Hideyuki Yoshioka

Differential expression of human spasmolytic polypeptide (trefoil factor family-2) in pancreatic carcinomas, ampullary carcinomas, and mucin-producing tumors of the pancreas

Human spasmolytic polypeptide (hSP) is a member of the trefoil peptide group, thought to be involved in mucin production and cell growth. It has been reported that hSP protein is expressed in digestive cancers but not in normal pancreas. The expression of hSP in pancreatic neoplasms has not been investigated in detail. The immunohistochemical expression of hSP protein was investigated in pancreatic carcinomas, ampullary carcinomas, mucin-producing tumors, serous cystadenomas and islet cell tumors of the pancreas. hSP was expressed in 23% of pancreatic duct cell carcinomas, and hSP protein was more frequently detected in cases of early-stage or histologically low-grade duct cell carcinomas than in cases of late-stage or histologically high-grade carcinomas. Patients with hSP protein expression showed a better prognosis than did those with negative hSP expression. hSP expression was detected in 92% of mucin-producing tumors, but was not detected in serous cystadenoma or islet cell tumors. Immunohistochemical hSP expression is related to differentiation and a better prognosis in pancreatic duct cell carcinomas. Furthermore, hSP protein is related to the pathogenesis and clinical characteristics of mucin-producing tumors of the pancreas.

Circulating immune complex, endotoxin, and biliary infection in patients with biliary obstruction

Immunoglobulin A-containing circulating immune complexes, immunoglobulin G-containing circulating immune complexes, and endotoxin were measured in the sera of patients with obstructive jaundice. The bile of patients with percutaneous transhepatic biliary drainage was also cultured for bacteriologic studies. There was a significantly positive correlation between the endotoxin levels and both immunoglobulin A-containing circulating immune complex and immunoglobulin G-containing circulating immune complex. The endotoxin levels of the patients with gram-negative infections were significantly increased compared with those of the patients with sterile cultures. The immunoglobulin G-containing circulating immune complex levels of the patients with bacteria in bile were significantly increased compared with those of the patients with sterile cultures. The immunoglobulin A-containing circulating immune complex levels of the patients with bacteria in bile were slightly increased, but the difference did not reach statistical significance. These results indicate that one of the causes of increased circulating immune complex levels may be endotoxemia in combination with biliary infection in patients with biliary obstruction.

Expression of sialosyl-Tn antigen (monoclonal antibody MLS102 reactive) in normal tissues and malignant tumors of the digestive tract

Oncogenic transformation is often associated with changes in the glycosylation state of malignant cells. We investigated the immunohistochemical localization of sialosyl-Tn antigen [O-linked NeuAc(α2→6)GAINAc] using a novel monoclonal antibody MLS102 in normal and malignant digestive-tract tissues. In normal tissues, weak MLS102 immunoreactivity was observed in the epithelium of the esophagus, stomach and colon. However, MLS102 immunoreactivity was strong in the goblet cells of the duodenum, but not in the Brunner glands. In carcinomas of the esophagus, stomach, colon, pancreas and biliary tract, positive staining was detected with a high frequency (80%–100%). In mucinous carcinomas and signet-ring cell carcinomas, malignant cells themselves and the mucins they secreted were strongly positive for sialosyl-Tn antigen. There was no significant correlation between the frequency of expression of sialosyl-Tn antigen and the degree of differentiation (grade). However, in the case of well-differentiated adenocarcinomas, sialosyl-Tn antigen was found mainly in the supranuclear areas (Golgi area), on the apical surface and in the adjacent cytoplasm. In poorly differentiated adenocarcinomas, the antigen was often detected in the whole plasma membrane and cytoplasm. Therefore, monoclonal antibody MLS102 may be useful in further elucidating the characteristics of digestivetract cancers, and possibly in their treatment.

