Current Eye Research
ISSN: 0271-3683 (Print) 1460-2202 (Online) Journal homepage: http://www.tandfonline.com/loi/icey20
Protective effects of anti-glycoprotein D monoclonal antibodies in murine herpetic keratitis Yoshitsugu Inoue, Yuichi Ohashi, Hitoshi Watanabe & Reizo Manabe To cite this article: Yoshitsugu Inoue, Yuichi Ohashi, Hitoshi Watanabe & Reizo Manabe (1992) Protective effects of anti-glycoprotein D monoclonal antibodies in murine herpetic keratitis, Current Eye Research, 11:1, 53-60, DOI: 10.3109/02713689209069167 To link to this article: http://dx.doi.org/10.3109/02713689209069167
Published online: 02 Jul 2009.
Submit your article to this journal
Article views: 2
View related articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=icey20 Download by: [New York University]
Date: 07 April 2016, At: 17:48
Current Eye Research
Volume 1 1 number I 1992, 53-60
Protective effects of anti-glycoprotein D monoclonal antibodies in murine herpetic keratitis Yoshitsugu Inoue, Yuichi Ohashi, Hitoshi Watanabe and Reizo Manabe ~
~~
Downloaded by [New York University] at 17:48 07 April 2016
Department of Ophthalmology, Osaka University Medical School, Fukushima-ku, Osaka 553, Japan
ABSTRACT The p r o t e c t i v e e f f e c t s of p a s s i v e immunization w i t h two k i n d s of a n t i - g l y c o p r o t e i n D (anti-gD) monoclonal a n t i b o d i e s , h a v i n g d i f f e r e n t a n t i v i r a l a c t i v i t i e s , were i n v e s t i g a t e d i n murine h e r p e t i c k e r a t i t i s . One monoclonal a n t i b o d y , d e s i g n a t e d M1, had h i g h v i r u s - n e u t r a l i z i n g a n t i b o d y t i t e r s , a l o n g w i t h u n d e t e c t a b l e l e v e l s of complement-dependent c y t o l y s i s (CDC) and antibody-dependent c e l l u l a r c y t o t o x i c i t y (ADCC); t h e o t h e r , d e s i g n a t e d M12, e x h i b i t e d e x t r e m e l y low t i t e r s o f v i r u s - n e u t r a l i z a t i o n w i t h h i g h l e v e l o f CDC and ADCC. When s y s t e m i c a l l y a d m i n i s t e r e d 24 h o u r s p r i o r t o v i r u s i n o c u l a t i o n t o t h e c o r n e a , b o t h M1 and M12 a l m o s t c o m p l e t e l y p r e v e n t e d t h e development o f s t r o m a l k e r a t i t i s . The p r o t e c t i v e e f f i c a c y of b o t h w a s o b s e r v e d t o b e dose-dependent. P e p s i n - t r e a t e d M1 r e t a i n e d its efficacy i n suppressing stromal k e r a t i t i s , whereas p e p s i n - t r e a t e d M12 d i d n o t . When t h e a d m i n i s t r a t i o n of M1 and M12 w e r e d e l a y e d , b o t h p r o v i d e d s i g n i f i c a n t ( b u t l e s s complete) p r o t e c t i o n , up t o 2 4 h o u r s a f t e r v i r u s inoculation. These r e s u l t s s u g g e s t t h a t b o t h v i r u s n e u t r a l i z a t i o n and CDC/ADCC p l a y a n i m p o r t a n t r o l e i n p r e v e n t i n g v i r u s growth i n t h e c o r n e a l stroma during t h e e a r l y s t a g e of corneal infection.
from t h e i n f e c t e d c o r n e a (1).
A s w e l l , Metcalf
and h i s c o l l e a g u e s h a v e shown t h a t s e v e r a l c l o n e s of monoclonal a n t i b o d i e s , when p a s s i v e l y administered p r i o r t o v i r u s inoculation, can prevent stromal k e r a t i t i s ( 2 , 3 ) .
However, i t
remains u n c l e a r as t o which mechanism of humoral immunity i s r e s p o n s i b l e f o r t h i s p r o t e c t i o n . The p r e s e n t s t u d y w a s u n d e r t a k e n , u s i n g two c l o n e s o f anti-gD monoclonal a n t i b o d i e s having t o t a l l y different anti-viral
a c t i v i t i e s , to elucidate the
r o l e of v i r u s n e u t r a l i z a t i o n and CDC/ADCC i n suppressing corneal stromal i n f e c t i o n .
