Expression of the SART1 tumor-rejection antigen in hepatocellular carcinomas.

We previously reported a tumor-rejection antigen, SART1259, possessing tumor epitopes capable of inducing cytotoxic T lymphocytes (CTL) in epithelial cancer patients. The present study investigated the expression of the SART1259 antigen in hepatocellular carcinomas (HCC) in order to explore for a potential molecule for use in specific immunotherapy of HCC patients. Expression of the SART1 antigens in samples was analyzed by western blot analysis with anti-SART1259 and anti-SART1800 polyclonal antibodies. In addition HLA-A24- restricted CTLs were induced from peripheral blood mononuclear cells (PBMCs) of HLA-A24+ HCC patients by the SART1690-698 (EYRGFTQDF) peptide with an HLA-A24 binding motif. The SART1259 antigen was detected in the cytoplasmic fraction of 6 of 8 HCC cell lines and 12 of 23 (52%) HCC tissues, but in none of the normal liver tissues or those of chronic hepatitis or cirrhosis. The HLA-A24 restricted and SART1-specific CTLs recognized the HLA-A24+ and SART1259+ HCC cells. Further, in peripheral blood mononuclear cells of HCC patients, the SART1690-698 peptide induced CTLs that reacted to the HCC cells in an HLA-A24-restricted manner. These results suggest that the SART1259 antigen could be an appropriate target molecule for use in specific immunotherapy of HCC patients.

Expression of the SART3 tumor rejection antigen in renal cell carcinoma.

PURPOSE: We recently reported that SART3 tumor rejection antigen is recognized by HLA class I restricted cytotoxic T lymphocytes from patients with esophageal cancer. We now investigate the expression of SART3 antigen in renal cell carcinoma to identify an appropriate molecule that may be used in specific immunotherapy of renal cell carcinoma. MATERIALS AND METHODS: Renal cell carcinoma and nontumorous kidney tissues were obtained at surgery. A section of each sample was minced with scissors and stored at -80C until use. SART3 antigen expression was examined in uncultured renal cell carcinoma and nontumorous kidney tissues. We also evaluated the ability of derived peptides to include cytotoxic T lymphocytes in peripheral blood mononuclear cells from patients with renal cell carcinoma. RESULTS: The SART3 antigen was detected in all renal cell carcinoma cell lines, primary cultures of renal cell carcinoma and nontumorous kidney tissues, and in the cytosol of 57% and 15% of renal cell carcinoma and nontumorous kidney tissues, respectively. HLA-A2402 restricted and tumor specific cytotoxic T lymphocytes (KE4) used in cloning of the SART3 gene were significantly cytotoxic to cells from renal cell carcinoma cell lines and primary cultures of renal cell carcinoma tissue but they did not lyse normal cells, including those from primary cultures of nontumorous kidney tissue. The SART3 peptides derived from positions 109-118 and 315-323 induced HLA-A24 restricted cytotoxic T lymphocytes to renal cell carcinoma cells from peripheral blood mononuclear cells of patients with renal cell carcinoma. CONCLUSIONS: The SART3 antigen and derived peptides may be applied to the specific immunotherapy of HLA-A24+ renal cell carcinoma.

Expression of the SART1 tumor rejection antigen in renal cell carcinoma.

We have previously described the SART1 gene, which encodes both the SART1(259) antigen expressed in the cytosol of the majority of squamous cell carcinomas and some adenocarcinomas and the SART1(800) antigen expressed in the nucleus of the majority of proliferating cells. The SART1(259) antigen is recognized by HLA-A24 and A26-restricted cytotoxic T lymphocytes (CTLs). The present study investigated the expression of these two antigens in renal cell carcinomas (RCCs) in order to identify an appropriate molecule for use in specific immunotherapy for RCC patients. These two antigens were detected in all RCC cell lines and cells of the primary cultures of the RCCs tested. Further, they were detectable in cells of the primary cultures of non-tumorous kidney tissues. In contrast to these cultured cells, SART1(259) was detectable in only a few uncultured RCC tissues (5/20, 25%) and was undetectable in non-tumorous kidney tissues. SART1(800) was also scarcely detectable in uncultured RCC tissues (3/20, 15%) and non-tumorous kidney tissues (4/20, 20%). HLA-A2402-restricted and tumor-specific CTL (KE4-CTL) used for the cloning of the SART1 gene showed significant levels of cytotoxicity to both the cells from the RCC cell line and the cells from the primary cultures of RCC tissues, but did not lyse any normal cells, including cells from the primary cultures of non-tumorous kidney tissues. The SART1-derived peptide at positions 690-698 induced HLA-A24 restricted CTLs cytotoxic to RCC cells from peripheral blood mononuclear cells (PBMCs) of RCC patients. Therefore, the SART1 peptide could be an appropriate molecule for use in peptide-based specific immunotherapy for RCC patients.

