LIVER TRANSPLANTATION BEFORE ONE YEAR.
E Sokal, J d e Ville de Goyet, D Moulin,
F Veyckemans,L Van Obbergh, M Carlier,
D Latinne, J P Buts, J Rahier, J B Otte.
University of Louvain St Luc Hospital, Brussels.
Young
infants
are
often
refused
for
liver
transplantation on the sole basis of young age. Over
a total of 141 chidren who received an orthotopic
liver transplantation in our centre (March 1984-July
1989), 17 patients (I?%!, were transplanted before
they 1st birthday (15 blllary atresia, 1 Byler disease
& 1 tyrosinemia)
Mean age was 10,3 months (range 811) and mean weight 7,3 kg (range 5,2 - 13). A reduced
liver was used
11 times over a total of 26
transplantations
(42%). Immunosuppression
included
cyclosporine A, prednisone and azathioprine & OKT3 or
ATG for sterold resistant rejection. Survivors were
discharged after a mean hospital stay of 47 days
(range 22-87) and the one year actuarial survival is
65% versus 77% in the whole serie. Reasons for
retransplantation (29%) were primary non function ( 3 ) ,
hepatic artery thrombosis (4) and rejection (2). 1
patient died perioperatively, 2 from primary non
GENETIC STUDY OF HUMAN ADULT-TYPE HYPOLACTASIA BY ANALYSIS OF
RESTHlCTlONFRACMENTLENCHTPOLYMORPHISMSlRFLPa)OFTI1ELACTASE
17
.
function,
1
from
adenovirus
infection,
from
2
rejection & 1 from bone marrow aplasia. 18 rejection
episodes,
(11
corticoresistant)
occured
in
11
patients. Liver transplantation can be proposed t o
young infants without age limits being imposed.
Scmenra* and S . A u r i c c h i o .
Naplrs,ltaly;
*Dept.
Dept. o f P e d i a t r i c s . 2 n d
o f Biochemistry,
GENE.
Vizia,A.Ballabio.Vl.Doll*,N.Mantei*,G.
L.Sebartio,V.Cuzzetta,8.0e
School o f M e d i e i n e , U n i v e r s i t y
Swiss F s d e r a l
of
I n s t i t u t e o f Technology, ETH-
Zentrum,Zurich,Switzerland.
The a d c l t - t y p e h y p o l a c t a s i a i s a c o n d i t i o n due t o h o r n o z i g o s i t y f o r a r e c e s s i v e
a l l e l e o f a h y p o t h e t i c r e g u l a t o r y gene w h i c h c o n t r o l s t h e l a c t a s e a c t i v i t y i n t h e
adulthood.
I t i s s t i l l u n c l e a r whether t h i s r e g u l a t o r y gene i s p a r t a f t h e l a c t a s e
gene i t s e l f o r maps t o a d i f f e r e n t
locus.
To i n v e s t i g a t e t h i s aspect,
we have used
a g e n e t i c approach: t h e a n a l y s i s o f f a m i l y s e g r e g a t i o n o f b o t h t h e l a c t a s e gene
and t h e l a c t o s e a b s o r p t i o n c a p a c i t y , w h i c h c o r r e l a t e s w i t h t h e l a c t a s e a c t i v i t y .
8 I t a l i a n f a m i l i e s ( 2 0 meioses) e n t e r e d t h i s s t u d y . F a m i l ~ r , were chosen on t h e
b a s i s o f t h e c o e x i s t e n c e o f s u b j e c t s w i t h e i t h e r h y p o l a c t a s i a or p e r s i s t e n c e o f
h i g h lactaae a c t i v i t y .
The l a c t o s e a b s o r p t i o n c a p a c i t y was assessed by b r e a t h - t e s t
a f t e r an o r a l l o a d o f 50 g o f l a c t o s e .
