Sclerosing cholangitis andbiliary calculi -primary secondary?
Antineutrophil cytoplasmic antibodies after liver transplantation for primary sclerosing cholangitis...
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HEPATOLOGY Vol. 22, No. 4, Pt. 2, 1995 AASLD ABSTRACTS 391A 1137 CHANGES IN PROLIFERATIVE AND CYTOTOXIC ACTIVITY OF MNL ISOLATED FROM RAT LIVER IN STABLE G1 PHASE INDUCED BY THREE DAYS OF PROTEIN DEFICIENT DIET A Azzarone, L Polimeno. A Iacobellis, OH Zenm T Whiteside. T Starzl, and A Francavilla. Dept. of Surgeiy, University of Pittsburgh, Dept. of Gastroenterology, University of Bari. Recently we have demonstrated that twentyfour hours after 70% PH, the proliferation induced by Con A and IL-2, as well as the cytotoxie functions of liver resident mononuelear leukocytes (RMNL) are dramatically suppressed (J Immun 154:1-14, 1995). In this study we evaluated the immunological events occurring during phase G1 of the cell cycle. Stable G1 phase is induced in the rat by three days of protein deficiency diet (PDD) as reported by N. Bucher (In Cell Production, ICN/UCLA Symp on Mol Cell Biol, 112:661-670, 1978). MNL are isolated from the liver of rats treated with PDD for proliferative and cytotoxic activity studies. The proliferative activities of MNL are determined in vitro in the presence of differsnt doses of IL-2 and Con A and evaluated by incorporation of ~?I-Thymidine; the cytotoxic activities of MNL was evaluated by JICr release in a standard micro assay in which YAC-1 cells or hepatocytes were used as the target. The proliferatwe activity of MNL isolated from PDD rats is similar to that found using MNL from a control group in the presence of IL-2 but siglaiflcantly augmented in the presence of Con A (Con A 5ug/ml --aH-Thyrnidine incorporation (cpm/20,000 ceils normal liver 10915_+1891 - PDD liver 46523_.+6240). The cytotoxic activity of MNL in the PDD group was reduced by 55% in the presence of YAC-1 cells (365+64 LU/161 .+45 LU) and absent in the presence of hepatocytes isolated from rats fed with PDD. These results suggest that changes in the RMNL activity is an early event after PH and appears immediately after the beginning of the proliferation. This condition is not stable but changes during the other phases of the cell cycle as has been reported by us in S phase in rat liver 24 hours after PH (J Immun 154:1-14, 1995). These results show dynamic changes in the activity of RMNL in relation to the cell cycle of proliferating hepatocytes thus confirming the existence of mutual control between the two systems. 1138 PROSPECTIVE STUDY OF EXOGENOUS FACTORS IN INTRAHEPATIC CHOLESTASIS OF PREGNANCY. Y.Baco T Sapey. M-C Br6chot. F Pierre. A Fianon, and F Dubois. Centre Hospitalo-Universitaire de Tours, France. Although the cause of intrahepatic cholestasis of pregnancy (ICP), a rare disease in France (2-4 per 1000 deliveries), remains unknown, genetic factors and the role of estrogens are clearly associated with ICP. On the other hand, some authors have stated that exogenous factors may modulate the expression of the disease. The aim of this prospective study was therefore to identify such exogenous factors. From 1989 to 1995, we studied 45 consecutive pregnant women with ICP (22 primiparous, 4 twin and 2 triplet pregnancies, 9 recurrent ICP) referred for hepatologic consultation. All patients suffered from pruritus associated with elevated fasting serum levels of total bile acids (mean: 48 i.tmol/I, range: 7-290). No patients had concomitant liver disease and all recovered normal liver function after delivery. Serologic markers of viral hepatitis were negative. Hypertension, fever, and cervical and urinary infections were exclusion criteria. Prematurity rate was 51% and perinatal mortality was 4%. Systematic clinical interviews revealed that 32 patients had been treated with natural progesterone (200-1000 mg per day) during the current pregnancy for premature labour or cervical modifications, including 30 (67%) before the onset of pruritus. Twenty patients had also received salbutamol treatment. These results suggest that oral natural progesterone might be an exogenous factor which triggers ICP in predisposed women. Further investigations are necessary to confirm this hypothesis. 