e x p r e s s i n g MHC c l a s s I 1 (RT1.D) a n t i g e n s ) , were measured by i n d i r e c t immunof l u o r e s c e n c e u s i n g t h e monoclonal a n t i b o d y 0x17. Severe AA (n-14) was a s s o c i a t e d w i t h a s i g n i f i c a n t i n c r e a s e i n mean % aPMP (day 0:day 21 10.7:41.2, P-.0001). Rats w i t h m i l d AA (n-6) had a s m a l l e r i n c r e a s e i n aPMP (day 0:day 21 19.4:28.0, P - . 2 ) than d i d r a t s w i t h severe AA (P-.Ol). M i l d AA was a s s o c i a t e d w i t h a s l i g h t l y h i g h e r aPMP a t day 0 than severe AA (P-.3). I n t r a p e r i t o n e a l i n j e c t i o n o f c u l t u r e s u p e r n a t a n t from Con A s t i m u l a t e d spleen c e l l s i n c r e a s e d aPMP (45%) compared t o s a l i n e - i n j e c t e d r a t s (aPMP-18%), (P-.016). Twelve days a f t e r CFA, o n l y 1/5 r a t s w i t h h i g h i n i t i a l aPMP (meant2SD) had developed AA, whereas 6/7 r a t s w i t h normal aPMP had AA (P=.OZ). I n c o n c l u s i o n , these r e s u l t s suggest aPMP may i n c r e a s e i n response t o t h e i n f l a m m a t i o n a s s o c i a t e d w i t h severe AA, b u t r a t s w i t h h i g h aPMP b e f o r e CFA i s g i v e n may be p r o t e c t e d a g a i n s t AA.
EXPERIMENTAL PATHOLOGY MACROSCOPIC AN11 MICROSCOFIC A N A L Y S I S DT ETllANOL A N D N I C O I ' L N I ? I)AMAl:E '7'0 RAT GASTRIC MIICOSA AFTER TREATMLNT WlTIi MISOPROSTIL AND OMEPRAZOIX W , M . Ilui, R . W . Chen, S.K. L a m , Department o f Fnthnloqi. all4 Medicine, l l n i v c r s j t y o r Hong Konq
*L.J.,
W e compared Lhe p r o t e c t i v e P f f e c t s of m i s o p r o s t o l , a p r o s t a g l a n d i r i E l d e r i v a t i v e w i t h b o t h a c i d - i n h i b i t o r y and c y t o p r o t e c t i v e a c t i o n s , t o omeprazole which p o s s e s s e s "otent acid-inhibiLory a c t i v i t y only on t h e i n j u r y t o r a t g a s t r i c mucosa induced by e t h a n o l a l o n e , and e t h a n o l combined w i t h n i c o t i n e , which was give11 t o r a t s 25lJg/ml t a p H$ f o r t e n days b e f o r e experiment. Assessment of macros c o p i c and m i c r o s c o p i c mucosal damage were made i n r a t s t r e a t e d w i t h e t h a n o l and n i c o t i n e a l o n e , o r i n combination w i t h v a r i e d dosrs of M i s o p r o s t o l , and Omeprazole t o p i c a l l y Tlie u l c e r index was measured by t h e t o t a l a r e a o f macros c o p i c hpmorrhnqic n e c r o s i s i n t h e g l a n d u l a r stomach. Microscopic examination of t h e t i s s u e t a k e n from t h e nonn e c r o t i c l e s i o n a r e a s o f , e a c h r a t stomach was c a r r i e d o u t i n o r d e r t o d e t e r m i n e t h e e x t e n t and d e p t h o f m u c o s a l damage. The d e p t h oE mu'cosal damage was c l a s s i f i e d a s 0-damage; I-damage, 11-damat~e and 111-damage based on standardized c r i t e r i a . In r a t s challenged with ethanol a l o n e , t h e u l c e r i n d e x , t h e t o t a l a r e a of damaged g a s t r i c mucosa, and I11 damage i n t h e n o n - n e c r o t i c l e s i o n a r e a of stomach were s i g n i f i c a n t l y lower i n r a t s t r e a t e d w i t h a l l d o s e s of rnisoprostol and high d o s e s of Omeprazol? vs l o w d o s e s of Omeprazole and c o n t r o l ( y C O . 0 5 ) . In rat.s c h a l l e n g e d w i t h e t h a n o l i n combination w i t h c h r o n i c n i c o t i n e , t h e u l c e r i n d e x , t o t a l a r e a of damaged mucosa and 1 1 1 damaqe i n n o n - n e c r o t i c l e s i o n a r e a s were s i g n i f i c a n t l y lower i n r a t s t r e a t e d w i t h a l l d o s e s of misoprostol v s a l l d o s e s of Omeprazole and c o n t r o l (p
Presenter:
Dr. J. 110,
Department of P a t h o l o g y , H K U
THE RELATIONSHIP BETWEEN PNEUMOCYTE R E A C T I V I T Y AND T H E OUTCOME OF EXPERIMENTAL STREPTOCOCCAL PNEUMONIA GC Rhodes*, JW T a p s a l l , AWJ Lykke School o f Pathology, U n i v e r s i t y o f NSW, Kensington 2033