To f o l l o w t h e l a c t a s e gene s e g r e g a t i o n ,
we
searched f o r RFLPs i d e n t i f i e d b y t h e l a c t a b e cDNA. DNA o f each s u b j e c t was
d i g e s t e d by t h e f o l l o w i n g r e s t r i c t i o n enzymes, w h i c h we found t o d e t e c t RFLPr o f
t h e l a c t a s e gene: P s t I, Msp I, Rsa I, B c l I. i n a l l t h e i n f o r m a t i v e p e d i g r e e s ,
c o s e g r e g a t i o n o f t h e l a c t a r e gene and t h e h y p o l a c t a s i c c o n d i t i o n has beenobserved.
T h i s r e s u l t suggests t h a t an e v e n t i n v o l v i n g t h e l e c t a s e gene i t s e l f (e.9.
r e g u l a t i o n by sequences a t t h e promoter;
l e a d t o e i t h e r h y p o l a c t a s i n or,
activity.
condition,
d i f f e r e n t RNA e d i t i n g o r s p l i c i n g ) can
by m u t a t i o n ,
t o the persisLrnce of high I ~ l c t a s e
Indeed, we cannot e x c l u d e t h a t h y p o l a c t a s i a m i g h t be e heterogeneous
i n w h i c h a l s o o t h e r mechanisms (e.g.
post-translational modification)
may c o n t r o l i n t r a n s - t h e l a c t a s e gene e x p r e s s i o n .
MOSAIC PAlTERN OF LACTHSE E7PRESSION BY VILLUS LNTEROCYTES IN HUMAN
ADULT-TYPE HYPOLACTASIA. L . n a i u r i r
V.Raia*
J.Potter"
D.Snallow;
MWan H o " . R . F i o c c a " ~ " . C . F i i z i i " , M n ~ ~ r n a q q i a ~ ~ ~ ~ ~ . C . C a 0 ~ 1 1 a ~ " ,
A.Ouaroni"^^.S.Auricchio*.
of Naples,Naples,Italy;
*Department
of Pediatrics,
OBiology D i s c i p l i n e , O p e n
1 1 Medical Schaol,University
U n i v e r s i t y , M i l t o n Keynes,
England; ^MRC,Human B i o c h e m i c a l G e n e t i c s U n i t , U n i v e r s i t y C o l l e g e o f London,
Londan,England;
..
V a r e s e.. l t a l v :
"IRCCS P a l i c l i n i c o 5. H a t t e o , P a v i a , l t a l y ;
""Multironal
Hospital,
.
""Deoartment
o f Human P a t h o l o a-,
v .. U n i v e r s i t v o f Pavia.Pavia. . I t a l v,,
:
.
"^^Section o f Physiology. C o r n e l l U n i v e r s i t y ,
l t h a c a , N.Y.,
CHLORIDE SECRETION PAlTERNS IN RECTAL EPI'IliELIUM OF
CYSTIC FIBRCCjIS (CF) PATIAND ITS RELATION TO 'R1E
H.
h J.
F508
Veeze
tWl?p"l'ON.
, M Sinaasappelx, D.J. J. HaUeyl,
J. KIUW@C', J. tllll~n-',ti. K ae donqe-.
'dept. Pediatrics sutd. Gastroenteml
E%?.smus University
ard
'dept.University
Clinical~osbital
Genetics,Rotterdam
Di ' +ig( So
A phia
i t awld r Rotterdam
e n ' s Hcspital.
3dept Cellbid
E ~ ~ ~ I Ihruversity
IUS
Rotkerdam
4dept: ~iochemis*r&, Eraemus Univ., ~okterdam,The Netherlands.
21
USA.
E i g h t d i f f e r e n t monoclonal a n t i b o d i e s w h i c h r e c o g n i z e a t l e a s t t h r e e d i s t i n c t
e p i t o p e s u n i q u e t o t h e l a c t a r e p r o t e i n o f human e n t e r o c y t e have besn used t o
i n v e s t i g a t e l a c t a r e p r o t e i n e x p r e s s i o n i n h y p o l a c t a s i a o f N e a p o l i t a n a d u l t s by
l i g h t and e l e c t r o n m i c r o s c o p y w i t h t h e immunogald t e c h n i q u e .