1139 ANTINEUTROPHIL CYTOPLASMIC ANTIBODIES AFTER LIVER TRANSPLANTATION FOR PRIMARY SCLEROSING CHOLANGITIS. D Bansil, B Gunson 2, J Neuberger 2, K Fleming3, R Chapman 1. Dept. of Gastroenterology 1 and Nuftield Dept. of Pathology 3, Oxford Radcliffe Hospital, Oxford UK. Liver Research Laboratories2 Queen Elizabeth Hospital, Birmingham, UK. The pathogenic role of antineutrophil cytoplasmic antibodies (ANCA) in primary sclerosing cholangitis (PSC) and Ulcerative colitis (UC) is unclear. We investigated the presence of ANCA before and after liver transplantatign for PSC. METItODS: 14 patients with end- stage PSC were studied (11 males, mean age 45 years, 8 with UC, 2 with Crohn's colitis). ANCA was detected by an immunoalkaline phosphatase technique, RESULTS: Perinuclear ANCA was detected in 7 (50%) patients prior to liver transplantation. Of these, 5 had persistent ANCA up to 18 months post-transplant. (In one, a rise in titre was noted from 1:5 to 1:200, while in two, the titre fell from 1:100 to 1:5.) In the remaining two, ANCA disappeared after transplantation. Seven patients were ANCA-negative prior to transplant and 6 of these remained negative for up to 2 years. In the remaining patient, ANCA became positive at 2 weeks. No recurrence of PSC was detected in any patient as judged by liver biopsy. CONCLUSION: ANCA is not significantly affected by liver transplantation, suggesting that synthesis is unrelated to the presence of the diseased organ. This argues against a pathogenic role for ANCA in PSC. 1140 DETECTION OF ANTINEUTROPHIL CYTOPLASMIC ANTIBODY IN PRIMARY SCLEROSING CHOLANGITIS : COMPARISON OF THE ALKALINE PHOSPHATASE AND IMMUNOFLUORESCENT TECHNIQUES. D Bansi 1, K Boberg2,U Broome 3, R Chapman 1, K Fleming 1, R Jorgnnsen4, K Lindor 4, E Schrumpf 2. John Radcliffe Hospital 1, Oxford, UK. The National Hospital 2, Oslo, Norway. Huddinge Univ. Hospital 3, Huddinge, Sweden. Mayo Clinic 4, Rochester, USA. The variation in prevalence of antineutrophil cytoplasmic antibodies (ANCA) in primary selcrosing cholangifis (PSC) may be methodological. To resolve this issue we compared the sensitivity (true pos.:true pos.+false neg. ratio) and specificity (true neg.:true neg.+false pos.ratio) of the indirect alkaline phosphatase (IALP) and immunofluorascence (IF) techniques. Method: Sera from 3 ¢entres were tested blinded on alcohol-fixed neutrophils using both techniques. Patients; USA:14 PSC,14 primary biliary cirrhosis (PBC); Sweden:32 PSC, 3 autoimmune hepatitis (AIH),14 PBC, I I chronic liver disease; Norway:32 PSC,14 AIH,13 PBC, 1 hop. C. 36 normals and positive and negative controls were included. Results: Sensitivity and specificity of ANCA for PSC (excl. controls) : Sensitivity 10114 (71%) 21/32 (66%) 22/32 (69%) Specificity 13114 (93%) 27128 (96%) [ 13/2g (46,%) Sweden 18)32(56%) I IF method USA ] Norway .Sensitivity 7/14 (50%) | 15132(47%) Specificity 12/14(86%) | 24/28(g6%) 17/2g (61%) Overall IALP sensitivity was 53/78 (68%) and specificity 53170 (76%), compared with 40/78 (51%) and 53/70 (76%) for IF respectively. Conclusions: Overall the IALP method of ANCA detection has greater sensitivity and equal specificity to IF for serological diagnosis of PSC. We recommend use of this method which is also easier to interpret. Future identification of the antigen(s) should allow development of a more sensitive and specific diagnostic assay for PSC.
