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In the liver biopsy specimens of all six patients with acute non-A, non-B hepatitis, the lymphocytic interaction with hepatocytes and sinusoidal endothelial cells was observed by electron microscopic study. Lymphocytes were in a close contact with damaged hepatocytes and interrupted endothelial cells, and the microvilli on the surface of these damaged hepatocytes were degenerated and lost. These findings pointed out the possibility that the lymphocyte may play one of the important roles in hepatocytic damage and endothelial cell damage in acute non-A, non-B hepatitis.

The development of hepatocellular carcinoma (HCC) is very closely associated with chronic liver disease. In the present study, the prevalence of the hepatitis B virus (HBV) and hepatitis C virus (HCV) infection as a causative role in the development of HCC was analysed in 253 patients with HCC, who were admitted to our hospital during 1976-90. Among these patients, 68 (27%) were positive for HBsAg but negative for anti-HCV antibody (group I); in contrast, 147 (58%) were negative for HBsAg but positive for anti-HCV antibody (group II), 19 (7.5%) were both positive (group III), and 19 (7.5%) were both negative (group IV). To evaluate the serial changes in the prevalence of HBsAg and anti-HCV antibody, changes in the number of patients were compared between group I and group II. The number of group I patients reached a peak during 1982-84 and was thereafter followed by a decreasing trend, whereas the number of group II patients steadily increased and reached a plateau over 6 recent years. These results suggest that HCV infection recently seems to play a more important role in the development of HCC than chronic HBV infection, even in the Nagasaki Prefecture, where the HBV carrier rate is higher than elsewhere in Japan.

The effects of tumor necrosis factor-alpha (TNF-alpha) on DNA synthesis in AH66 rat hepatoma cells and rat hepatocytes were analysed by means of [3H]thymidine incorporation. DNA synthesis in AH66 cells was suppressed when AH66 cells were directly incubated with TNF-alpha. When primary culture of rat Kupffer cells was incubated with hepatocyte conditioned media pretreated with TNF-alpha (0-200 U/ml), and AH66 cells were then treated with these hepatocyte/Kupffer cell-conditioned media, TNF-alpha used in the pretreatment caused a dose-dependent increase in DNA synthesis in AH66 cells with a maximum effect amounting to a more than 10-fold increase. In contrast, DNA synthesis in primary culture of rat hepatocytes was not stimulated by the TNF-alpha-pretreated hepatocyte/Kupffer cell conditioned media. These results suggest that TNF-alpha-mediated hepatocyte-Kupffer cell interaction selectively promotes proliferation of rat hepatoma cells.

Two patients, a 3-year-old female and a 1-year-old female, both with a focal encephalopathic process associated with influenza A virus infection, are reported. Both children had neuropsychologic signs suggesting frontal and limbic dysfunction, without disturbances of consciousness or motor function, and had good recoveries. The results of single-photon emission computed tomography and electroencephalography support the finding of reversible impairment of the frontal and limbic areas. Focal reversible encephalopathy has rarely been reported in association with influenza virus infection, although it often provokes diffuse encephalopathies, with a poor prognosis.

We analyzed the serial changes in serum pancreatic enzyme activities by transcatheter arterial embolization (TAE) in 20 hepatoma patients with liver cirrhosis in an attempt to evaluate the incidence of the pancreatic tissue damage by TAE. Serum amylase activities increased in two (10%) cases, elastase 1 levels in six (30%) cases, and trypsin and pancreatic secretory trypsin inhibitor (PSTI) levels in each of five (25%) cases. Consequently, TAE resulted in the elevation of at least more than one serum pancreatic enzyme in eight (40%) of 20 cases, although none had clinical symptoms related to pancreatitis. When the adverse effect on the pancreatic tissue was compared among 6 cases of the superselective TAE and 14 cases of the nonsuperselective TAE, which were performed from the segmental and the nonsegmental hepatic arteries, respectively, the elevation of serum pancreatic enzymes was caused only by nonsuperselective TAE, not by superselective TAE. The volumes of Spongel and Lipiodol used or the injected doses of the anticancer agent mitomycin C were not different between the two groups. These results indicate that TAE for the treatment of hepatoma frequently causes pancreatic tissue damage, and the position of the inserted catheter tip is very important to avoid the pancreatic tissue damage by TAE.

