Fucosylation of serum alpha-fetoprotein in patients with primary hepatocellular carcinoma.

alpha-Fetoprotein specimens were prepared from the sera of four patients with hepatocellular carcinoma. The lentil lectin-reactive and lectin-nonreactive variants of this glycoprotein were also prepared from the serum of one of the four patients by affinity chromatography with immobilized lectin. The correlation between the carbohydrate structure of these compounds and their reactivity in crossed immuno-affinoelectrophoresis with lentil lectin was studied by chemical analysis and affinity chromatography of the glycopeptides with lectin columns. It was found that the lentil lectin-reactive variant contained a carbohydrate chain of the fucosylated biantennary complex type. These data together with previous findings indicate that most of the patients with hepatocellular carcinoma have an elevated serum concentration of fucosylated alpha-fetoprotein.

Improvement in gastric histology following Helicobacter pylori eradication therapy in Japanese peptic ulcer patients.

We aimed to determine if successful or failed eradication of Helicobacter pylori with triple therapy causes any difference in gastric mucosal histology. Japanese H. pylori-positive patients with a healed peptic ulcer received high (n = 112) or low (n = 113) doses of triple therapy (omeprazole, amoxicillin and clarithromycin) for 1 week. Biopsies from the greater curvature of the central antrum and upper corpus were taken 6 weeks and 30 weeks after treatment completion, and gastric mucosal histology compared between successful (n = 171) and failed (n = 34) eradication groups. Morphological variables of gastritis were graded according to the updated Sydney System. Successful eradication therapy was defined as improvement in inflammation, neutrophil activity and atrophy; failed eradication therapy as improvement in inflammation and neutrophil activity only. Gastric mucosal atrophy gradually improved (in addition to improvements in inflammation and neutrophil activity) with successful eradication of H. pylori infection.

[Usefulness of contrast-enhanced CT of the peritoneum to evaluate efficacy of neoadjuvant chemotherapy for peritoneal dissemination in an advanced gastric cancer case].

We performed contrast-enhanced CT before and after chemotherapy in a patient with advanced gastric cancer to assess the efficacy of neoadjuvant chemotherapy for peritoneal dissemination. The patient was preoperatively given combined chemotherapy with UFT and CDDP. Hepatic metastatic and peritoneal disseminated foci were markedly reduced on CT. Thus CT proved to be useful for assessing peritoneal dissemination and the efficacy of neoadjuvant chemotherapy.

[Effect of neoadjuvant chemotherapy (CDDP.UFT therapy) upon cell DNA analysis by flow cytometry in gastric carcinoma].

Neoadjuvant chemotherapy with CDDP and UFT was performed on far advanced gastric carcinoma cases whose curative resection was impossible. These trials were carried out on 10 patients who had Borrmann 4 type carcinoma. PR was found in 6 cases, for an efficacy rate of 60% (6/10). The DNA content in gastric carcinoma was determined before and after treatment by flow cytometry. The relationship between the change of DNA ploidy pattern and effects of chemotherapy was investigated. Aneuploidy was found in 6 of 10 patients (60.0%). DNA ploidy patterns showed no correlation with effectiveness. In aneuploid cases, the group of responders showed seemingly the change of ploidy pattern, from aneuploid to diploid. In non-responders the percentage of aneuploid cells increased. In diploid cases, an accumulation of S-phase fraction was observed in the group of responders. These results suggested that DNA ploidy analysis may provide additional diagnostic criteria for better assessment of chemotherapeutic effects.

The reactivity of alpha-1-antitrypsin with Lens culinaris agglutinin and its usefulness in the diagnosis of neoplastic diseases of the liver.

