Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Reexamination Certificate
2000-03-28
2004-09-07
McGarry, Sean (Department: 1635)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
C435S006120, C536S023100, C536S024300
Reexamination Certificate
active
06787303
ABSTRACT:
1. INTRODUCTION
The present invention relates first, to the identification of a retrovirus and the novel nucleotide sequences encoding a retroviral polymerase gene (POL nucleotides) associated with the existence of primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH) Crohn's disease and ulcerative colitis. The present invention further relates to methods for using the PSC associated retroviral nucleotides for the detection of PSC, AIH, Crohn's disease and ulcerative colitis in patient samples. The present invention also relates to methods for using and targeting the PSC associated retroviral POL nucleotides in gene therapy protocols for the treatment of PSC, AIH, Crohn's disease or ulcerative disease in patients in need of such treatment. The present invention further relates to diagnostic protocols and kits for the detection of PSC, AIH, Crohn's disease and ulcerative colitis in tissue samples.
2. BACKGROUND OF THE INVENTION
Extraintestinal complications of inflammatory bowel disease, including diseases of the liver and biliary tract, affect a large number of patients with ulcerative colitis Crohn's disease. Indeed, hepatobiliary complications may affect as many as 10 percent of patients with inflammatory bowel disease. The occurrences of specific hepatobiliary diseases associated with inflammatory bowel disease, however, vary widely; some disorders occur commonly and others rarely. The clinical significances of individual hepatobiliary disorders also vary, ranging from inconsequential to life-threatening. Current data indicate that primary sclerosing cholangitis (PSC) is the most common disease associated with either ulcerative colitis or Crohn's disease. (Harmatz A., Hepatobiliary Manifestations of Inflammatory Bowel Disease. Med. Clin. North AM 78:1387, 1994).
Primary sclerosing cholangitis (PSC) was originally defined as a chronic cholestatic liver disease characterized by fibrosing inflammation of segments of the extrahepatic bile ducts. PSC results in a progressive narrowing or obliteration of bile duct lumens, progression to secondary biliary cirrhosis, with complications of portal hypertension, hepatic failure and cholangiocarcinoma is an idiopathic disorder characterized by inflammation and obliteration of both intra-hepatic and extrahepatic bile ducts. Cholangiocytes account for 3%-5% of the hepatic cell population and line a complex network of interconnecting conduits in the liver, termed the intrahepatic biliary ductal system. One of the pathological conditions manifested in both the intrahepatic and extrahepatic bile ducts is PSC.
The natural history of PSC is incompletely understood and the variability in the location of sclerotic lesions results in a wide spectrum of clinical disease. This is especially true for asymptomatic patients and the subgroup of patients with disease confined to the intrahepatic ducts. Although as many as 30 percent of patients with the disease have demonstrable autoantibodies, there is no serologic marker associated with PSC and the diagnosis is made by radiographic demonstration of irregular strictures and dialations within the bile ducts (Vierling, Hepatobiliary complications of ulcerative colitis and Crohn's disease. Third Edition. Philadelphia: W. B. Saunders, 1996). Patients invariably develop cirrhosis, the sequelae of liver failure, and 5 to 15 percent succumb to cholangiocarcinoma. At present, there is no curative medical therapy for PSC and liver transplantation is required to avoid these sequelae of endstage liver disease (Vierling, Hepatobiliary complications of ulcerative colitis and Crohn's disease. Third Edition. Philadelphia: W. B. Saunders, 1996).
In North America approximately 1 in 20,000 suffer from PSC and the majority of these patients have macroscopic or microscopic evidence of either ulcerative colitis or Crohn's disease (Vierling, Hepatobiliary complications of ulcerative colitis and Crohn's disease. Third Edition. Philadelphia: W. B. Saunders, 1996). Other hepatic disorders may occur in patients with inflammatory bowel disease in the absence of cholangiographic evidence of large bile duct disease, including autoimmune hepatitis (AIH) and pericholangitis, a non-specific histologic diagnosis of small duct PSC. (Vierling. Hepatobiliary complications of ulcerative colitis and Crohn's disease. Third Edition. Philadelphia: W. B. Saunders, 1996). AIH, pericholangitis, and PSC form the spectrum of the same hepatobiliary disorder associated with inflammatory bowel disease that may share a common etiological agent.
