A case of gastric cancer following living donor liver transplantation.

Only a few cases of de novo malignancy, especially gastric cancer after living donor liver transplantation (LDLT), have been reported. We report a case of gastric cancer following LDLT, after which immunosuppressants were minimized in accordance with the results of the mixed lymphocyte reaction (MLR) assay. A 65-year-old woman had previously undergone LDLT for hepatocellular carcinoma associated with hepatitis B virus infection. The liver graft had been donated by her son. During the course of postoperative surveillance with the MLR assay in order to minimize immunosuppressants, she was incidentally found to have gastric cancer during an endoscopic examination, 8 years after the liver transplantation. She underwent total gastrectomy with lymph node dissection. In this case, gastric cancer was detected 8 years after LDLT, which is longer than previously reported intervals between LDLT and malignancy detection. The number of patients undergoing LDLT is increasing, and the prognosis after liver transplantation has improved. Therefore, endoscopic surveillance programs are important for detecting malignancies in the early stages in liver transplant recipients.

A novel anti-MUC1 antibody against the MUC1 cytoplasmic tail domain: use in sensitive identification of poorly differentiated cells in adenocarcinoma of the stomach.

BACKGROUND: Isolated cancer cells of non-solid type poorly differentiated adenocarcinoma (por2) or signet-ring cell carcinoma (sig) are frequently seen in scirrhous gastric cancers with a very poor prognosis. These cells are often scattered in granulation tissue or desmoplastic fibrotic tissue and tend to be overlooked in routine pathological examination. We aimed to raise a novel antibody that can identify the isolated cancer cells easily. METHODS: Because the MUC1 cytoplasmic tail domain (CTD) has many biological roles including tumor progression and cell adhesion disturbance and is expected to be expressed in isolated cancer cells, we raised a novel monoclonal antibody (MAb) MUC1-014E against an intracellular nonrepeating 19-amino-acid sequence (RYVPPSSTDRSPYEKVSAG: N-1217-1235-C) of the MUC1 CTD, using a synthetic peptide including the 7-amino-acid epitope (STDRSPY: N-1223-1229-C). RESULTS: In the immunohistochemical staining of 107 gastrectomy specimens including 48 por2 and 31 sig lesions, the MAb MUC1-014E showed high rates of positive staining (≥5% of carcinoma cells stained) for por2 (100%) and sig (97%), and of the highest intensity staining (4+, ≥75% of carcinoma cells stained) for por2 (100%) and sig (90%). In the 89 biopsy specimens including 82 por2 and 38 sig lesions, the MAb MUC1-014E showed high rates of positive staining for por2 (100%) and sig (100%) and of 4+ staining for por2 (87%) and sig (84%). All the rates were significantly higher than those with cytokeratins (AE1/AE3 or CAM5.2). CONCLUSIONS: The MAb MUC1-014E is very useful for accurate detection of isolated cancer cells in scirrhous gastric cancers.

Identification of HLA-A*2402-restricted epitope peptide derived from ERas oncogene expressed in human scirrhous gastric cancer.

ERas is a recently identified oncogene involved in the tumorgenic growth of embryonic stem cells. We examined the significance of ERas expression in scirrhous gastric carcinoma, and the possibility of ERas as a tumor-associated antigen of gastric cancer for developing a cancer vaccine. ERas expression was determined in scirrhous gastric carcinoma specimens by immunohistochemical staining. To assess the possibility of the ERas protein as an anticancer vaccine target, we examined whether ERas for HLA-A-restricted epitope peptides were capable of eliciting cytotoxic T lymphocyte activity. Immunohistochemical analysis identified ERas protein in the nucleus and cytoplasm of cancer cells, yet ERas was not expressed in normal gastric epithelium. By western blotting, lysates of the scirrhous gastric cancer cell lines, OCUM-8, OCUM-2MD3 and OCUM-2M were shown to contain a 25-kDa band of ERas protein. ERas mRNA was detected in these cell lines by RT-PCR. To investigate cytotoxicity, we successfully established cytotoxic T lymphocyte clones stimulated by HLA-A*2402-restricted ERas peptides (FALDDPSSL). These peptides have specific cytotoxicity against corresponding HLA-A*2402-positive target cells pulsed with the candidate peptide. We found that the cytotoxic T lymphocyte clones demonstrated cytotoxic activity against OCUM-8 cells that endogenously express ERas. Our results suggest that ERas is a novel tumor-associated antigen with the potential application to be a vaccine against scirrhous gastric cancer.

