23p
Medical Research Society
75. FRACTIONATION OF FULMINANT HEPATIC
FAILURE SERUM TO
FURTHER CHARACTERIZE
THE
LEUCOCYTE
SODIUM
TRANSPORT
INHIBITOR
R. D. HUGHES, R. B. SEWELL, L. POSTON AND R. WILLIAMS
Liver Unit, King's College Hospital and Medical School,
Denmark HiI/, London S.E.5
The presence of toxic metabolites in blood is considered to be
responsible for many of the metabolic abnormalities in fulminant
hepatic failure. Previously it has been shown by our Unit that
serum from these patients is inhibitory to leucocyte ouabainsensitive sodium transport (Alam et al., Clinical Science and
Molecular Medicine, 55, 355). Inhibition of cellular ATPase
could be related to the production of cerebral oedema, which is
one of the major causes of death in fulminant hepatic failure.
The aim of this study was to attempt to further characterize the
serum inhibitors.
Membrane ultrafiltrates « 10 000 mol.wt.) of serum were
chromatographed on Sephadex G-25 with NH.HCO, (0·1
mol/I), with absorbance monitored at 206 nm and 254 nm.
Fractions were collected for toxicity assay on the efflux of
2lNa from pre-loaded normalleucocytes.
Similar toxicity to that in serum was observed in the
ultrafiltrate and the most toxic G·25 fraction (peak 2) was not
detected in pooled normal serum. Further chromatography of
peak 2 material on Sephadex G-15 gave three discrete peaks.
The second part of this study was to investigate the effect of
artificial liver support systems on the serum chromatographic
profiles. Material eluted from resin columns post-haemoperfusion and dialysis fluid post-polyacrylonitrile haemodialysis
both showed removal of peaks 2 and 4. This was further studied
in adsorption experiments in vitro with fulminant hepatic failure
serum.
With these techniques it is hoped to characterize the toxins
responsible for the defect in leucocyte sodium transport and
determine which method of artificial liver support will remove
them most efficiently.
76. MEAL-INDUCED ABSORPTION OF NEWLY FORMED SECONDARY BILE ACIDS IN NORMAL MAN
K. D. R. SETCHELL', A. M. LAWSON', E. J. BLACKSTOCK' AND
G. M. MURPHyt
'Mass Spectrometry Sub-division, Division of Clinical Chemistrv, Chemical Research Centre, Harrow, Middlesex HAl 3UJ,
and the tGastroenterology Unit, Guy's Hospital Medical
School, London SEl 9RT
Secondary bile acids are formed by the action of intestinal
bacteria on the conjugated primary bile acids. The first step in
the formation of these bacterial metabolites is usually deconjugation and many studies have shown that a large number of
unconjugated bile acid metabolites may be formed in normal
man. However, whilst there have been numerous studies of the
excretion of unconjugated bile acids, little is known of their
absorption. In the normal situation peripheral bile acid concentrations are determined largely by the input from the intestine
and therefore we have used serum unconjugated bile acid
concentrations as an index of the intestinal absorption of newly
formed secondary bile acids.
By using anion-exchange chromatography combined with gas
chromatography-mass spectrometry serum unconjugated bile
acid concentrations were measured in two subjects throughout
a 24 h period during which three meals were taken and in two
subjects before and during a 6 h period after breakfast. In the
latter studies glycocholic acid concentrations were also determined by using a radioimmunoassay.
Unconjugated bile acids were found in all samples and
included cholic, chenodeoxycholic, deoxycholic, allodeoxycholic, hyodeoxycholic, ursodeoxycholic, 3p-cholenoic and
lithocholic acids. Peak concentrations of all the metabolites
occurred after meals in all four subjects, levels remammg
relatively low during the overnight fast. The concentrations of
the individual unconjugated bile acids after breakfast were often
50% that of the conjugated bile acid, glycocholate, in the same
samples but in the absence of food returned to pre-breakfast
levels within 2 h whereas those of the conjugated bile acids
remained raised. These studies show for the first time that there
is a diurnal variation in serum unconjugated bile acid levels in
normal man and that the pattern of absorbed newly formed
secondary bile acids is complex and of quantitative importance
to normal serum total bile acid concentrations.
77. HLA-B8 AND CELLULAR IMMUNE RESPONSE TO
GLUTEN
F. G. SIMPSON,
LOSOWSKY
A. W. BULLEN, D. A. F. ROBERTSON AND M, S.
University Department of Medicine, Clinical Sciences Building,
St James's Hospital, Leeds LS9 7TF, U.K.
Ingestion of the wheat protein gluten causes jejunal mucosal
damage in coeliac disease. Most patients with coeliac disease
have the histocompatibility antigen HLA-B8. Measurement of
leucocyte migration inhibition has demonstrated increased
cell-mediated immunity to gluten in coeliac disease compared
with controls (Bullen & Losowsky, 1978, Gut, 19, 126). It has
been suggested that this is specific for coeliac disease and may
be helpful as a diagnostic test. However, lymphocytes from
healthy B8 individuals transform with gluten (CunninghamRundles et al., 1979. Transplantation Proceedings. 10, 977).
We have measured leucocyte migration index (LMI) to gluten
fraction III in 30 healthy controls (14 B8, 16 non-B8), 26
untreated coeliac patients (20 B8, six non-B8) and 57 treated
coeliac potentials (41 B8, 16 non-B8) and related the results to
HLA status and effects of a gluten-free diet.
In B8 controls there was significant depression of LMI
(mean O·88, SD 0·08) indicating increased immunity compared
with non-B8 controls (mean 0·98, SD 0·10, P < 0·02). Untreated coeliac patients, whether B8 or not, showed no difference
from B8 controls (mean 0·89, SD 0,13). Coeliac patients early in
treatment showed depression of LMI (mean O·78, SD 0·06,
P < 0·001), which improved after treatment for over I year
(mean 0·88, SD 0·08), suggesting that gluten-reactive lymphocytes are sequestered in the gut in untreated coeliac patients,
appear in the blood when gluten is excluded from the diet and
later decline in numbers. It is of importance that B8 and non-B8
coeliac patients behave similarly at all stages.
These results suggest that coeliac disease patients, whether B8
or not, are similar to B8 controls in their cellular immunity to
gluten and that this differs from that of non-B8 controls. This
pattern of response may represent an immune response gene for
cellular responses to gluten which is in linkage disequilibrium
with HLA-B8. Similar immune-response genes have been
described in animals but not in man. The pattern of immune
response to gluten described seems necessary but cannot be
sufficient for the development of coeliac disease. Previous results
with tbe LMI test in coeliac disease must be re-evaluated with
reference to the HLA status of the controls.
78. ENTEROPANCREATIC TROPHIC AXIS: A NEW
MODEL FOR STIMULATING ADAPTIVE PANCREATIC
HYPERPLASIA
H. LEVAN, B. M. MIAZZA, S. V AJA AND R. H. DOWLING
Gastroenterology Unit, Department ofMedicine, Guy's Hospital
and Medical School, London SEl 9RT
In order to further study the role of pancreatico-biliary
secretions (PBS) (Altmann, 1971, American Journal of
Anatomy, 132, 167-178) on the adaptive changes in structure
and function of the gut observed in many different experimental
situations, we diverted these PBS by transposing the duodenum