[A case report of ulcerative colitis complicated with protein losing enteropathy and colon cancer in a young female].
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Biomedical subjects
Publications and source records attributed to K Tsuchimoto.
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The correlation between the histological features of liver biopsy specimens before interferon (IFN) treatment and the clinical effect of IFN administration on chronic hepatitis C was investigated. A study of the relation between several histological features that were graded in 60 liver biopsy specimens from chronic hepatitis C patients before IFN treatment disclosed that the grade of portal fibrosis was positively correlated with the grade of other inflammatory features, including piecemeal necrosis and portal and lobular inflammation. The degree of portal fibrosis adversely affected the rate of normalization of ALT levels in chronic hepatitis C during and after IFN treatment. We reexamined 36 liver biopsy specimens that showed a moderate degree of portal fibrosis, and found that the degree of piecemeal necrosis was inversely correlated with the extent of lymphoid follicle formation in the portal tracts. During IFN therapy, the group of chronic hepatitis C patients who showed marked piecemeal necrosis and less lymphoid follicle formation in the liver specimens had a poor response to IFN treatment, whereas another group that showed marked lymphoid follicle formation and little piecemeal necrosis in the liver specimens had a good response to IFN. These relationships gradually disappeared after the completion of IFN treatment.
A gene encoding the variable regions of the heavy and light chains of a mouse monoclonal antibody designated H2, which specifically reacts with human liver cells, was cloned into a phagemid vector. The clone of the variable region was designed to be expressed as a separate protein, the structure of which is the same as that of the mouse antibody. The cloned phage protein specifically reacted with anti-idiotypic antibodies produced in rabbits against the original mouse antibody, and this reaction was specifically blocked by the original antibody. The soluble protein, expressed as a fusion protein, was detected as a single 30-kd band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and specifically bound to an anti-H2 idiotypic antibody as determined by Western blot analysis. Sera of patients with various diseases were assayed for antibodies to anti-H2 by sandwich enzyme-linked immunosorbent assay (ELISA). Only sera from patients with chronic liver disease reacted strongly. This binding was specifically blocked by the cloned soluble protein. The nucleotide sequences of the variable regions were determined by the dideoxy chain-termination method, and the sequences were approximately 95% identical to those of other mouse immunoglobulins. These findings suggest that a human antibody with the same idiotype as a mouse monoclonal antibody that reacts with human liver cells, can be detected in patients with chronic liver disease, suggesting that autoimmunity may be partly responsible for these diseases.
The efficacy of two different high-dose treatment of IFN-alpha 2b was evaluated in this study. Serum hepatitis C virus (HCV) RNA levels were semi-quantified by simplified reverse transcription-polymerase chain reaction. Seventy-one patients with chronic hepatitis C received 10 million units of IFN-alpha 2b daily for 2 weeks or twice a week for 24 weeks. Alanine aminotransferase (ALT) levels overall normalized in 78.1% and 51.6% of the cases at the end of the therapy and 6 months after that, respectively. HCV RNA disappeared in 71.9% and 35.7% of the patients at the end of the therapy and 6 months after that, respectively. There was no significant difference between the 2 different regimes. The efficacy of the treatment was fair in cases in which the pretreatment level of the viral amount was low. The results of this study indicate that daily administration of IFN in the first 2 weeks during 6-month course does not increase the efficacy of the therapy in such a high-dose treatment regime.
This case report describes a case of carcinomatous peritonitis which showed a good response to the combination therapy of 5-FU, leucovorin and cisplatin. A 55-year-old man was admitted with complaints of meteorism due to ascites fluid. Ascitic examination showed a high concentration of CEA and CA19-9, and adenocarcinoma cells. Laboratory and radiological examination did not reveal the origin of the carcinoma, and laparotomy suggested an appendiceal origin. The combination chemotherapy with 5-FU, leucovorin and cisplatin was conducted for a total of 11 courses for 2 years with good QOL until his death. This therapy was suggested to be useful for inoperable carcinomatous peritonitis.
We produced a murine monoclonal antibody (mAb), designated H2-mAb, against a fractionated soluble phase of human liver homogenate which antibody reacted with human liver cells. A human antibody possessing the same idiotype as the H2-mAb, designated LSIA (liver-specific idiotype-bearing antibody), can be measured by a sandwich enzyme-linked immunosorbent assay, using the anti-H2 idiotype antibody. The serum level of LSIA in patients with histologically proven chronic hepatitis (CH) was significantly higher than that in healthy subjects and it was also higher than that in subjects with other diseases, including systemic lupus erythematosus. In a comparison between patients with CH type B and those with CH type C, there was no significant difference in serum levels of LSIA. It was possible to purify LSIA from the sera of patients with CH. The purified LSIA bound to the human cell lines Chang and HCC-M, derived from liver cells and a hepatoma respectively, but not to HeLa cells, a uterine carcinoma derivative. The reactivity of this mAb to HCC-M was weaker than that to Change. Moreover, the presence of LSIA caused an antibody-dependent cell-mediated cytotoxic challenge against Change cells in vitro.