Diagnostic significance of tumor markers.

Progress in the immunological detection of malignant tumors has been delayed by two major obstacles: (a) the lack of true cancer specificity of the known tumor-marker substances; and (b) the lack of a close relationship between the concentration and/or first appearance of the marker antigens and the malignant disease, especially at its early stage. An additional, important, and complicating factor implicating the specificity of the reactions of the test substances is the existence of immunological cross-reactions. Until recently, only two approaches have been pursued to resolve this difficulty of cross-reactivity, both concerned with the antigen. Now there is a third way, namely, the production of monoclonal antibodies. The poor correlation between the stage of the neoplastic disease and the level of circulating tumor markers, e.g., carcinoembryonic antigen, may be explained on a histological basis, or by inherent differences in the carcinoembryonic antigen molecule of different tumor sources. Few studies are available dealing with these problems.

Establishment of lymph node metastatic model for human gastric cancer in nude mice and analysis of factors associated with metastasis.

The actual mechanisms responsible for lymph node metastasis in gastric cancer are still unclear. To investigate the mechanisms of lymph node metastasis in gastric cancer, we established a lymph node metastatic model for human scirrhous gastric carcinoma. Lymph node metastasis had frequently developed after orthotopic implantation of OCUM-2M LN derived from a scirrhous gastric cancer cell line, OCUM-2M, which had low capacity for lymph node metastasis. We elucidated the different characteristics including binding ability, migratory capacity and immunoresponses induced by the cell surface molecules of these two cell lines. The binding ability to Matrigel and migratory capacity of OCUM-2M LN cells were significantly greater than those of OCUM-2M cells. On flow cytometric analysis, both OCUM-2M and OCUM-2M LN cells strongly expressed HLA-I (99.5 and 97.1%) and LFA-3 (76.6 and 99.2%) in level of expression between the two cell lines, but neither cell line expressed HLA-II (0.0 and 0.0%), B7-1 (0.0 and 0.0%) or B7-2 (0.4 and 0.3%). ICAM-1 expression in OCUM-2M LN cells was weaker (0.7%) than that in OCUM-2M cells (36.8%). Strong adhesiveness and cytotoxicity of mononuclear lymphocytes for OCUM-2M cells were observed in adhesion and cytotoxic assays, both of which were significantly decreased by the addition of anti-ICAM-1 antibodies. On the other hand, the adhesiveness and cytotoxicity of OCUM-2M LN cells were significantly less than those of OCUM-2M cells, and were not affected by the addition of anti-ICAM-1 antibodies. These findings suggest that decreased ICAM-1 expression in a new gastric cancer cell line with a high rate of lymph node metastasis may in turn decrease immune responses mediated through LFA-1-dependent effector cell adhesion, and that this escape from the immunosurveillance system may be one of the factors inducing lymph node metastasis. In conclusion, we established a gastric cancer cell line, OCUM-2M LN, with a high rate of lymph node metastasis. An in vivo lymph node-metastatic model with this cell line should be useful for analysing the mechanism and therapeutic approach of lymph node metastasis.

Stability and localization of biotin-labeled murine monoclonal antibody to human gastric cancer in normal mice and nude mice-human tumor models.

We examined the tissue localization of biotin-labeled murine monoclonal antibody (MAb) S202 directed against the human scirrhous gastric carcinoma cell line MK-01 in normal and tumor-bearing mice after intravenous (IV) administration. The biotin-labeled MAb proved to be stable in vivo under normal conditions, antibody titer being 1:256 at 4 hr after IV injection. At 24 hr after injection, the tumor was stained by the avidin-biotin-peroxidase complex (ABC) method. Biotin-labeled MAb was found to be suitable for detection of the xenografted tumor of nude mice. This study provides new information concerning the dynamics of the distribution of biotin-labeled MAb in vivo.