The reactivity of alpha-1-antitrypsin (AAT) with Lens culinaris agglutinin (LCA) was studied by crossed immuno-affinity electrophoresis of the sera of 246 subjects from 6 groups (acute virus hepatitis, chronic hepatitis, liver cirrhosis, hepatocellular carcinoma (HCC), carcinoma metastatic to the liver and normal controls). Two species of AAT (LCA-reactive and -nonreactive species) were detected on crossed immuno-affinity electrophoresis in a gel containing LCA. The percentages of LCA-reactive species of AAT in neoplastic diseases of the liver were significantly higher than those in benign liver diseases and normal controls. There was no correlation between the percentage of LCA-reactive species of AAT and serum AAT concentration in any group. Furthermore, in studying 15 pairs of serum samples before and after the subsequent development of HCC, the percentage of LCA-reactive species of AAT after HCC occurrence was significantly higher than that before, although there was no statistically significant difference between the serum AAT concentration before and after development of the disease. The latter 15 patients were all of the normal protease inhibitor phenotype (PiMM) and no change in phenotype was observed before and after the development of HCC. The results indicate that measurement of the reactivity of AAT with LCA can be a useful marker for the diagnosis of HCC and carcinoma metastatic to the liver, especially when serum concentrations of alpha-foetoprotein or other tumour markers are within the normal ranges.

Expression and function of CD40 in rheumatoid arthritis synovium.

OBJECTIVE: To determine the involvement of CD40 in chronic activation of rheumatoid arthritis (RA) synovial monocytes. METHODS: CD40 expression on RA synovial monocytes was examined by immunostaining. Involvement of CD40 in tumor necrosis factor-alpha (TNF-alpha) secretion from RA synovial fluid mononuclear cells (SFMC) was examined by blocking with anti-CD40 antibody. RESULTS: CD40 was expressed on RA synovial monocytes. TNF-alpha secretion from RA SFMC was enhanced by CD40 ligand stimulation. Spontaneous secretion of TNF-alpha from RA SFMC was inhibited by anti-CD40 antibody. CONCLUSION: CD40 was involved in the activation of RA synovial monocytes that leads to TNF-alpha production.

Roles for integrin very late activation antigen-4 in stroma-dependent erythropoiesis.

Adhesion molecules are required for development of hematopoietic stem and progenitor cells in the respective hematopoietic microenvironments. We previously showed that development of the erythroid progenitor cells is dependent on their direct adhesion to the stroma cells established from the erythropoietic organs. In this stroma-dependent erythropoiesis, we examined the role of adhesion molecules in erythropoiesis by blocking antibodies. The development of the erythroid cells on stroma cells was inhibited by anti-very late activation antigen-4 (VLA-4 integrin) antibody, but not by anti-VLA-5 antibody, although the erythroid cells express both VLA-4 and VLA-5. Whereas high levels of expression of vascular cell adhesion molecule-1 (VCAM-1) and fibronectin, ligands for VLA-4, were detected in the stroma cells, the adhesion and development of the erythroid progenitor cells were partly inhibited by the blocking antibody against VCAM-1. VLA-5 and fibronectin could mediate adhesion of the erythroid progenitor cells to the stromal cells, but the adhesion itself may not be sufficient for the stroma-supported erythropoiesis. The stromal cells may support erythroid development by the adhesion through a new ligand molecule(s) for VLA-4 in addition to VCAM-1, and such collaborative interaction may provide adequate signaling for the erythroid progenitor cells in the erythropoietic microenvironment.

The fucosylation index of alpha-fetoprotein and its usefulness in the early diagnosis of hepatocellular carcinoma.

The serum concentration and degree of fucosylation (fucosylation index) of alpha-fetoprotein (AFP) were determined in serum samples from 258 patients with hepatocellular carcinoma (HCC) and 114 patients with benign liver diseases. When the serum AFP concentration was below 1000 ng/ml, it could not be used as a measure to distinguish between HCC and benign liver diseases. However, the fucosylation index of AFP proved useful for such a purpose. The sensitivity of the analysis using the fucosylation index in total patients with HCC was 69%; the specificity was 96% in benign liver diseases, and the accuracy of this test was 77%. When HCC patients who were grouped according to tumor size (5 cm, 3 cm, and 2 cm in diameter) were analyzed, they all had a fucosylation index significantly higher than that in benign liver disease patients. The mean fucosylation index in 28 patients with a serum AFP concentration below 1000 ng/ml and a tumor diameter less than 3 cm was 26 +/- 30%. Corresponding values for 16 patients with an AFP concentration below 400 ng/ml and a tumor size less than 3 cm and for 8 patients with a concentration below 400 ng/ml and a tumor size less than 2 cm were 32 +/- 31% and 27 +/- 27%, respectively. These values were higher, with statistical significance, than those in patients with benign liver diseases. These data indicate that the measurement of the fucosylation index of AFP is useful for the early diagnosis of HCC.