Two models o f b a c t e r i a l pneumonia i n SPF A u s t r a l i a n
A1 b i n o W i s t a r r a t s have been developed and c h a r a c t e r i s e d
i n o r d e r t h a t a l v e o l a r e p i t h e l i a l (pneumocyte) r e a c t i v i t y may be s t u d i e d d u r i n g t h e e v o l u t i o n o f t h i s d i s e a s e . I n t h e f i r s t model, i n which pneumonia i s induced by a s t r a i n o f Streptococcus sanguis, r e s o l u t i o n o f the pneumonic l e s i o n i s a s s o c i a t e d w i t h r a p i d r e p a i r o f damaged a l v e o l a r e p i t h e l i u m mediated by h y p e r p l a s i a o f t y p e 2 pneumocytes and t r a n s f o r m a t i o n o f a p r o p o r t i o n o f daughter c e l l s i n t o t y p e 1 pneumocytes'. I n t h e second model, i n which pneumonia i s induced by a s t r a i n o f S t r e p t o c o c c u s pneumoniae t y p e 25, f a i l u r e o f r e s o l u t i o n i s a s s o c i a t e d w i t h damage t o t r a n s f o r m i n g t y p e 2 pneumoc y t e s and subsequent i n h i b i t i o n o f t h e normal a l v e o l a r e p i t h e l i a l r e p a i r process'. Using immunohistochemistry t o i d e n t i f y t y p e 2 pneumoc y t e s and a comparative morphometric a n a l y s i s , we have c o n f i r m e d s i g n i f i c a n t d i f f e r e n c e s between t h e two models i n t h e p a t t e r n o f t h i s c e l l ' s response. These f i n d i n g s have suggested t h a t t h e t y p e 2 pneumocyte's response t o an i n j u r i o u s s t i m u l u s may be o f fundamental importance i n d e t e r m i n i n g t h e u l t i m a t e outcome o f t h e a c u t e i n j u r y .
1.
Rhodes, GC, T a p s a l l JW, .Lykke AWJ. A l v e o l a r e p i t h e l i a l response i n e x p e r i m e n t a l s t r e p t o c o c c a l pneumonia. J P a t h o l (1989) 157: 347-357.
2.
Rhodes GC, Lykke AWJ, T a p s a l l JW, S m i t h LW. Abnormal a l v e o l a r e p i t h e l i a l r e p a i r associated w i t h f a i l u r e o f r e s o l u t i o n i n e x p e r i m e n t a l s t r e p t o c o c c a l pneumonia J Pathol ( i n press)
THE INIITBITORY FFFFCT OF APOLIPOPROTEINS I N IIDL ON FXI'W TMEXrA L ATHEROSCLEROSIS I N RABBITS Bi-fan3 W_anxzoiT l.C_h_eF-B&o:shengt Xia Ren-yi. S_he-Ming:pygfr Department of Pathology, I n s t i t u t e of B a s i c Medical S c i e n c e s , Chinese Academy of Medical S c i e n c e s , R e i j i n R 100730 China
Ran ~.
According t o d a t a obtained from e p i d e m i o l o g i c a l and exp e r i m e n t a l s u r v e y s , serum HDL l e v e l is known t o be corr e l a t e d r e v e r s e l y with t h e i n c i d e n c e of a t h e r o a c l e r o e i e . The main a p o l i p o p r o t e i n s i n HDL are known t o be apo A , A and A Apolipoprotpine (mainly apo A ) were i s o l a t e d from about 100 l i t e r e of r a b b i t ' a e e m f o r azd s t u d y i n g t h e i n h i b i t o r y e f f e c t o n a t h e r o m a t o u s plaque development i n r a b b i t s . Data i r d i c a t e d t h a t l i p i d c o n t e n t i n a o r t i c i n t i m a ( t a b l e below shows d a t a of experiment 1 ) . area of n t h e r o m a t o u ~l e e i a n involved, l i p i d d e p o s i t i o n i n i n t i m a m o r p h o l o g i c a l l y and t h e s e v e r i t y of atheromatous l e s i o n s involved were much lower i n a n i m a l s o f t h e e x p e r i mental group (with c h o l e s t e r o l 6 days/wk for 4-12 weeks) a f t e r a d m i n s t r a t i o n i n t r a v e n o u s l y of a p o l i p o p r o t e i n s i s o l a t e d f r a n HDL for 8 t o 12 weeks t h a n t h o e e of a n i m a l s of t h e c h o l e s t e r o l c o n t r o l group i n two s u c c e s s i v e e x p e r i ments. Data of t h e s e e x p e r i m e n t s c l a r i f i e d t h a t a p o l i p o p r o t e i n s (mainly apo A ) i n llDL a r e t h e main f a c t o r s ac1 t i n g on t h i s i n h i b i t o r y e f f e c t .