Regional hyperplastic lymph nodes in breast cancer: the role of lymphocytes and nodal macrophages. An immunological study with a five-year follow-up.

Five stage I and stage II breast cancer patients with sinus histiocytosis in two or more enlarged regional lymph nodes were studied. Peripheral lymphocytes, serum, and nodal lymphocytes were tested in vitro for cytotoxicity against autologous normal and tumor cells. Nodal macrophages were incubated with autologous peripheral lymphocytes and these "activated" lymphocytes then were tested in vitro for cytotoxicity against autologous normal and tumor cells. Peripheral lymphocytes (L) were not cytotoxic to autologous tumor (T) cells at 25:1 L/T ratios. Nodal lymphocytes were specifically cytotoxic to autologous tumor cells. Macrophages from hyperplastic regional lymph nodes transferred tumor specific inmunity to peripheral lymphocytes. Macrophages from small, nonhyperplastic regional lymph nodes did not transfer tumor specific immunity. With the advent of adjuvant chemotherapy and its attack on systemic immunity, a quantitative, immunopathological classification of breast cancer patients is needed in order to properly select patients for further therapy.

CEA positivity in sera and breast tumor tissues obtained from the same patients.

It is well known that the concentration of serum CEA correlates best with colorectal carcinomas but less well with other solid tumors. The aim of the present study was to elucidate further the poor relationship between confirmed primary mammary carcinomas and CEA serum levels, 108 of these tumors have hence been studied for their tissular CEA positivity and the results obtained were compared with the CEA serum values found preoperatively in the same patients. Techniques employed were the indirect immunoperoxydase method for the histologic studies and the enzymoimmuno-assay for the serum measurements. The discordance and concordance of the CEA results obtained with the two techniques were analyzed in respect to the histologic types of tumors classified following the ICD-O nomenclature. Our results seem to prove that: 1) although most tumors produce the antigen only a fraction of them releases it, and 2) the most common histologic type of tumor is the one secreting the antigen the less frequently. These observations are apparently independant of the tumor stages T1-T4, N- or N+, M-.

[Detection of antigens immunologically related to the envelope antigens of mouse mammary gland cancer virus antigens in human breast cancer tissue]. / Vyiavlenie v tkaniakh raka molochnoi zhelezy cheloveka antigenov, immunologicheski rodstvennykh obolochechnym antigenam virusa raka molochnoi zhelezy myshei.

A highly sensitive method of electroimmunodiffusion in acetate cellulose films developed by G.I. Abelev on the principle of isotachophoresis was used for the detection in human mammary gland carcinoma (HMGC) of an antigen related to envelope antigens of mouse mammary gland cancer (MMGC) virus. In 4 out of 10 HMGC cases, fractions of proteins with molecular weight about 50,000 d previously isolated in PAAG with sodium dodecyl sulphate were found to contain an antigen reacting with the antiserum to p52 MMGCV precipitation band which proved immunological identity of both antigens.

[Breast cancer associated with pregnancy].

Two cases of breast cancer associated with pregnancy were reported and the immunological assessment was performed in the second case. Case 1: A 36 year-old woman in the first trimester of pregnancy and having a large axillary mass and breast lump was admitted. She was diagnosed as having Stage IIIb breast cancer. After interruption of pregnancy, she received an extended radical mastectomy and radiotherapy. However, she died nine months after surgery for brain and liver metastases. Case 2: A 30 year-old woman in her second trimester of pregnancy was admitted for a large breast mass. She was diagnosed as having Stage IIIa breast cancer. Interruption of pregnancy was again necessary to perform surgery followed by chemotherapy. After an extended radical mastectomy she was placed on a combination chemotherapy regimen with cyclophosphamide, adriamycin and 5-FU. In both cases tumors began to develop rapidly at pregnancy. Immunological studies in Case 2 showed a depression of T-lymphocyte function and NK cell activity. Our study suggests that the depression of cell-mediated immunity during pregnancy may promote tumor growth.

Cellular and stromal characteristics in the scirrhous hepatocellular carcinoma: comparison with hepatocellular carcinomas and intrahepatic cholangiocarcinomas.