Autoimmune Hepatitis
An additional disease to which the present invention is directed is autoimmune hepatitis (AIH). Because there is no single diagnostic test for autoimmune hepatitis, it may initially be indistinguishable from other liver disorders. Early recognition is important, however, since patients suffering from autoimmune hepatitis benefit from treatment with immunosuppressives but not from treatment with interferons, used in the treatment of viral-induced hepatitis. Thus, the differentiation between viral-induced hepatitis and autoimmune hepatitis is important to ensure correct treatment. Autoimmune hepatitis, in particular in children, is an inflammatory disease which progresses into cirrhosis and hepatic insufficiency, which generally responds to an immunosuppressive treatment and is characterized by the presence of high non-organ-specific autoantibody titres.
Two subgroups have been defined, as a function of the autoantibody present in the serum: anti-smooth muscle antibody (anti-SMA) and anti-liver-kidney-microsome antibodies (anti-LKM), called hereinafter anti-SMA and anti-LKM antibodies.
The antigen recognized by the anti-LKM antibodies is a protein with a molecular weight of 50 kDa, which is present at a relatively high concentration in the endoplasmic reticulum. Several studies suggest that this antigen corresponds to a protein of the cytochrome P450 family (Waxman, 1988, Gastroenterol., 95: 1326). This was confirmed by screening a rat liver cDNA library in the presence of an anti-50 kDa protein antibody, purified by affinity from an LKM-positive serum. Both constitutive forms of cytochrome P450, belonging to the subfamily IID 1 and 2 (rat db1 and db2), have been identified, as the antigens recognized by the anti-LKM antibody (Gueguen et al., 1988 Exp. Med., 168: 801).
Other studies (Gueguen et al., 1989, Biochem. Biophys. Res. Commun, 159: 542; Zanger et al., 1988., Proc. Natl. Acad. USA, 1988, 27: 8256; Manns et al., 1989 J. Clin. Invest, 83:, 1066) have shown that cytochrome P450 IID6 is a protein recognized in human liver by the anti-LKM antibody. (Gueguen et al., 1989, Biochem. Biophys. Res. Commun, 159: 542; Zanger et al., 1988., Proc. Nati. Acad. USA, 1988, 27: 8256; Manns et al., 1989 J. Clin. Invest., 83:, 1066). Recently a solid-phase immnunoassay for the detection and quantification of LKM autoantibodies has been described (U.S. Pat. No. 5,741,654, Michel et al.) Other tests have therefore been proposed for the detection of auto-antibodies, in particular an RIA test (See Clin. Exp. Immunol., 1984 (57); 600-608. Liver-kidney microsomal auto-antibodies are detected by radioimmunoassay and their relation to anti-mitochondial antibodies in inflammatory liver diseases), which h the general disadvantages of RIA tests (especially difficulty of obtaining and handling labelled reagents).
A diagnostic test for AIH has been described which utilizes a fusion protein comprising cytochrome P450 (Gueguan et al., 1989, Biochem. Biophys. Res. Comm., 159 (2), 542-547). A human cytochrome P450 peptide fragment containing an imnmunodominant epitope of cytochrome P450 IID6 binds specifically with the anti-LKM autoantibodies produced during autoimmune hepatitis, and this peptide is the basis for a diagnostic test described in U.S. Pat. No. 5,830,667 issued to F. Alvarez, Nov. 3, 1998. Such an antigen, however, has the disadvantage of also bringing about false positives, especially because of the presence of the associated protein. Thus,
Guo Linsheng
Mason Andrew L.
Xu Lizhe
Alton Ochsner Medical Foundation
Epps-Ford Janet L.
Jones Day
McGarry Sean
LandOfFree
Identification of a novel retrovirus associated with primary... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Identification of a novel retrovirus associated with primary..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Identification of a novel retrovirus associated with primary... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3191085