Transforming growth factor beta 1 (TGF-beta 1) is known to inhibit hepatocyte growth in vitro and in vivo. In this study, we analyzed the effect of TGF-beta 1 on alpha-fetoprotein (AFP) and albumin gene expression in HuH-7 human hepatoma cells. TGF-beta 1 inhibited cell growth in a dose dependent manner. The cellular secretion rate of AFP but not albumin was suppressed significantly by TGF-beta 1. TGF-beta 1 caused a significant reduction in the level of AFP mRNA. In contrast, the levels of albumin mRNA or beta-actin mRNA were not changed by TGF-beta 1. In transient transfection experiments, TGF-beta 1 resulted in selective repression of AFP promoter activity. These results suggest that TGF-beta 1 is one of the key factors involved in the differential regulation of the AFP gene and the albumin gene.

A rare case of hepatocellular carcinoma who developed the complication of the sign of Leser-Trelat is reported. The patient, a 57-year-old male, visited our hospital with complaints of generalized malaise and anorexia. A diagnosis of hepatocellular carcinoma was made based on elevated alpha-fetoprotein measurement, ultrasonography, and hepatic arteriography findings. Chest x-ray film suggested pulmonary metastases of hepatocellular carcinoma. Thereafter, complications of the seborrheic keratosis developed in the trunk and the skin lesion was diagnosed as the sign of Leser-Trelat associated with hepatocellular carcinoma. The patient died of pneumonia 9 months after development of the sign of Leser-Trelat.

The expression of the alpha-fetoprotein (AFP) gene in hepatocellular carcinoma (HCC) and liver cirrhosis by in situ hybridization analysis of AFP mRNA in cryostat and paraffin-embedded tissue sections was studied. In HCC sections the majority of tumor cells showed positive hybridization with a 3H-labeled AFP complementary DNA probe. The number of radioactive hybrid grains detected in HCC sections generally paralleled the level of serum AFP in the patient. In liver cirrhosis sections a small number of cells showed positive hybridization. These cells were dispersed in the tissue and morphologically indistinguishable from surrounding hepatocytes. Each of these cells contained a high level of hybridization signals to permit easy identification. In situ hybridization analysis of AFP mRNA may be of use in detecting preneoplastic cells in liver cirrhosis that cannot be defined on the morphologic and immunohistochemical basis.

肝細胞癌の早期診断はきわめて難しい.著者らは細小肝細胞癌7例を経験したので,その臨床病理学的特徴につき述べる.7例中6例は診断時自覚症状は軽微であった.臨床所見中変形し増大する肝腫に注目する必要がある.血中AFPは5例が陽性で診断に役立ち,とくに経時的な追跡測定が重要である.特異的診断法ではないが,HBs抗原,アルカリホスファターゼ,総コレステロール,LDH, LAP,トランスアミナーゼ解離が有用な例があった.肝動脈造影は有力な診断法で5例にTumor stainがみられた.AFP陰性,肝動脈造影でTumor stainがみられない2例中1例は腫瘤の吸引細胞診で診断がついた.組織型は全例索状型であった.非癌部組織は全例肝硬変または肝線維症で,肝細胞癌の治療上の問題点につき述べた.術後4年生存している1例では,肝シンチグラム,肝動脈造影で切除部は修復されており,肝硬変でも再生するこ とを示していると考えられた.