Scirrhous hepatocellular carcinoma (SHCC) is a rare variation of HCC, for which characteristics of tumor cells and the fibrotic stroma have not been clarified in detail. The present study was therefore carried out to elucidate cytological features of tumor and stromal cells and components of the stromal extracellular matrix in 15 SHCC patients undergoing hepatectomy without preoperative transarterial embolization. Diagnosis was on the basis of a scirrhous histological pattern exceeding 50% of the tumor area. Expression of cytoplasmic and extracellular matrix proteins was compared among SHCC, HCC and intrahepatic cholangiocarcinoma (ICC) cases with immunohistochemical staining. The lesions could be histologically divided into radiating and sinusoidal types. Common stromal components of SHCC and ICC were collagen types I and III. There was no expression of laminin-5 in the stroma of SHCC, but it was present in almost all ICC cases. Tenascin-C expression was significantly lower in the SHCC cases and its distribution differed between SHCC and ICC. Matrix metalloproteinase-7 (MMP-7) expression was significantly higher in SHCC compared with HCC. Almost all stromal cells were alpha-smooth muscle actin-positive both in SHCC and ICC, whereas glial fibrillary acid protein (GFAP)-positive stromal cells were significantly more increased in ICC than in SHCC. SHCC clearly differed from HCC with respect to collagen types I, III and MMP-7 expression, and from ICC with regard to stromal components including laminin-5, tenascin-C and GFAP(+) stromal cells.

Monoclonal antibodies against human scirrhous carcinoma of the stomach.

We established three monoclonal antibodies (MoAbs) S43, S74 and S202 which reacted strongly with scirrhous carcinomas of the stomach. MoAb S43 reacted with culture supernatants of the scirrhous gastric cancer cell line MK-01, and thus may be useful for in vivo detection of markers for this tumor. MoAb S74 reacted with the majority of gastric cancer tissues, but showed no reaction with normal tissues. It may be useful as a drug carrier in targeting chemotherapy against scirrhous gastric cancer, because of its tumor specificity. MoAb S202 reacted strongly with 100% of scirrhous gastric cancer samples and with the culture supernatants of colonic and pancreatic cancers.

Localization of radiolabeled monoclonal antibody S74 in human scirrhous type gastric cancer xenografts in nude mice.

A mouse monoclonal antibody (MoAb), S74 against a human scirrhous gastric carcinoma cell line was developed. The radioiodinated antibody bound preferentially in vivo to xenografted tumors rather than to normal tissue, whereas tumor uptake of normal mouse IgG did not occur. Thus this MoAb was able to selectively localize a human tumor in an animal model, and clinically it may enable immunotherapy against scirrhous carcinoma of the stomach.

[Immunomodulation by leukocytolysis. On the significance of lectin-polysaccharide complexes]. / Immunmodulation durch Leukozytolyse. Zur Bedeutung von Lektin-Polysaccharid-Komplexen.

Polymorphonuclear granulocytes (PMNs) are thought to function only as phagocytes. The ability of PMNs for a physiologically lysis has not been noticed so far. Therefore, PMNs are not only a clamp between the specific and unspecific immune system but an important regulator of the homoeostasis. All immune modifiers therapeutically used seem to stimulate the physiologically lysis of PMNs. This has been shown with mistletoe lectins. Most remarkable are findings that show lectins bound to polysaccharides drastically rise the ability of PMNs to undergo lysis.

A monoclonal antibody reactive with gastric carcinoma.

A monoclonal antibody reactive with gastric carcinomas of the stomach was established. This antibody reacted with 88% (15/17) of the scirrhous carcinomas, and 75% (6/8) of the well differentiated adenocarcinomas of the stomach tested, but did not react with normal gastric mucosa or various other normal tissues, with the exception of the mucosa of the large intestine.

[Carcinoembryonic antigen in serum and tissue of benign and cancerous breast lesions]. / L'antigène carcino-embryonnaire (ACE) Sérique et tissulaire dans les lésions bénignes et canceréuses du sein.