A rare case of Aspergillus aneurysm of the central nervous system (CNS) leading to subarachnoid hemorrhage (SAH) is reported. An 83-year-old woman developed visual disturbance and headache. Computed tomographic scans showed no evidence of aneurysm or tumor in the intracranium. She suddenly died from SAH. Autopsy revealed massive SAH due to ruptured Aspergillus aneurysm of the middle cerebral artery. Aspergillus was suggested to have extended from the paranasal sinuses. Aspergillosis of CNS should be considered in patients with neurological symptoms such as visual disturbance and trigeminal neuralgia, especially in cases of the aged or immunocompromised.
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Conventional RNA extraction methods, such as SDS/phenol and AGPC methods involve time consuming and complicated manipulation procedures, which could be a cause of contamination. In this study, we compared the conventional methods with a method using a newly developed reagent, "SepaGene-RV" to extract RNA. The results obtained by the new method were the same as those obtained by the conventional methods in the comparative experiments with the sera of patients infected with HCV and detection limit tests using recombinant HCV genome. The sensitivity to detect HCV-RNA with SepaGene-RV was equivalent to that of conventionally used methods to extract RNA, and the manipulation procedure was simpler and it was less time consuming. In addition, we could detect plus-stranded and minus-stranded HCV-RNA by polymerase chain reaction assay (PCR) from liver tissue obtained from liver biopsy and peripheral blood mononuclear cells (PBMC). We confirmed that HCV infects not only liver tissue but also PBMC where replication occurs.
We have previously reported the effect of a differentiation inducer, sodium butyrate (SB), on human hepatocellular carcinoma (HCC) cell lines, demonstrating that it was a potent inducer of differentiation. In the present study, we investigated the alteration in expression of an antigen defined by a murine monoclonal antibody, H2, as well as alterations in the expression of other antigens, on the HCC cell lines HCC-T, HCC-M, and PLC/PRF/5, since it is known that specific antigenic changes occur during the differentiation of leukemic cells. The expression of the antigen defined by H2 increased immunocytochemically on HCC-T, HCC-M, and PLC/PRF/5 during treatment with SB. A flowcytometric study showed that almost all the HCC-T and HCC-M cells treated with SB highly expressed this antigen after 5 days' treatment. The antigen expression detected by H2 decreased after the removal of SB from the medium. On the other hand, antigen expression detected by another monoclonal antibody, 5C11, was not changed by this treatment. The expression of intracellular adhesion molecule (ICAM)-1 in HCC-T increased slightly, but that of beta 2-microglobulin and HLA-DR did not change. These results demonstrated that some antigen expression was changed by SB treatment and that the antigen defined by H2 seemed to be highly expressed on human HCC cells in the differentiated state.
We present here a 63-year-old man with chronic hepatitis C who developed bullous lesions and anti-intercellular antibodies when he received interferon (IFN) alpha-2a, 9 MU subcutaneously, 3 times a week for 3 months. The direct immunofluorescence was consistent with pemphigus features. The indirect immunofluorescence showed negative before IFN treatment, but it became positive after 3 months. The immunoblot study showed no specific bands. This is the third report of the bullous lesions with anti-intercellular antibodies following IFN therapy.
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To evaluate the efficacy of a treatment of weekly interferon administration in patients with chronic hepatitis C, 36 patients were randomly assigned to two groups. In one group lymphoblastoid interferon was given at a dose of 6 million units, intramuscularly, once per week for 24 weeks, and no treatment was given to the other. Serum alanine aminotransferase levels in the treated group were significantly lower during therapy than in the control group, although there was no significant difference between these two groups before therapy. The normalization of serum alanine aminotransferase levels at the end of therapy was observed in 50% of the treated group, and in 11.1% of the control group. This difference was statistically significant (p < 0.03). Response to interferon was better in patients with chronic persistent hepatitis or with chronic active hepatitis than in patients with chronic active hepatitis with cirrhosis. Relapse after the end of therapy was observed in 83.3% of the responders. These results indicate that the weekly administration of 6 million units of lymphoblastoid interferon is effective in decreasing serum alanine aminotransferase levels in patients with type C chronic persistent hepatitis or chronic active hepatitis.
