Al178 AASLD ABSTRACTS GASTROENTEROLOGY, Vol. 108, No. 4

Shedding New Light on Cholesterolosis and Cholelithiasis. W.P. Stuppy. Los Angeles, CA

First described grossly by Virchow, clinically b~ McCarty, and histochemically by Boyd, the relationship between the mucosal lipoid found in "strawberry gallbladder" and intraluminal cholesterol cholelithiasis remains controversial. The two substances were compared under routine and alternative forms of illumination; brilliant (BLM), polarized (PLM), ultraviolet (UVM) and fluorescent light microscopy (FLM).

Frozen Sections (2 micron) of fresh and formalin fixed specimens of gallbladder mucosa from eight patients with cholecystitis and cholesterolosis, three of whom also had cholesterol calculi, were compared-with a finely ground preparation made from the stones under BLM, PLM, UVM, and FLM with filtration for fluorescein isothiocyanate and tetramethylrhodamine isothio- cyanate.

All preparations exhibited classic findings identical to Boyd's under BLM and PLM. They also exhibited luminescence under UVM and auto- fluorescence under FLM.

These unique optical properties shared by gallbladder lipoid and stone material snrongly suggests chemical homogeneity and underscores the idea that they share a common pathogenesis. The propensity to develop one and/or the other may be predicated by lyotropic influences. Employing alternative forms of illumination might allow further retrospective correlation of the physical state, chemical composition, and genesis of cholesterolosis and cholesterol cholelithiasis.

GAMMA INTERFERON DOWN REGULATES RAT HEPATIC LIPOPOLYSACCHARIDE BINDING PROTEIN EXPRESSION. G.L.

Q. Wang, D. Williams, S.C. Wang. Departments of Medicine and Surgery, University of Pittsburgh, Pittsburgh PA.

Lipopolysaceharide~(LPS ) binding protein (LBP), plays an integral role in host responses to LPS~ " Little is known about the factors which regulate hepatic LBP pr0duction other than its induction in the acute phase response. la particular, little is known about the factors which down regulate LBP. In this study we examined the effect of IL-1, 1I.-6, tumor necrosis factor (TNF)~ 11.,-10, and Interferon-~t (IFN-'f) on LBP steady state mRNA level in primary cultures of isolated rat hepatocytes. Rat bepatocytes were isolated by in s tu c~llagenase perfusion and plated overnight. 'The hepatocytes were subsequently stimulated with varying concentrations of IL,1, IL-10, IL-6, TNF, and iFN-), in the presence of 10-7M dexamethasone for 24 hours prior to isolation of total RNA. Northern blot analysis was performed using 32p labeled eDNA probes for rat LBP, acid glycoprotein (AGP) and 18S. Quantitation of message level was determined using phosphorous imager. Variations in sample loading were corrected by expressing value as a ratio of LBP or AGP mRNA levels over 18S levels for each individual sample. IL-1 increased the steady state LBP mRNA levels by 2.5 fold. IFN-'¢ decreased steady state LBP mRNA levels in a dose dependent fashion. When the Northern blots were analyzed for AGP, another acute phase protein, similar decreases were also noted.

IFN-y IFN-'~ IFN-y IFN-~/ Control l0 U/ml 30 U/ml 100 U/ml 300 U/ml (n=4) (n=4) (n=4) (n=4) (n=4)

LBP/ 1.004#- .843#- .802#- .650+ .675+ 18S .005 .077 .043 .094 .087

*p~.0l * p~.0l * p<_.01 AGP/ 1.001+ .749+ .791#. .5384- .6684- 18S .0002 .044 .155 ,019 .t 17

*p~_.155 * p<_.O1 *p = compared to control.

Viability of cells was examined using a MTT assay which showed no difference between control and treated groups, therefore the observed decreases ~in LBP and AGP mRNA levels were not due to cell death. Investigative work has precisely focused on the factors which stimulate expression~of the acute phase proteins. However, very little is known about the fact~m which down regulate this response. In this study, we demonstrated for the first time that recombinant rat IFN-~, decreases steady state mRNA levels for LBP although the expresston is not completely abolished. A similar response is noted in AGP mRNA levels. These findings suggest a very important role for IFN-y in the down regulation of acute phase responses in the liver.

THE F R E Q U E N T O C C U R R E N C E OF A N T I - T H Y R O I D M I C R O S O M A L A N T I B O D I E S IN A U T O I M M U N E H E P A T I T I S . G.L.Su. R.Hall, K.Abu-Elmagd, J.Fung, J.Rakela. Departments of Medicine and Surgery, University of Pittsburgh, Pittsburgh, PA.