Expression of the SART1 tumor-rejection antigen in human osteosarcomas.

We recently reported a tumor-rejection antigen, SART1259, possessing tumor epitopes capable of inducing cytotoxic T lymphocytes (CTLs). This study investigated the expression of SART1259 antigen in osteosarcoma and other skeletal malignant tumors to explore for a potential molecule for use in specific immunotherapy. The SART1259 antigen was detected in the cytosol fraction of 13 of 21 (62%) osteosarcoma cell lines and 3 of 8 (38%) osteosarcoma tissues, and 3 of 10 (30%) malignant fibrous histiocytoma (MFH) tissues. The HLA-A24+ and SART1259+ osteosarcoma cells were recognized by the HLA-A24 restricted and SART1 specific CTLs. These results raise a possibility that the SART1259 would be an appropriate molecule for use in specific immunotherapy of approximately one-third of HLA-A24+ patients with osteosarcoma and MFH.

A simultaneous monitoring of Lens culinaris agglutinin A-reactive alpha-fetoprotein and des-gamma-carboxy prothrombin as an early diagnosis of hepatocellular carcinoma in the follow-up of cirrhotic patients.

To elucidate the risk factors for developing hepatocellular carcinoma (HCC) during the follow-up of patients with liver cirrhosis (LC), outpatients with LC were examined periodically by means of serum biochemical assessments, ultrasonography, and computed tomography. Risk factors for HCC were statistically analyzed. We also examined an efficacy of Lens culinaris agglutinin A-reactive profiles of alpha-fetoprotein (AFP-L3%) and des-gamma-carboxy prothrombin (DCP) value using a highly sensitive DCP determination kit (ED036) for the early recognition of HCC. The AFP-L3% and the ED036 value were retrospectively determined with stored serum samples. HCC was diagnosed in 21 of the 78 patients with LC during the follow-up period (mean follow-up period: 42 months). The estimated cumulative incidence of HCC was 25% with 3 years and 48% with 5 years. The most significant risk factor for the development of HCC in LC patients was found to be the mean serum AFP concentration from the year before the HCC detection (p=0.02). At the time of the recognition of HCC, the positive rates of the tumor markers were: serum AFP concentration 14%, serum DCP value 5%, AFP-L3% was 33%, and that of ED036 43%. The positive rate in collaborative use of AFP-L3% and ED036 was 67%. The simultaneous determination of the AFP-L3% and the ED036 value was shown to be effective for the early detection of HCC.

Expression of the tumor-rejection antigen SART1 in brain tumors.

We have reported a tumor-rejection antigen, SART1(259), possessing tumor epitopes capable of inducing cytotoxic T lymphocytes (CTLs) in epithelial-cancer patients. This study investigated the expression of SART1(259) antigen in brain tumors, to explore for a potential molecule for use in specific immunotherapy of patients with brain tumors. The SART1(259) antigen was detected in the cytosol fraction of 13 of 18 (72%) glioma cell lines and in 12 of 34 (35%) brain-tumor tissues, with a higher rate of expression among malignant gliomas (5/10, 50%) and schwannomas (3/4). HLA-A24-restricted and SART1-specific CTLs recognized the HLA-A24+ and SART1(259)+ glioma cells, and the levels of recognition correlated both with HLA-A24-antigen expression level and with the concentration of the SART1 peptide antigen. Therefore, the SART1(259) antigen could be a target molecule for specific immunotherapy of patients with brain tumors expressing HLA-class-1 antigens.

Detection of skewed X-inactivation in two female carriers of vasopressin type 2 receptor gene mutation.

Most cases of congenital nephrogenic diabetes insipidus (NDI) are inherited in an X-linked manner, which is due to the mutations of the vasopressin type 2 receptor (V2R) gene. However, recent reports have presented female NDI patients with heterozygote V2R gene mutations. The mechanism of inheritance was thought to be skewed X-inactivation. We present a family with congenital NDI. Three male members were diagnosed with NDI, and examination of their V2R gene revealed a G inserted at nucleotide 804 of the open reading frame. Three female individuals display different degrees of symptoms of NDI, and all of them possess both the normal and abnormal genes. The X-inactivation patterns of the female members were investigated via the detection of methylated trinucleotide repeat in the human androgen receptor gene. The grandmother showed extremely skewed methylation of one X chromosome, and the mother revealed moderately skewed methylation. The daughter of the grandmother's sister, who has no symptoms of NDI, showed random methylation. The highly skewed X-inactivation pattern of the grandmother suggests that her NDI phenotype is caused by dominant methylation of the normal allele of V2R gene.