Interferon-beta was given weekly for 24 wk, at a dose of 3 million units, intravenously, to 10 patients with chronic hepatitis B who were serologically positive for HBsAg and HBeAg. Their condition was followed for 6 months after the end of therapy. Both serum hepatitis B virus-associated DNA-polymerase activity and alanine aminotransferase level became significantly lower during therapy and during the 6 months after the end of therapy than at the beginning of therapy. In five of 10 patients, the seroconversion from HBeAg positive to anti-HBe positive had occurred by 6 months after the end of therapy, and in four of these five patients, serum alanine aminotransferase level became normal. Weekly interferon-beta administration over 6 months seems effective in inducing seroconversion and in normalizing serum alanine aminotransferase level.
Changes of nucleotide sequences and expressions of cellular oncogenes in human hepatoma cell lines, PLC/PRF/5, HCC-M and HCC-T cells, were examined by Southern and Northern blot analyses. The probes used are DNA fragment of myc, N-, H-, K-ras, fos, fms, raf, erb-A, erb-B, and erb-B2 genes and synthetic oligonucleotides corresponding to the part of N-, H-, K-ras genes. The results are as follows. DNA amplification and rearrangement were not detected in these three human hepatoma cell lines. Point mutations at codons 12, 13, and 61 in N- and K-ras genes were not demonstrated in these cell lines. N-, H-, K-ras and myc transcripts were detected in these three cell lines. However, fos gene transcript was detected only in PLC/PRF/5 and HCC-M cells which were derived from hepatitis B related hepatocellular carcinoma and having integrated hepatitis B virus (HBV) DNA. These data showed that there are no specific proto-oncogene expression into RNA except for myc and ras genes, nor DNA rearrangement in these 3 human hepatoma cell lines with regards to at least 10 different oncogenes examined and suggest the relationship between fos gene expression and integration of HBV DNA in host cell DNA.
Protein from hog which is recognized by human monoclonal antibody (HB4C5), generated from a patient with large cell lung carcinoma, was identified as carboxypeptidase A by comparison of the protein with carboxypeptidase A in enzymatic activity, immunologic reactivity, and amino acid sequence. Carboxypeptidase A activity was also found in human cancer tissue, and purified antigen from cancer tissue recognized by the antibody HB4C5 was reacted with rabbit anti-carboxypeptidase A serum, indicating that carboxypeptidase A is an antigen of HB4C5. Since large amounts of carboxypeptidase A can be obtained from porcine sources, a simple method for its purification was established. The fraction which was most reactive with HB4C5 was obtained from acetone powder of porcine pancreas by successive applications of water extraction, ammonium sulfate precipitation, trypsin treatment, and Mono Q column chromatography. Its apparent molecular weight was 40,000, according to SDS polyacrylamide gel electrophoresis. When the reactivity of IgG in sera with the purified carboxypeptidase A was measured, the detection rates for lung, ovary, larynx, uterus, and liver cancer were more than 50%, while the rates for stomach and breast cancer were around 30%, and pancreatic cancer, benign diseases, and normal controls were minimally detected.
To investigate hepatitis B virus (HBV) biology in vitro, the transfection of recirculized HBV DNA into Chang cell line was performed. Linear HBV DNA was isolated from recombinant HBV DNA, pHBR105, which includes the whole genome of HBV and was recirculized. Chang cells were transfected with this recirculized HBV DNA by the two different procedures of calcium/phosphate coprecipitation and electroporation. After the transfection, the presence of large nucleated cells with multinuclei and ground-glass cytoplasma were noticed and these cells seemed to proliferate faster than untreated Chang cells. Transient expression of hepatitis B virus surface antigen (HBsAg) was demonstrated in cytoplasma of transfected cells by indirect immunofluorescence. HBsAg was not detected in the culture supernatants by radioimmunoassay. The extra-chromosomal HBV DNA was detected in the transfected cells by both procedures 7 weeks after the transfection by Southern blot analysis but it was lost 4 weeks after that. It was demonstrated that it was possible to transfect Chang cells with HBV DNA and that DNA was functioning to express HBsAg transiently.
A human hepatoma cell line, designated HCC-T, was established. The tumor was surgically obtained from a Japanese male patient with hepatocellular carcinoma (HCC) arising in a cirrhotic liver that had supposedly developed from nonAnonB (NANB) chronic hepatitis. HCC-T exhibited a typical morphology of epithelial cells in culture. Population doubling time was 24 hours and HCC-T cells had characteristics of malignant cells demonstrated by the ability to grow in a soft agar medium and transplantability to nude mice. The histologic condition of the tumor transplanted to a nude mouse showed similarity to the original tumor. A chromosome analysis showed that there were ten identifiable marker chromosomes and sex chromosomes with its modal number of 64. Alpha-fetoprotein (AFP) production was demonstrated by direct immunofluorescence study, but albumin or hepatitis B surface antigen were not detectable. The integration of hepatitis B viral DNA was not demonstrable in the genome of HCC-T cells or the original hepatoma.