Close topographical relationship in alpha foetoprotein (AFP) between a lens culinaris binding glycan and the epitope recognized by AFP-reactive monoclonal antibody, 18H4.

Monoclonal antibodies 18H4 and 19F12 against alpha-foetoprotein (AFP) were examined by enzyme immunoassay for binding to two forms of AFP that were separated on the basis of the reactivity with lentil lectin (LCA). LCA-binding and LCA non-binding AFP, coated on a solid phase, reacted with 18H4 but reactivity with the LCA-binding species was inhibited by 60% following pretreatment of the AFP with LCA. The lectin was a very poor inhibitor of binding of 18H4 to the AFP which did not interact with LCA. In an alternative binding assay, a polyclonal anti-AFP coated solid phase was reacted with beta-galactosidase-labelled 18H4. Pre-treatment with LCA of the LCA-reactive AFP gave 56% inhibition of binding of conjugated 18H4 while little inhibition was achieved with the LCA-nonreactive AFP component. These findings show that the epitope recognised by 18H4 is distinct from the glycan sequence that is reactive with LCA. However, the LCA-binding oligosaccharides occur in close proximity to the 18H4 epitope in the native AFP.

Fas-mediated stimulation induces IL-8 secretion by rheumatoid arthritis synoviocytes independently of CPP32-mediated apoptosis.

In this study, we investigated the IL-1 beta converting enzyme (ICE) family cysteine proteases responsible for the Fas-mediated apoptosis of rheumatoid arthritis (RA) synoviocytes and their involvement in proinflammatory cytokine production. CPP32 inhibitor, but not ICE inhibitor, was capable of inhibiting the Fas-mediated apoptosis of RA synovial cells. CPP32, but not ICE, was activated in response to anti-Fas stimulation. IL-8, but not IL-1 beta, was secreted from the anti-Fas-stimulated RA synoviocytes even in the presence of CPP32 inhibitor. These results demonstrated that CPP32, but not ICE, is the predominant cysteine protease that mediates the Fas-mediated apoptosis of RA synovial cells. We also demonstrated that anti-Fas stimulation of RA synoviocytes leads to IL-8 secretion independently of the CPP32-mediated apoptosis, which would accelerate inflammation.

Expression and function of fibronectin binding integrins on rat mast cells.

Adhesion molecules of the integrin family are implicated not only in leukocyte migration but also in leukocyte activation. Here we characterize the expression and function of fibronectin receptor integrins on rat mast cells. A rat basophilic leukemia cell line (RBL-2H3) and phorbol ester-stimulated rat peritoneal mast cells adhered to fibronectin (FN), vitronectin and fibrinogen. These mast cells expressed fibronectin receptor integrins, including very late antigen (VLA)-4, VLA-5 and vitronectin receptor (VNR), as estimated by immunofluorescent staining and inhibition of FN adherence by newly established mAbs reactive with the rat alpha 4 (MR alpha 4-1), alpha 5 (HM alpha 5-1) or beta 3 (HM beta 3-1) chains of the integrin molecules. The beta-hexosaminidase release, a marker for mast cell degranulation, triggered by high affinity IgE receptor (Fc epsilon RI)-mediated stimulation, was enhanced by adhesion of RBL-2H3 cells to either immobilized FN, MR alpha 4-1, HM alpha 5-1 or HM beta 3-1. This FN enhancement of beta-hexosaminidase release was inhibited by soluble MR alpha 4-1, HM alpha 5-1 and HM beta 3-1 as well as by GRGDSP and DELPQLVTLPHPNHLGPEILDVPST peptides which abrogate VLA-5/VNR and VLA-4 binding to FN respectively. In vivo, passive cutaneous anaphylaxis induced by IgE anti-DNP and DNP-BSA was inhibited by concurrent s.c. injection of MR alpha 4-1, HM alpha 5-1 and HM beta 3-1. These results demonstrate that FN receptor integrins expressed on rat mast cells play an important role in regulating mast cell activation both in vitro and in vivo.