AFP産生肝細胞癌の局在診断に抗AFP抗体の利用を企て基礎実験を行った.すなわち呑龍ラットの大腿皮下にAFP産生腹水肝癌AH-7974細胞を移植し作製した腫瘍に放射性ヨード標識抗ラットAFPウマ抗体を静注し,以下の実験結果を得た.なおコントロールとして放射性ヨード標識正常ウマIgGを用いた.(1)全身シンチグラムで腫瘍は投与48時間後からhot areaとして認められ,120, 168時間目に明瞭となった.(2)組織(cpm/g)/血液(cpm/g)放射活性比をみると腫瘍は7日目にコントロールに比し4倍の高値を示した.各臓器中脾,腎は比較的高かった.(3)腫瘍の細胞下分画の放射活性は細胞膜,核分画で最も高かった. (4)腫瘍組織の7日目のオートラジオグラムで放射活性を示すgrain集合像は細胞表面に存在した.以上のごとく抗AFP抗体はAFP産生腫瘍と特異的に結合すること,同時にシンチグラムによる局在診断が可能であることが分った.

Two antibodies were prepared for use in a sandwich enzyme immunoassay of human IgG. Completely purified guinea pig anti-human IgG was labelled with beta-D-galactosidase (EC 3.2.1.23), using a heterobifunctional cross-linker named GMBS. Partially purified anti-human IgG was immobilized on a new solid support: Amino-Dylark balls. Optimal conditions for immobilizing the antibody, using glutaraldehyde as the coupling reagent, were studied in detail. With the enzyme-labelled antibody and the solid-phase anti-human IgG, a sandwich enzyme immunoassay of human IgG with a lower limit of detection at 10.5 pM (0.3 ng/tube) was developed. A comparative study of the EIA method and a laser nephelometric method showed a good correlation. The specificity of the assay was excellent: all 4 types of IgG tested showed the maximum 0.0001%; human IgA, IgM and albumin possessed the maximum 0.54% in their cross-reactivity values with human IgG.

Immunoliposomes, bearing Fab' fragments of anti-carcinoembryonic antigen (anti-CEA) monoclonal antibody, have been prepared according to a new method developed in our laboratory. First, egg phosphatidylcholine liposomes were coated with pullulan derivatives. Then, SH-bearing Fab' fragments of anti-CEA were conjugated to the pullulan derivatives attached to the liposomal surface. These immunoliposomes showed specific binding to CEA-producing cells, BM314, which was approximately 20-fold greater than to CEA-non- producing cells, RPMI #4788. In addition, these immunoliposomes were endo cytosed by colon cancer cells, which was confirmed by fluorescence microscopy using liposomes labelled by a hydrophobic fluorescent probe, terbium trisace tylacetonate.

RIAによるHBs抗原測定は鋭敏であるが,RI使用のため種々の制約がある.そこでRIAと同感度をもつといわれるEIAに着目した.標識酵素にβ-D-gal,吸着固相にSilicon rodsまたはGlass beadsを用い検討した.その結果,(1)精製HBs抗原の定量曲線から最小検出濃度は1ng/mlでRIAと同感度を示し,両者の相関はr=0.99と高い.(2)RIAでHBs抗原陽性の37検体につきEIAとの相関をみるとr=0.87と高い.(3)抗体IgGを化学的に吸着させたGlass beads使用の方が自然吸着させたSilicon rodsよりincubationが短時間で,血清添加の影響も少なかった.(4)フィールドにおける臨床試用を行うと大学病院看護婦92名はEIAで陰性を示しRIAと一致した.一般職員451名中11名がEIAで陽性,うち10名はRIAと一致した.残り1名はRIA陰性のHBs抗原陽性者と思われた.肝疾患多発地域住民743名中39名(5.2%)がEIAで陽性を示した.以上からEIAはRIAに代りうる測定法であることが分った.

清酒換算毎日3合(80g)以上,期間10年以上の常飲者34例をA群(肝硬変・前硬変,17例),B群(非硬変,17例、に分け臨床病理的に検討した.臨床的事項:理学的に肝腫,黄疸,クモ状血管腫,腹水ともA群に圧倒的に多い.肝機能ではBSP, TTT, ZST, AL-PともにA群に異常例が多く又,A群でGOT>GPTの傾向がある.飲酒量・持続期間と肝組織像との比較では清酒換算8合(200g)以上,期間15年以上の常飲者にA群に入る例が多い.病理的事項:電顕的に肝細胞胞体基質内に160Aのperiodicityを有す「あみの目状」の結晶構造がみられたことは注目される.また,経時的肝生検により,徐々に進行し最終的に壊死後性肝硬変の完成をみた例を経験したが,アルコールによる壊死後性肝硬変の成立に示唆を与えるものと考えられる.