Autoimmune hepatitis is characterized by unresolving periportal hepatitis and the presence of autoantibodies. The autoantibodies most commonly reported in the United States, antinuclear antibodies (ANA) and anti- smooth muscle antibodies (SMA), are neither pathogenic nor disease specific. In this study, we examined the presence of these antibodies and those against the thyroid, anti-thyroid microsomal antibody (ATM) and anti-thyroglobulin antibody (THY), given the high incidence of thyroid disease in aul~immune hepatitis. 371 patient s were identified through the University of Pittsburgh Medical Archival System who had liver biopsy reports where the diagnosis of autoimmune hepatitis was considered. Of these patients, 77 patients were identified who'were given the diagnosis of autoimmune hepa[itis by their physician and had a score of > 10 by the criteria established by the International Autoimmune Hepatitis Group suggesting probable autoimmune hepatitis. 30 patients of the 77 had a score of > 15 which placed them as definite autoimmune hepatitis. The occurrence of autoantibodies in our population is as follows:

Score ANA SMA ATM THY > 10 53% 44% . 55% 2 2 % ( n = 7 7 ) (38/72) (31/70) (38/69) (13/60) >15 68% 60% 54% 18% (n = 30) ; . '' (19/28) (18/30) (15/28) (4/22)

The percentage of patients with elevated anti-thyroid microsomal antibodies is greater than or similar to those who are positive for ANA and SMA in our population. This is surprising since the study is biased toward finding a higher frequency of ANA and SMA given that elevations of these antibodies increase the score significantly in the criteria as established by the International Autoimmune Hepatitis Group. Of~,en more importance is the finding that in the group of patients who had all three antibodies evaluated, ANA, SMA, and ATM, 16% (10/63) had only an isolated anti- thyroid microsomal antibody (ATM). The significance of the high frequency of anti-thyroid antibodies in our population is as yet unknown but suggests an importance in the evaluation patients with autolmmune hepatitis that was previously not appreciated.

• HEPATITIS C VIRAL RNA IN BODY FLUIDS AFTER LIVER TRANSPLANTATION. M, $pe, S.H. Caldwell, J.H. Bowden, R.C. Dickson, C. DriscoU, W.C. Stevenson, M.B. Ishitani, C.S. MeCunough, T.L. Pruett, M. Lovell. Dept.s. Medicine, Pathology and Surgery. University of Virginia Medical Center, Charlottesville, Virginia.

Hepatitis C virus (I-ICV} has been infrequently detected in body fluids of patients (pts) with chronic infection. High serum levels of HCV have been observed iil pts with recurrent HCV after orthotopie liver transplantation (OLT). With the resumption of sexual activity after OLT, we have been concerned that the risk of sexual transmission may be increased. PURPOSE: To detect the presence of virus in body fluids, we measured HCV RNA in body fluids of 21 post-OLT pts (16 male, 5 female) on 2 or 3 drug immunosuppression and 8 HCV-infected. non-transplanted controls (3 male. 5 female). METHODS: We studied serum and 135 fluid specimens by the reverse transcriptase polymerase chain reaction (PCR) using nested primers for the 5' non-coding region. Fluid samples were collected and frozen at -70* until assayed. There were 29 sputum, 29 saliva, 8 vaginal, 29 tear, 29 urine and 11 semen samples. RNA was isolated from 200 ul aliquotes with a Quiamp column (Qiagen, Inc. Chatsworth, Ca) and eluates were concentrated by speed-vac centrifugation prior to PCR. Serum HCV-RNA was quantitated in eq/ml X 105 by the branched chain DNA assay (bDNA, Chiron). RESULTS: All pts and controls were serum PCR positive. bDNA in the post-OLT pts was 255 4- 229 compared to 71 + 57 in the controls (p= .002). Among the controls, all body fluids (8 sputum, 8 saliva, 3 vaginal, 8 tear, and 8 urine samplesl were negative for HCV RNA except one weakly positive vaginal specimen. In the post-OLT pts, 5 of 21 (24%) had positive body fluid RNA; 5 of 21 (24%) sputum, 4 of 21 (19%) saliva and 1 of 5 (20%) vaginal specimens were positive. All others were negative, bDNA in the 5 post- OLT pts with positive fluids was 460 + 157 compared to 191 4- 212 for the. 16 post-OLT pts with negative fluid samples (9=.014). CONCLUSIONS: HCV is more often detectable in saliva or sputum ofpts with recurrent HCV following OLT compared t~ non-OLT HCV-infected controls. Those with positive HCV- RNA in 10edy fluids have higher levels of viremia than those without detectable viral RNA in body fluids. It is not known whether these patients are at increased risk of transmitting the virus.