Transcript of Antineutrophil cytoplasmic antibodies after liver transplantation for primary sclerosing cholangitis...
HEPATOLOGY Vol. 22, No. 4, Pt . 2, 1995 AASLD A B S T R A C T S 3 9 1 A
1137 CHANGES IN PROLIFERATIVE AND CYTOTOXIC ACTIVITY OF MNL ISOLATED FROM RAT LIVER IN STABLE G1 PHASE INDUCED BY THREE DAYS OF PROTEIN DEFICIENT DIET A Azzarone, L Polimeno. A Iacobellis, OH Zenm T Whiteside. T Starzl, and A Francavilla. Dept. of Surgeiy, University of Pittsburgh, Dept. of Gastroenterology, University of Bari.
Recently we have demonstrated that twentyfour hours after 70% PH, the proliferation induced by Con A and IL-2, as well as the cytotoxie functions of liver resident mononuelear leukocytes (RMNL) are dramatically suppressed (J Immun 154:1-14, 1995).
In this study we evaluated the immunological events occurring during phase G1 of the cell cycle. Stable G1 phase is induced in the rat by three days of protein deficiency diet (PDD) as reported by N. Bucher (In Cell Production, ICN/UCLA Symp on Mol Cell Biol, 112:661-670, 1978). MNL are isolated from the liver of rats treated with PDD for proliferative and cytotoxic activity studies. The proliferative activities of MNL are determined in vitro in the presence of differsnt doses of IL-2 and Con A and evaluated by incorporation of ~?I-Thymidine; the cytotoxic activities of MNL was evaluated by J ICr release in a standard micro assay in which YAC-1 cells or hepatocytes were used as the target.
The proliferatwe activity of MNL isolated from PDD rats is similar to that found using MNL from a control group in the presence of IL-2 but siglaiflcantly augmented in the presence of Con A (Con A 5ug/ml --aH-Thyrnidine incorporation (cpm/20,000 ceils normal liver 10915_+1891 - PDD liver 46523_.+6240). The cytotoxic activity of MNL in the PDD group was reduced by 55% in the presence of YAC-1 cells (365+64 LU/161 .+45 LU) and absent in the presence of hepatocytes isolated from rats fed with PDD. These results suggest that changes in the RMNL activity is an early event after PH and appears immediately after the beginning of the proliferation. This condition is not stable but changes during the other phases of the cell cycle as has been reported by us in S phase in rat liver 24 hours after PH (J Immun 154:1-14, 1995). These results show dynamic changes in the activity of RMNL in relation to the cell cycle of proliferating hepatocytes thus confirming the existence of mutual control between the two systems.
1138 PROSPECTIVE STUDY OF EXOGENOUS FACTORS IN INTRAHEPATIC CHOLESTASIS OF PREGNANCY. Y.Baco T Sapey. M-C Br6chot. F Pierre. A Fianon, and F Dubois. Centre Hospitalo-Universitaire de Tours, France.
Although the cause of intrahepatic cholestasis of pregnancy (ICP), a rare disease in France (2-4 per 1000 deliveries), remains unknown, genetic factors and the role of estrogens are clearly associated with ICP. On the other hand, some authors have stated that exogenous factors may modulate the expression of the disease. The aim of this prospective study was therefore to identify such exogenous factors. From 1989 to 1995, we studied 45 consecutive pregnant women with ICP (22 primiparous, 4 twin and 2 triplet pregnancies, 9 recurrent ICP) referred for hepatologic consultation. All patients suffered from pruritus associated with elevated fasting serum levels of total bile acids (mean: 48 i.tmol/I, range: 7-290). No patients had concomitant liver disease and all recovered normal liver function after delivery. Serologic markers of viral hepatitis were negative. Hypertension, fever, and cervical and urinary infections were exclusion criteria. Prematurity rate was 51% and perinatal mortality was 4%. Systematic clinical interviews revealed that 32 patients had been treated with natural progesterone (200-1000 mg per day) during the current pregnancy for premature labour or cervical modifications, including 30 (67%) before the onset of pruritus. Twenty patients had also received salbutamol treatment. These results suggest that oral natural progesterone might be an exogenous factor which triggers ICP in predisposed women. Further investigations are necessary to confirm this hypothesis.