A l l t h e a n t i b o d i e s gave t h e same r e s u l t s ,
namely: s t r o n g b r u s h b o r d e r s t a i n i n g i n
a l l t h e 7 l a c t a r s p e r s i s t e n t a d u l t s t e s t e d ; no s t a i n i n g a t a l l i n 9 o f t h e
h y p o l a c t a s i a s u b j e c t s and a mosaic p a t t e r n o f s t a i n i n g o f e n t e r o c y t e s i n I
12 a d u l t s w i t h h y p o l a c t a r i a .
- ,ther
The p e r c e n t a g e o f e n t e r o c y t e s showing i n t e n s e
s t a i n i n g f o r l a c t a r e p r o t e i n v a r i e d between 4 and 20% o f t h e t o t a l v i l l u s c e l l s
(mean value+SO:9.86+5.84).
sampler examined.
l n t r a c e l l u l a r s t a i n i n g w a r n o t a p p a r e n t i n any of
the
S u c r a r e - i s o m a l t a s e p r o t e i n i n c o n t r a s t showed no mosaicism.
The mosaic p a t t r r n o f e x p r e s s i o n o f l a c t a s e p r o t e i n i n same h y p o l a c t a s i c s u b j e c t s
suggests t h a t columnar c e l l s d i s p l a y a no" homogeneous d i f f e r e n t i a t i o n a l o n g t h e
villus.
I f t h e two p a t t e r n s , m o s a i c i s m o r absence o f d e t e c t a b l e l a c t a s e p r o t e i n ,
are present along the e n t i r e small i n t e s t i n e o f the i n d i v i d u d i s tested, t h e
r e s u l t s w o u l d suggest t h a t two phenotypes o f a d u l t l h y p o l a c l a s i a e x i s t i n t h e
p o p u l a t i o n we s t u d i e d .
19
MOSAICISM OF BLOOD GROUP SPECIFICITY OF BRUSH BORDER OF HUMAN
ENTEROCYTES. S.Auricchio*.L.Maiuri*,V.Raia*.H.Fi0~~a~~".E.So1cia~,
G.Finzi""^.M.Cornaq4iat"'~
D.Swa1low-:
CONCLUSIONS:
Cb' recta^ epithelium of hmzygotes for ,F508 showed an atsent
Ca-mediated chloride secretion corresponding mth earlier studies
of small intestine and proximal colon One third of the CF
patients however showed residual chloride secretion; these
patients were hetemygotes,for AF508: In 2 compound heteroz gotes without residual chlor~de secretion we suggest that d e
other mutation is of the same severity as ~F508.
*Department o f P e d i a t r i c s , l l
O.NorCn'.H.SiijrtrBm'.H.Skavbiergl,
M e d i c a l School, U n i v e r s i t y o f Naples,
Naplen.ltaly;
"IRCCS P o l i c l i n i c o 5. M a t t e o , P a v i a , l t a l y ;
Varese.ltaly;
"Department of Human P a t h o l a g y , U n i v e r s i t y o f P a v i a . l t a l y ;
ment o f B i o c h e m i s t r y C, Panum I n r t i t u t e , U n i v e r s i t y
""Elulti2onal
Hospital,
'Depart-
of
the enterocyte along the v i l l u s i s considered
t o be dependent on t h e age o f t h e c e l l and u n i f o r m i n c e l l s o c c u p y i n g comparable
l o c a t i o n an t h e v i l l u r . Using a p o l y c l a n a l antibody against A blood group
s p e c i f i c components o f e n t e r o c y t e b r u s h b o r d e r and t h r e e monoclonal a n t i b o d i e s we
have demonstrated t h e e x p r e s s i o n o f t h e A and B a n t i g e n s i n t h e b r u s h b o r d e r and
Golgi apparatus of enterocytes of
l i g h t and e l e c t r o n micrascopy,
i n d i v i d u a l s o f t h e e x p e c t e d b l o o d groups, by
with
Dept. of Immunology, h~stituteof Child Ilealth, London WC IN. U.K.
o f Copenhagen, Denmark;^^MRC
Human B i o c h e m i c a l G e n e t i c s U n i t , U n i v e r r i t y C o l l e g e London, London, U.K.