CD4+ hepatic cancer-specific cytotoxic T lymphocytes in patients with hepatocellular carcinoma.

We investigated T cell immunity against hepatocellular carcinoma (HCC), and showed that both peripheral blood mononuclear cells and tumor-infiltrating lymphocytes incubated with interleukin-2 alone displayed HLA-nonrestricted but hepatic cancer-specific cytotoxicity in a majority of patients with HCC. Namely, they lysed both HCC and cholangiocellular carcinomas in an HLA-nonrestricted manner, but they did not lyse any tumors with the other histological types tested, normal hepatocytes, or the cells transfected with hepatitis C virus or MUC1 gene. These CTL lines and clones were phenotypically CD3+CD4+CD8-. These unique CTL could play important roles in T cell immunity against HCC.

Carboplatin as an anticancer agent for transcatheter arterial chemoembolization in patients with hepatocellular carcinoma.

Transcatheter arterial chemoembolization (TACE) is a conservative treatment in patients with hepatocellular carcinoma (HCC). In the present study, 30 patients with unresectable HCC underwent TACE using carboplatin (300 mg), and their clinical results were evaluated. After TACE, 18 (60.0%) of 30 patients demonstrated tumor size reduction rates > or = 50%. Of 23 patients with pretreatment serum alpha-fetoprotein (AFP) levels >20 ng/ml (cutoff), 14 (60.9%) showed AFP reduction > or = 75%. The 1-, 2-, 3- and 4-year survival rates were 82.9, 68.1, 45.1 and 37.6%, respectively. The median survival was 2.3 years. The only notable adverse reaction accompanying TACE was a transient myelosuppression. Carboplatin is thought to be a useful anticancer agent in patients with HCC treated with TACE.

[Studies on Rehmanniae radix. III. The relation between changes of constituents and improvable effects on hemorheology with the processing of roots of Rehmannia glutinosa].

The relation between constituents and effects on hemorheology with processing of root of Rehmannia glutinosa was investigated. With the processing to dried or steamed root from crude root, the content of stachyose, which was the highest component in crude root, was decreased and the content of manninotriose was increased. Iridoid glycoside, catalpol, was gradually decreased with the processing. From these results, the content of carbohydrates and catalpol made it possible to estimate the quality of Rehmanniae Radix by processings. On the other hand, the change of pharmacological activities with the processing was investigated. The 50% ethanolic extract (BJ-ext) from steamed root of R. glutinosa increased erythrocyte deformability and erythrocyte ATP contents, inhibited polybrene-induced erythrocyte aggregation, and promoted an activity of fibrinolytic system. The extracts from crude or dried root had weak or no effect. The crude or dried root and steamed root of R. glutinosa showed the different pharmacological activities, and the quality of Rehamanniae Radix by processings may be estimated by investigation of correlation between the changes of constituents and improvable effects of hemorheology.

A sex reversal infant with XX karyotype and complete male external genitalia.

The unusual case of a Japanese newborn XX male is presented. Examination of chromosomes in amniotic fluid cells had shown a normal female karyotype (46,XX), but ultrasonography revealed a penis and a scrotum. The neonate had normal male external genitalia, and serum levels of luteinizing hormone, follicle stimulating hormone, and testosterone were all within the normal range. High resonance chromosome analysis revealed an excess portion on the short arm of one of the X chromosomes. We examined his genomic DNA by polymerase chain reaction (PCR) and detected two Y specific regions in his genomic DNA, the sex-determining region Y (SRY) and pseudoautosomal boundary Y. Nucleotide sequencing of the PCR products of SRY indicated no mutation. These findings suggested that the translocation or insertion of an SRY region on the X chromosome led to the development of testicles and a male phenotype.

[Neoadjuvant chemotherapy of liver tumors metastasized by sigmoid colon cancer: a case report of CDDP/5-FU intraarterial infusion therapy followed by hepatectomy].