MATERIALS AND METHODS Cells Vero c e l l s d e r i v e d from A f r i c a n g r e e n monkey kidney and L929 c e l l s d e r i v e d from C3H mice were grown i n E a g l e ’ s minimum e s s e n t i a l medium (MEM) supplemented w i t h 5% f e t a l c a l f serum (FCS). Anima 1s f e m a l e BALB/c mice were
INTRODUCTION
S i x t o e i g h t week-old
A v a r i e t y o f g l y c o p r o t e i n s (gB, gC, gD, gE, gG,
purchased from Oriental-Kobo
gH and g I ) p r e s e n t on t h e HSV v i r i o n e n v e l o p e and
Research conformed t o t h e D e c l a r a t i o n of H e l s i n k i
t h e c e l l membrane i n f e c t e d w i t h HSV a r e t a r g e t s
and t h e Guiding P r i n c i p l e s i n t h e Care and Use of
(Osaka, J a p a n ) .
f o r t h e h o s t immune mechanisms c o m p r i s i n g v i r u s
Animals.
n e u t r a l i z a t i o n , complement-dependent c y t o l y s i s
Virus
(CDC), antibody-dependent
The Kawamura s t r a i n , HSV t y p e 1 , i s o l a t e d from a
cellular cytotoxicity
(ADCC), c y t o t o x i c T lymphocyte (CTL) and d e l a y e d t y p e h y p e r s e n s i t i v i t y (DTH).
It h a s been
patient with recurrent herpetic stromal k e r a t i t i s ,
w a s used t h r o u g h o u t t h e s t u d y ( 4 ) .
The i n f e c t i o u s
g e n e r a l l y h e l d , as a r e s u l t of b a s i c and c l i n i c a l
t i t e r s w e r e determined v i a plaque t i t r a t i o n .
o b s e r v a t i o n , t h a t c e l l u l a r immunity i s o f g r e a t
Anti-gD monoclonal a n t i b o d i e s
importance i n e r a d i c a t i n g h e r p e t i c i n f e c t i o n s .
We have a l r e a d y e s t a b l i s h e d 6 c l o n e s of anti-gD
However, a r e c e n t r e p o r t by Lausch and h i s
monoclonal a n t i b o d i e s ( 4 ) .
a s s o c i a t e s r e v e a l e d t h e s i g n i f i c a n t r o l e of
w e used 2 c l o n e s , d e s i g n a t e d M1 and M12, of t h e s e
antibodies i n e f f i c i e n t l y eliminating the v i r u s
antibodies.
I n t h e present study,
Received on September 13, 1991; accepted on December 9, 1991
@ Oxford University Press
53
Current
Eye Research E;val>i-&ion
o f e f f e c t s ( ~ i Fac;sive immunization w i t h
m n o-i l o n__ s i l___ antibodies - o-. Two
c
titers -~
l o ~ l e so t monoclonal a n t i b o d i e s (MI a n d
The n e u t r a l i z i n g a n t i b o d y t i t e r s were d e t e r m i n e d
M I L ) against g l y c o p r o t e i n D were p r e p a r e d as described pLeviously
((4).
by t h e p l a q u e r e d u c t i o n method i n a manner d e s c r i b e d p r e v i o u s l y ( 4j
UALB/r mice w e r e
.
i n t r a p e t i toned1 Ly i n o c u l a t e d w l t h d i f t e r e n t d o s e s
CDC a s s a y s
of monaLluna1 A n t i b o d i e s (1, 10, 25, 100 pg/mouse)
CDC a s s a y s were p e r f o r m e d as d e s c r i b e d p r e v i o u s l y
24 h o u r 9 b e € , j r r , Lmmediately, 24 h o u r s a f t e r ,
(4).
and rt8 h o i i r s a f t - e r v i r u s i n o c u l a t i o n .