The p a t t e r n o f d i f f e r e n t i a t i o n
22
THE GENERATION OF A TOLEROGEN A V E R TIIE
INGESTION OF OVALUUMlN IS TIME DEPENDENT AND
UNRELATED TO SERUM LEVELS OF IMMUNOREACI'ICE
ANTlGEN
Stephan Strobel, 110-Jen Peng, M a l ~ ' o l mW . Turner.
t h e immunagold t e c h n i q u e . The p o l y c l o n a l
a n t i - A serum was s p e c i f i c f a r t h e b r u s h b o r d e r whereas The monoclonal r e a g e n t s
a l s o bound t h e e n d o t h e l i a l c e l l s and e r y t r o c y t e r i n t h e s e c t i o n s . Four o f 16
b l o o d g r o u p A i n d i v i d u a l s showed a mosaic p a t t e r n o f e x p r e s s i o n o f t h e A a n t i g e n ,
b a t h w i t h t h e p o l y c l o n a l serum and t h e monoclonal a n t i - A r e a g e n t .
Same e n t e r o c y t e s showed a normal s t a i n i n g and some o t h e r e n t e r o c y r e s a t s i m i l a r
p o s i t i o n on t h e v i l l u s showed no s t a i n i n g a t a l l . An a n t i - L e a a n t i b o d y showed
t h a t a l l t h e 4 i n d i v i d u a l s w i t h t h e mosaic p a t t e r n were rm s e c r e t o r . A b n a r m a l i t i e s
o f b l o o d g r o u p a n t i g e n s may r e f l e c t abnormal e x p r e s s i o n o f g l u c o s y l - t r a n s f e r a s e
genes o r d i s t u r b a n c e s o f t h e i r a c c e p t o r s .
E n t e r o c y t e s may t h e r e f o r e d i s p l a y a non homogeneous p a t t e r n o f d i f f e r e n t i a t i o n
i n m o r p h a l o g i c a l l y s i m i l a r c e l l s o c c u p y i n g comparable l o c a t i o n an t h e v i l l u s .
I>uring our ~c.se;~rtl~
~ntothe regulauoi~ol the gastroinlebt~~:aI
~ I I I I I I I I I I\ybte811
~
(GA1.T) we have b~vcs~icatctl
lhu nic~leculdrad h r ~ ~ l o ~t ~. ~
udl r u r otovalbunis~
rs
(OVA) which has been 6bjecte<lto iriresri,lall~roce.ss~ig.
I~n~i~unoreactive
OVA absorbed by the gut was measured by a sandwich ELISA
at different times ,after feeding 25mg OVA to adult mice. OVA was detected ns
early as 5 n~inulesafter the feed (36.7 + I hnghnl, nlean + I SD) and reached
nlnrimal levels at I hour (73.3 + 20ngjnil). Pooled mouse serum, collected 5
n~inutesor I hour after the feed of OVA, was transferretl i.p. into naive
recipients. Suppression of syste~nicdelayed hypersensitivity (DTlI) was found
in inice receiving O.8n1l of serum obtained I hour after OVA feetling but not
when using serum obtained 5 minutes alter feeding. In order to triulsfer serunl
samples containing similar levels of OVA, an increased anount (1.31111)of seru~ii
collected 5 ~nirlutespost OVA feed was used in further experiments but ag:~in
failed to induce DTll tolerance. Serum svnples obtained 5 and 60 tninutes after
OVA feetling were a~alysedby fast protein liquid chro~natogr;~phy
(FI'LC)
;mJ tnolecul;~
fractiotlntion followed by ELISA. BOIIIthe charge cl~arac~eristics
weight of intestinally nhsorbed OVA were i~~Jisti~~guislial~lc
from native OVA.
' f i e results suggest tl~iu,althoufi11intact native OVA is the only n~olecularspecies
detected by ELISA, it h;u; no role in the suppression of DI'H responses, and Ih;ir
the gut-processed tolerogen may be modified so that less than two antibodybinding epitopes remain.
.