The tissular CEA was studied with the immunoperoxidase method and the circulating CEA with the enzyme immunoassay (EIA), using peroxidase-coupled antibodies. 51 cases of mammary cancers as well as 33 benign lesions were examined in order to study a possible correlation between the frequency of CEA-positive tissues and CEA-positive sera in the same patient. It has been confirmed that CEA-positive tumors can be accompanied by low or negative CEA serum levels. An attempt was made to analyze the stage and the histologic type of lesions in comparison to the positivity and the increase of CEA.

A case report of immunotherapy on a patient with advanced gastric cancer by adoptive transfer of OK-432-reactive HLA-matched allogeneic lymphocytes.

Adoptive immunotherapy (AIT) for non-hematological malignancies, using HLA-matched donor lymphocytes, has been rarely reported. For a 35-year-old male patient with peritoneal disseminated advanced gastric cancer, we performed AIT using lymphocytes from his HLA-matched 37-year-old brother and a streptococcal preparation, OK-432, as an antigen. After the donor had been immunized by intradermal administration of OK-432, OK-432-reactive lymphocytes were induced in vitro and transferred to the patient intravenously with OK-432. Low-dose systemic immunochemotherapy, using interleukin-2, 5-fluorouracil and cyclophosphamide, was concurrently administered with AIT. As a result, the Schnitzler metastasis in the patient reduced in size without any significant graft-versus-host-related complications. One of the effector mechanisms of therapeutic benefit was suggested to be cytokine release from the transferred OK-432-reactive lymphocytes. Our findings suggest the safety and efficacy of AIT using lymphocytes from an HLA-matched sibling and OK-432 as an antigen. Further studies to investigate the use of tumor-associated antigen and an HLA-matched sibling's lymphocytes for AIT of advanced cancer are warranted.

[A monoclonal antibody reactive with scirrhous carcinoma of the stomach].

A monoclonal antibody reactive with human gastric carcinoma was established. When formalin-fixed, paraffin-embedded histological sections were stained by avidin-biotin-peroxidase complex method using this monoclonal antibody, it was found to react with 89% (17/19) of the scirrhous carcinomas (diffuse type of carcinoma) and 72% (13/18) of the differentiated adenocarcinomas of the stomach tested. However, this antibody did not react with normal gastric mucosa or various other normal tissues except the mucosa of the large intestine.

Localization of antigen defined by anti-scirrhous gastric carcinoma monoclonal antibody S202 in fixed human cancer tissues.

Monoclonal antibody S202 reactive with scirrhous gastric carcinoma cell line MK-01 was used in an immunoperoxidase assay to characterize the antigenic profile of a variety of fixed samples. Representative sections of malignant tissue from the stomach, colon, esophagus, pancreas, bile duct, gallbladder and breast were examined by the avidin-biotin complex immunoperoxidase technique and compared. The antibody reacted with 90 out of 98 stomach adenocarcinomas, and with 30 out of 32 colon adenocarcinomas with diffuse cytoplasmic staining, whereas it reacted with only 19 out of 37 breast carcinomas, and 5 out of 10 esophageal squamous cell carcinomas. The antigenic determinants appeared to be a carbohydrate, since it was highly sensitive to sodium periodate.

A tumor-associated antigen in the scirrhous gastric carcinoma cell line MK-01 defined by monoclonal antibody S202.

A monoclonal antibody (MAb) S202, with an IgG1 isotype, that reacted strongly with the scirrhous gastric carcinoma cell line MK-01 was established. MAb S202 reacted with the colonic cancer cell line, SW1116, and the pancreatic cancer cell line, PK-1, when tested by indirect immunofluorescence. The S202 reactive antigen was expressed in the majority of acetone-fixed fresh frozen cancer tissues. Eighty to 100 per cent of the paraffin-embedded sections of stomach, colon and pancreatic adenocarcinoma were positive for the S202 antigen, with diffuse cytoplasmic staining, whereas esophageal and breast cancers demonstrated markedly less immunostaining. Supplemented serum-free medium collected from 7 day old tumor cell cultures were assayed for the presence of antibody-defined antigens. Antigens detected by MAb S202 were released by the cell lines SW1116 and PK-1. The binding of MAb S202 to the colonic adenocarcinoma sections was reduced after treatment with sodium periodate which suggests that respective antigenic determinants are of carbohydrate nature without sialic acid residues.