HSV-infected
dnclsthe
bvst
emIi
:he
Ci,rrie3s
wit11
i
ranging
Downloaded by [New York University] at 17:48 07 April 2016
D e t e r m i n a t i o n of s e i - u m - n e u t r a l i z i n g a n t i b o d y
Following
wi t h s o d i u m p e n t o b a r b i t a l ,
> f t-lie m ~ c ew e r p s c a r i E i e d 20 t i m e s
? 6 g d u g e n e e d l e a n d i n f e c t e d w i t h HSV it1
titer,
f r o m 1 . 7 ~ 1 0t o~ 6 . 0 ~ 1 0p ~f u f e d ?yes w e r e examined w i t h
a liantl-slit
itimp i n masked f a s h i o n a s t o s e v e r i t y
ISSN: 0271-3683 (Print) 1460-2202 (Online) Journal homepage: http://www.tandfonline.com/loi/icey20
Protective effects of anti-glycoprotein D monoclonal antibodies in murine herpetic keratitis Yoshitsugu Inoue, Yuichi Ohashi, Hitoshi Watanabe & Reizo Manabe To cite this article: Yoshitsugu Inoue, Yuichi Ohashi, Hitoshi Watanabe & Reizo Manabe (1992) Protective effects of anti-glycoprotein D monoclonal antibodies in murine herpetic keratitis, Current Eye Research, 11:1, 53-60, DOI: 10.3109/02713689209069167 To link to this article: http://dx.doi.org/10.3109/02713689209069167
Published online: 02 Jul 2009.
Submit your article to this journal
Article views: 2
View related articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=icey20 Download by: [New York University]
Date: 07 April 2016, At: 17:48
Current Eye Research
Volume 1 1 number I 1992, 53-60
Protective effects of anti-glycoprotein D monoclonal antibodies in murine herpetic keratitis Yoshitsugu Inoue, Yuichi Ohashi, Hitoshi Watanabe and Reizo Manabe ~
~~
Downloaded by [New York University] at 17:48 07 April 2016
Department of Ophthalmology, Osaka University Medical School, Fukushima-ku, Osaka 553, Japan
ABSTRACT The p r o t e c t i v e e f f e c t s of p a s s i v e immunization w i t h two k i n d s of a n t i - g l y c o p r o t e i n D (anti-gD) monoclonal a n t i b o d i e s , h a v i n g d i f f e r e n t a n t i v i r a l a c t i v i t i e s , were i n v e s t i g a t e d i n murine h e r p e t i c k e r a t i t i s . One monoclonal a n t i b o d y , d e s i g n a t e d M1, had h i g h v i r u s - n e u t r a l i z i n g a n t i b o d y t i t e r s , a l o n g w i t h u n d e t e c t a b l e l e v e l s of complement-dependent c y t o l y s i s (CDC) and antibody-dependent c e l l u l a r c y t o t o x i c i t y (ADCC); t h e o t h e r , d e s i g n a t e d M12, e x h i b i t e d e x t r e m e l y low t i t e r s o f v i r u s - n e u t r a l i z a t i o n w i t h h i g h l e v e l o f CDC and ADCC. When s y s t e m i c a l l y a d m i n i s t e r e d 24 h o u r s p r i o r t o v i r u s i n o c u l a t i o n t o t h e c o r n e a , b o t h M1 and M12 a l m o s t c o m p l e t e l y p r e v e n t e d t h e development o f s t r o m a l k e r a t i t i s . The p r o t e c t i v e e f f i c a c y of b o t h w a s o b s e r v e d t o b e dose-dependent. P e p s i n - t r e a t e d M1 r e t a i n e d its efficacy i n suppressing stromal k e r a t i t i s , whereas p e p s i n - t r e a t e d M12 d i d n o t . When t h e a d m i n i s t r a t i o n of M1 and M12 w e r e d e l a y e d , b o t h p r o v i d e d s i g n i f i c a n t ( b u t l e s s complete) p r o t e c t i o n , up t o 2 4 h o u r s a f t e r v i r u s inoculation. These r e s u l t s s u g g e s t t h a t b o t h v i r u s n e u t r a l i z a t i o n and CDC/ADCC p l a y a n i m p o r t a n t r o l e i n p r e v e n t i n g v i r u s growth i n t h e c o r n e a l stroma during t h e e a r l y s t a g e of corneal infection.
from t h e i n f e c t e d c o r n e a (1).
A s w e l l , Metcalf
and h i s c o l l e a g u e s h a v e shown t h a t s e v e r a l c l o n e s of monoclonal a n t i b o d i e s , when p a s s i v e l y administered p r i o r t o v i r u s inoculation, can prevent stromal k e r a t i t i s ( 2 , 3 ) .
However, i t
remains u n c l e a r as t o which mechanism of humoral immunity i s r e s p o n s i b l e f o r t h i s p r o t e c t i o n . The p r e s e n t s t u d y w a s u n d e r t a k e n , u s i n g two c l o n e s o f anti-gD monoclonal a n t i b o d i e s having t o t a l l y different anti-viral
a c t i v i t i e s , to elucidate the
r o l e of v i r u s n e u t r a l i z a t i o n and CDC/ADCC i n suppressing corneal stromal i n f e c t i o n .