われわれは, 25才の初産婦で,分娩中に高熱,嘔吐,下痢および全身皮膚の発赤が突然出現し,その後ショック状態に陥った症例を経験した.細菌学的検査では,患者の腟分泌物から黄色ブドウ球菌が検出され,さらに, toxic shock syndrome toxin-1 (TSST-1)を産生することも証明された.以上より,本症例をtoxic shock syndromeと確定診断した.米国においては,すでに2000例以上が報告されているが,本邦での報告ははなはだ少ない.本邦において,膣分泌物から病原菌並びにTSST-1が証明されたのは,本例が第1例であるので報告する.

3回の摘出術と2回の肝動脈塞栓術(TAE)により10年8カ月生存している肝細胞癌の1例を報告した.症例は62歳の女性,1971年肝生検で肝硬変と診断.1973年10月血中AFP上昇のため当科入院.HBs抗原,HBe抗原ともに陽性.血管造影で腫瘍膿染像をみとめ,摘出術施行,右葉後上区域の3.5cm大の肝癌を摘出した.非癌部は乙型肝硬変であった.6年10カ月後に肝癌の再発がみられ,やはり右葉後上区域にて2.0cm大の肝癌を摘出.さらに10カ月後に肝癌が出現,右葉後下区域より1.0cm大の肝癌を摘出した.組織学的には,3回ともtrabeculartypeであった.さらに1年8カ月後肝癌が出現,TAEを2回施行し外来通院中である.本例の肝癌発生様式は,多中心性と思われるが,血中AFPの厳重なfollowにより3回もの肝癌摘出術に成功し,10年以上の生存をみているため報告した.

Highly purified polyclonal or monoclonal antibodies to human or rat α-fetoprotein (AFP) were proved to be cytotoxic to AFP-producing hepatoma cells. Rat with transplanted hepatoma lived longer by the antibody treatment. The polyclonal horse antibody to human AFP was administered to 25 cases of patients mostly with hepatoma. In 7 out of 25 cases, serum AFP levels were markedly suppressed for a long period. Daunomycin, an antitumor drug, was conjugated to the antibodies according to the method of E. Hurwitz and M. Sela. The conjugate revealed a striking antitumor effect compared to antibody or drug alone. Toxic effect of daunomycin upon the host animals was markedly decreased by the conjugation with antibody protein. Any serious side effect of administration of horse antibody was observed in the patients.

The tumor imaging using radiolabeled antobodies to α-Fetoprotein (AFP) was undertaken in 27 patients with various AFP producing tumors. In 7 of 15 patients (47%), positive images were obtained using polyclonal antibody and 4 positive and 3 doubtful images were obtained in 12 patients using 4 monoclonal antibodies. The tumor/serum ratios of AFP showed high levels in positive cases, and all monoclonal antibodies could bind to AFP in 7 patients with various primary tumor sites in vitro.

To determine the diagnostic value of anti-HBc IgM in acute viral hepatitis or chronic liver disease B, Anti-HBc IgM was measured by a RIA and an ELISA in 32 patients with acute hepatitis (4 with type A, 15 with type B and 13 with type non A non B), 18 patients with chronic hepatitis and 19 patients with liver cirrhosis. In acute hepatitis B, anti-HBc IgM (both RIA and ELISA) was positive in 14(93%) of 15 patients and its cut-off index value was very high. However, anti-HBc IgM was always negative in one patient with typical course of type B. In 1 of 4 patients with acute hepatitis A and 2 of 13 with non A non B, anti-HBc IgM (RIA and/or ELISA) was positive. These 3 patients were positive for anti-HBs at the onset of disease, so we could not made the diagnosis of acute hepatitis B. Anti-HBc IgM was positive in 21(51%) of 37 patients with HBsAg-positive chronic liver disease by RIA and in 11 (30%) by ELISA, and its cut-off index value was relatively low. These results suggest that when adequate cut-off index value is established, anti-HBc analysis is useful for differentiating recent and current infections from remote infections.