A case of liver tumors metastasized by Sigmoid colon cancer was treated with intrahepatic arterial infusion of CDDP (10 mg) and 5-FU (250 mg) via subcutaneously implanted reservoir. After 20 administrations of CDDP and 5-FU, the tumors in the right lobe of the liver disappeared, but those in the left lobe remained. Therefore left hepatectomy was carried out. Six months after the hepatectomy, a nodule recurrence was disclosed in the right lobe of the liver. With re-elevated serum CEA level, subsequent intrahepatic arterial infusion of CDDP and 5-FU the nodule size reduced with normalization of the serum CEA level.

Familial congenital hypopituitarism with central diabetes insipidus.

Congenital hypopituitarism (CH) presenting with central diabetes insipidus is typically associated with midline facial deformities or ophthalmological abnormalities. We present three brothers with CH and central diabetes insipidus not associated with any of these predisposing conditions. All three subjects presented with clinical features typical for CH (neonatal hypoglycemia, short stature, protruding forehead, and microgenitalia). All had hypoplastic genitalia indicating in utero gonadotropin deficiency, and all had complete GH deficiency. One represented low levels of thyroid hormones and TSH, indicating central hypothyroidism. Water deprivation examination in two of the brothers demonstrated complete arginine vasopressin deficiency in one and partial deficiency in the other. Magnetic resonance imaging indicated absence of the pituitary stalk, severe hypoplastic anterior pituitary in all three brothers, and absence of any posterior pituitary gland in two of the three. The other sibling had an ectopic posterior pituitary. This first report of familial CH with central diabetes insipidus may represent a previously unknown midline anomaly and provide new insights into the genetic control of pituitary and hypothalamic development.

Relationship between serum levels of methylguanidine and glycemic control in IDDM children.

OBJECTIVE: To examine the serum levels of methylguanidine in IDDM children and compare them with markers for glycemic control. Reports have indicated that active oxygen, which damages various tissues, increases in diabetes mellitus. The increase of active oxygen is one of the risk factors for diabetic complications. The synthesis of methylguanidine, a metabolic product of guanidine, is mainly regulated by active oxygen. RESEARCH DESIGN AND METHODS: Forty-eight children with IDDM (mean age 13.3 yr) and 17 age-matched nondiabetic control subjects were studied. Diabetic children were divided into a well-controlled group (HbA1c < 8%, n = 24) and a poorly controlled group (HbA1c > 8%, n = 24). Serum concentrations of methylguanidine were measured by enzymatic assay. RESULTS: Levels of methylguanidine in the poorly controlled group (1.31 +/- 0.08 microM) were significantly higher than those in both the well-controlled group (0.85 +/- 0.08 microM) and the control group (0.59 +/- 0.11 microM), respectively (P < 0.01). Methylguanidine levels showed a positive correlation with the levels of HbA1c (P < 0.01) or fructosamine (P < 0.01). No significant correlations were noted between methylguanidine levels and age, sex, duration of diabetes, or insulin dose. CONCLUSIONS: Our data indicate that the levels of methylguanidine in IDDM children might be affected by glycemic control and that the determination of serum methylguanidine levels could be a useful test for evaluating the state of diabetic control.

[Cancer mortality of male workers exposed to benzidine and/or beta-naphthylamine].

The risk of cancer was analysed in a cohort of 604 male workers who had been engaged in manufacturing and/or handling benzidine and/or beta-naphthylamine during the period 1945-71 at two factories located in the city of Osaka. The cohort was followed up from 1 January 1970 to the date of death or 31 December 1986. The mean follow-up time was 16.1 years. A total of 84 deaths was found compared with 112.66 expected based on the mortality of the Osaka population. Thirty-six were found to be dead of malignant neoplasms; 9 stomach, 2 colon, 1 rectum, 3 liver, 1 bile duct, 1 pancreas, 1 maxillary sinus, 6 lung, 8 bladder, and 2 ureter neoplasms as well as 1 case of myeloid leukemia. Seven cases were ascertained on death certificates as neoplasms of uncertain behaviour, all of which were tumors of genitourinary organs except for one case of brain tumor. Cancers of genitourinary organs and tumors of uncertain behaviour showed statistically significant increased standardized ratios (SMR = 12.20, 4.89). The mean age of death of those having genitourinary organs including cancer was 59 years old, and the latent period between the first exposure and the occurrence of the disease was 19.7 years on average. Non-significant increased risks of cancers of the colon, rectum, liver and lung were observed among the workers exposed to benzidine. Among the 7 histologically confirmed cases of these cancers, there were 2 adenocarcinomas of the lung, 1 adenocarcinoma of the rectum, 2 hepatocellular carcinoma and 1 adenocarcinoma of the bile duct. Seven patients with genitourinary tumors were found to have died of other primary cancers.