MATERIALS AND METHODS Cells Vero c e l l s d e r i v e d from A f r i c a n g r e e n monkey kidney and L929 c e l l s d e r i v e d from C3H mice were grown i n E a g l e ’ s minimum e s s e n t i a l medium (MEM) supplemented w i t h 5% f e t a l c a l f serum (FCS). Anima 1s f e m a l e BALB/c mice were
INTRODUCTION
S i x t o e i g h t week-old
A v a r i e t y o f g l y c o p r o t e i n s (gB, gC, gD, gE, gG,
purchased from Oriental-Kobo
gH and g I ) p r e s e n t on t h e HSV v i r i o n e n v e l o p e and
Research conformed t o t h e D e c l a r a t i o n of H e l s i n k i
t h e c e l l membrane i n f e c t e d w i t h HSV a r e t a r g e t s
and t h e Guiding P r i n c i p l e s i n t h e Care and Use of
(Osaka, J a p a n ) .
f o r t h e h o s t immune mechanisms c o m p r i s i n g v i r u s
Animals.
n e u t r a l i z a t i o n , complement-dependent c y t o l y s i s
Virus
(CDC), antibody-dependent
The Kawamura s t r a i n , HSV t y p e 1 , i s o l a t e d from a
cellular cytotoxicity
(ADCC), c y t o t o x i c T lymphocyte (CTL) and d e l a y e d t y p e h y p e r s e n s i t i v i t y (DTH).
It h a s been
patient with recurrent herpetic stromal k e r a t i t i s ,
w a s used t h r o u g h o u t t h e s t u d y ( 4 ) .
The i n f e c t i o u s
g e n e r a l l y h e l d , as a r e s u l t of b a s i c and c l i n i c a l
t i t e r s w e r e determined v i a plaque t i t r a t i o n .
o b s e r v a t i o n , t h a t c e l l u l a r immunity i s o f g r e a t
Anti-gD monoclonal a n t i b o d i e s
importance i n e r a d i c a t i n g h e r p e t i c i n f e c t i o n s .
We have a l r e a d y e s t a b l i s h e d 6 c l o n e s of anti-gD
However, a r e c e n t r e p o r t by Lausch and h i s
monoclonal a n t i b o d i e s ( 4 ) .
a s s o c i a t e s r e v e a l e d t h e s i g n i f i c a n t r o l e of
w e used 2 c l o n e s , d e s i g n a t e d M1 and M12, of t h e s e
antibodies i n e f f i c i e n t l y eliminating the v i r u s
antibodies.
I n t h e present study,
Received on September 13, 1991; accepted on December 9, 1991
@ Oxford University Press
53
Current
Eye Research E;val>i-&ion
o f e f f e c t s ( ~ i Fac;sive immunization w i t h
m n o-i l o n__ s i l___ antibodies - o-. Two
c
titers -~
l o ~ l e so t monoclonal a n t i b o d i e s (MI a n d
The n e u t r a l i z i n g a n t i b o d y t i t e r s were d e t e r m i n e d
M I L ) against g l y c o p r o t e i n D were p r e p a r e d as described pLeviously
((4).
by t h e p l a q u e r e d u c t i o n method i n a manner d e s c r i b e d p r e v i o u s l y ( 4j
UALB/r mice w e r e
.
i n t r a p e t i toned1 Ly i n o c u l a t e d w l t h d i f t e r e n t d o s e s
CDC a s s a y s
of monaLluna1 A n t i b o d i e s (1, 10, 25, 100 pg/mouse)
CDC a s s a y s were p e r f o r m e d as d e s c r i b e d p r e v i o u s l y
24 h o u r 9 b e € , j r r , Lmmediately, 24 h o u r s a f t e r ,
(4).
and rt8 h o i i r s a f t - e r v i r u s i n o c u l a t i o n .
HSV-infected
dnclsthe
bvst
emIi
:he
Ci,rrie3s
wit11
i
ranging
Downloaded by [New York University] at 17:48 07 April 2016
D e t e r m i n a t i o n of s e i - u m - n e u t r a l i z i n g a n t i b o d y
Following
wi t h s o d i u m p e n t o b a r b i t a l ,
> f t-lie m ~ c ew e r p s c a r i E i e d 20 t i m e s
? 6 g d u g e n e e d l e a n d i n f e c t e d w i t h HSV it1
titer,
f r o m 1 . 7 ~ 1 0t o~ 6 . 0 ~ 1 0p ~f u f e d ?yes w e r e examined w i t h
a liantl-slit
itimp i n masked f a s h i o n a s t o s e v e r i t y