1139 ANTINEUTROPHIL CYTOPLASMIC ANTIBODIES AFTER LIVER TRANSPLANTATION FOR PRIMARY SCLEROSING CHOLANGITIS. D Bansil, B Gunson 2, J Neuberger 2, K Fleming 3, R Chapman 1. Dept. of Gastroenterology 1 and Nuftield Dept. of Pathology 3, Oxford Radcliffe Hospital, Oxford UK. Liver Research Laboratories 2 Queen Elizabeth Hospital, Birmingham, UK.
The pathogenic role of antineutrophil cytoplasmic antibodies (ANCA) in primary sclerosing cholangitis (PSC) and Ulcerative colitis (UC) is unclear. We investigated the presence of ANCA before and after liver transplantatign for PSC. METItODS: 14 patients with end- stage PSC were studied (11 males, mean age 45 years, 8 with UC, 2 with Crohn's colitis). ANCA was detected by an immunoalkaline phosphatase technique, RESULTS: Perinuclear ANCA was detected in 7 (50%) patients prior to liver transplantation. Of these, 5 had persistent ANCA up to 18 months post-transplant. (In one, a rise in titre was noted from 1:5 to 1:200, while in two, the titre fell from 1:100 to 1:5.) In the remaining two, ANCA disappeared after transplantation. Seven patients were ANCA-negative prior to transplant and 6 of these remained negative for up to 2 years. In the remaining patient, ANCA became positive at 2 weeks. No recurrence of PSC was detected in any patient as judged by liver biopsy. CONCLUSION: ANCA is not significantly affected by liver transplantation, suggesting that synthesis is unrelated to the presence of the diseased organ. This argues against a pathogenic role for ANCA in PSC.
1140 DETECTION OF ANTINEUTROPHIL CYTOPLASMIC ANTIBODY IN PRIMARY SCLEROSING CHOLANGITIS : COMPARISON OF THE A L K A L I N E P H O S P H A T A S E AND IMMUNOFLUORESCENT TECHNIQUES. D Bansi 1, K Boberg2,U Broome 3, R Chapman 1, K Fleming 1, R Jorgnnsen 4, K Lindor 4, E Schrumpf 2. John Radcliffe Hospital 1, Oxford, UK. The National Hospital 2, Oslo, Norway. Huddinge Univ. Hospital 3, Huddinge, Sweden. Mayo Clinic 4, Rochester, USA.
The variation in prevalence of antineutrophil cytoplasmic antibodies (ANCA) in primary selcrosing cholangifis (PSC) may be methodological. To resolve this issue we compared the sensitivity (true pos.:true pos.+false neg. ratio) and specificity (true neg.:true neg.+false pos.ratio) of the indirect alkaline phosphatase (IALP) and immunofluorascence (IF) techniques. Method: Sera from 3 ¢entres were tested blinded on alcohol-fixed neutrophils using both techniques. Patients; USA:14 PSC,14 primary biliary cirrhosis (PBC); Sweden:32 PSC, 3 autoimmune hepatitis (AIH),14 PBC, I I chronic liver disease; Norway:32 PSC,14 AIH,13 PBC, 1 hop. C. 36 normals and positive and negative controls were included. Results: Sensitivity and specificity of ANCA for PSC (excl. controls) :
Sensitivity 10114 (71%) 21/32 (66%) 22/32 (69%) Specificity 13114 (93%) 27128 (96%) [ 13/2g (46,%)
Sweden 18)32(56%) I IF method USA ] Norway .Sensitivity 7/14 (50%) | 15132(47%) Specificity 12/14(86%) | 24/28(g6%) 17/2g (61%) Overall IALP sensitivity was 53/78 (68%) and specificity 53170 (76%), compared with 40/78 (51%) and 53/70 (76%) for IF respectively. Conclusions: Overall the IALP method of ANCA detection has greater sensitivity and equal specificity to IF for serological diagnosis of PSC. We recommend use of this method which is also easier to interpret. Future identification of the antigen(s) should allow development of a more sensitive and specific diagnostic assay for PSC.