This study reports the use of radiolabeled antibody to AFP and CEA for the detection and localization of AFP- or CEA-producing tumors. Thirty-one patients received 131I-labeled anti-AFP or anti-CEA antibodies. Photoscans were taken at 24 and 48 hours after injection of radioantibodies. In three of six patients with CEA-producing tumors, radioimmunodetection with anti-CEA antibody showed positive scans. In AFP-producing tumors, 7 of 15 patients had positive findings on immunoscintigraphy using polyclonal anti-AFP antibody, and two of nine patients had positive findings when monoclonal antibodies were used. Analysis of radioantibody in the blood after injection showed both complex and free antibody with immunoreactivity in the circulation, and smaller complexes were seen to form after administration of monoclonal antibodies.

Human of rat alpha-foetoprotein (AFP) was highly purified from ascitic fluid or serum of hepatoma bearers. The purification was carried out mainly by means of immunoadsorbent chromatography using Sepharose coupled to specific anti-AFP antibody with BrCN activation, and by Sephadex gel filtration. Horses were immunized with the purified AFP and the specific antibody was isolated from the antiserum by means of an immunoadsorbent coupled to purified AFP. The specific antibody was found to bind specifically with AFP-producing tumour cells. The antibody was applied for radio immunodetection of the tumour. 125I-labelled antibody was administered to patients or rats with hepatoma, and radioactivity localized in the tumours was scintiscanned with a scintillation camera. In this way, the location of the tumour was detected in about 50% of the hepatoma bearers. The cytotoxicity of the antibody was clearly demonstrated both in vitro and in vivo in animal experiments. The antibody was administered to patients with advanced hepatoma. Although no improvement of the disease was demonstrated, serum AFP levels decreased greatly and in some cases low AFP levels were maintained for long periods suggesting that the antibody suppressed AFP-producing hepatoma cells. No significant side effects were observed in patients who had been administered with the horse antibody.

The sugar-chain structures of circulating alpha-fetoprotein in patients with hepatocellular carcinomas differ from those in patients with cirrhosis. We studied the reactivity of alpha-fetoprotein with two lectins, Lens culinaris agglutinin A and erythroagglutinating phytohemagglutinin, to monitor the evolution of hepatocellular carcinoma in patients with cirrhosis.Among 361 patients with cirrhosis caused mainly by chronic hepatitis B or hepatitis C virus infection, 33 with base-line serum alpha-fetoprotein concentrationsor = 30 ng per milliliter or more were found to have hepatocellular carcinomas during a mean follow-up of 35 months. The lectin-reactive profiles of the alpha-fetoprotein in the serum of these 33 patients were analyzed and compared with those in the serum of 32 patients with cirrhosis who had increased base-line serum alpha-fetoprotein concentrations and were followed for at least 24 months but in whom hepatocellular carcinoma did not develop.At the time of tumor detection, 24 (73 percent) of the 33 patients with cirrhosis and hepatocellular carcinoma had higher percentages of L. culinaris agglutinin A-reactive alpha-fetoprotein (alpha-fetoprotein L3), erythroagglutinating phytohemagglutinin-reactive alpha-fetoprotein (alpha-fetoprotein P4+P5), or both than the 32 patients with cirrhosis but no hepatocellular carcinoma. Among the 24 patients, one or both of the markers were first elevated 3 to 18 months before the hepatocellular carcinoma was detected by imaging techniques.Measurements of the alpha-fetoprotein L3 and alpha-fetoprotein P4+P5 fractions of serum alpha-fetoprotein allow the differentiation of hepatocellular carcinoma from cirrhosis in some cases and serve as predictive markers for the development of hepatocellular carcinoma during the follow-up of patients with cirrhosis.