purified
~
groups
l i p i d s i n i n t i m a (mg/pm wet w t . )
chol. c o n t r o l (6 a n i m a l s ) experimental
(6)
blank c o n t r o l
(6)
T
a
m
n
t t l . chol. 16.5 4.8
cho1,est. 8.9 5 2.0
6.7 5 9.4
3.8 5 5.6 ([email protected]) 0.6 + 0.2 (P'ij.01)
(pt0.05) 1.7 t 0.1 (P
m Molecular
B i G V
f r e e chol.
-
7.6 5 2.9
2.9 2 7.8
(~'0.05) 1.2 + 0.1 (P
Biochemistry
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ADJUVANT ARTHRITIS AND PERITONEAL MACROPHAGE ACTIVATION. T.R. Southwood*, R.E. P e t t y . J.B. Bradley**. D i v i s i o n o f Rheumatology, Department o f P a e d i a t r i c s , U n i v e r s i t y o f B r i t i s h Columbia, Vancouver, CANADA, and **Department o f C l i n i c a l Immunology, F l i n d e r s Medical Centre, South AUSTRALIA. The c e l l u l a r e v e n t s a s s o c i a t e d w i t h complete Freund's a d j u v a n t (CFA)-induced a r t h r i t i s (AA), a r a t model o f human a r t h r i t i s , a r e u n c l e a r . Our o b j e c t i v e was t o examine t h e r e l a t i o n s h i p between p e r i t o n e a l macrophage (PMP) a c t i v a t i o n and t h e a r t h r i t i c response t o a d j u v a n t i n Sprague-Dawley r a t s . P e r i t o n e a l c e l l s were o b t a i n e d by l a v a g e b e f o r e (day 0 ) and a f t e r CFA (day 21), and t h e PMP p o p u l a t i o n was d e f i n e d by c e l l s i z e and g r a n u l a r i t y u s i n g f l o w c y t o m e t r y . 75% t o 90% o f c e l l s f r o m t h i s p o p u l a t i o n were p h a g o c y t l c , adherent and NSE p o s i t i v e . A c t i v a t e d PMP (aPMP) (PMP e x p r e s s i n g MHC c l a s s I 1 (RT1.D) a n t i g e n s ) , were measured by i n d i r e c t immunof l u o r e s c e n c e u s i n g t h e monoclonal a n t i b o d y 0x17. Severe AA (n-14) was a s s o c i a t e d w i t h a s i g n i f i c a n t i n c r e a s e i n mean % aPMP (day 0:day 21 10.7:41.2, P-.0001). Rats w i t h m i l d AA (n-6) had a s m a l l e r i n c r e a s e i n aPMP (day 0:day 21 19.4:28.0, P - . 2 ) than d i d r a t s w i t h severe AA (P-.Ol). M i l d AA was a s s o c i a t e d w i t h a s l i g h t l y h i g h e r aPMP a t day 0 than severe AA (P-.3). I n t r a p e r i t o n e a l i n j e c t i o n o f c u l t u r e s u p e r n a t a n t from Con A s t i m u l a t e d spleen c e l l s i n c r e a s e d aPMP (45%) compared t o s a l i n e - i n j e c t e d r a t s (aPMP-18%), (P-.016). Twelve days a f t e r CFA, o n l y 1/5 r a t s w i t h h i g h i n i t i a l aPMP (meant2SD) had developed AA, whereas 6/7 r a t s w i t h normal aPMP had AA (P=.OZ). I n c o n c l u s i o n , these r e s u l t s suggest aPMP may i n c r e a s e i n response t o t h e i n f l a m m a t i o n a s s o c i a t e d w i t h severe AA, b u t r a t s w i t h h i g h aPMP b e f o r e CFA i s g i v e n may be p r o t e c t e d a g a i n s t AA.