High Levels of Anti-Mitochondrial Antibodies in MRL Mice Influence of Anti-DNA Antibodies on Anti-Mitochondrial Antibodies Measured by an Enzyme-Linked Immunosorbent Assay

We developed an easy enzyme‐linked immunosorbent assay (ELISA) for anti‐mitochondrial antibodies, and detected high levels of anti‐mitochondrial antibodies in MRL/Mp‐lpr/lpr (MRL 1/1) mice. The influence of the presence of anti‐DNA antibodies in the tested sera on this assay was also evaluated. The monoclonal anti‐DNA antibodies, which were made from non‐immunized MRL 1/1 mice, did not react with the mitochondrial antigens adhering to polystyrene plates. Absorption with DNA had little effect on the levels of mitochondrial antibodies in MRL 1/1 sera. Our assay for mitochondria‐binding antibody was very easy and sensitive, although it could not differentiate among heterogeneous anti‐mitochondrial antibodies.

High Levels of IgA-Containing Circulating Immune Complex and Secretory IgA in Kawasaki Disease

Sera of patients with Kawasaki disease were studied for the levels of IgA‐containing (C3‐fixing) circulating immune complexes (IgA‐CIC), IgG‐containing (IgG‐) CIC, total IgA, secretory IgA, and complement component (C3) by means of enzyme‐linked immunosorbent assays or single radial immunodiffusion methods. There was significantly high level of IgA‐CIC, but not IgG‐CIC. The levels of total IgA, secretory IgA, and C3 were significantly elevated. Significantly high levels of secretory IgA were found in 22 (51%) of 43 patients. The proportion of secretory IgA to total IgA also increased. These abnormalities in the IgA system may play a role in Kawasaki disease.

Immunohistochemical examination of Peyer's patches in senescence-accelerated mice.

The distribution of T cells, B cells and murine senile amyloid protein in Peyers Patches was examined in senescence accelerated (SAM-P/1) and control (SAM-R/1) mice ranging in age from two to ten months. An immunohistochemical detection of lymphocyte surface antigens of T cells and B cells. A murine senile amyloid protein in Peyers patches was detected by immunohistochemical staining with the specific antiserum. Congo red staining and electron microscopy. The T cell population increased and B cell population decreased slightly with age in SAM-P/1 mice, but little change was observed with age in SAM-R/1 mice. Murine senile amyloid protein positive spots were seen surrounding the vessels of the thymus-dependent area at 6 months of age, and were observed throughout the Peyers patches at 10 months of age, but were not observed by 10 months of age in SAM-R/1 mice.

Effects of percutaneous transhepatic biliary drainage on blood-bile permeability and selective IgA transport in patients with biliary obstructions.

Biliary obstruction induces a increase in the permeability between blood and bile, and a decrease in the rate of immunoglobulin A (IgA) transport into bile. We investigated the effects of percutaneous transhepatic biliary drainage (PTBD) on these derangements. PTBD reduced the extent of elevation of the bile-to-serum ratio of Immunoglobulin G (IgG; IgG-BS ratio) in patients with obstructive jaundice. Because IgG is known to be passively transported from serum to bile, the results indicate that PTBD restores the blood-bile barrier function. The IgA-BS ratio/IgG-BS ratio index (IgA/IgG index) and the IgM/IgG index, which indicated the function of selective transport of IgA and IgM into bile, initially decreased and then returned to the normal range 17 days after PTBD in patients who experienced a rapid resolution of hyperbilirubinemia. However these indices remained low in patients who did not experience this resolution. The serum secretory IgA levels in patients who did not experienced rapid resolution of hyperbilirubinemia markedly increased before PTBD. The serum secretory IgA levels in the patients who did and those who did not experience rapid resolution of hyperbilirubinemia, after initially increasing, decreased after PTBD. However the level returned to the control range only in patients who experienced a rapid resolution. These results indicate that the secretory IgA level is a sensitive indicator of hepatobiliary function, and measurement of the level of secretory IgA could predict the effect of PTBD.

Immunohistochemical examination of Peyer's patches in autoimmune mice

SummaryThe distribution of T-cells and B-cells in Peyers patches was examined in three autoimmune model mice, MRL/Mp-lpr/lpr, BXSB, NZBWF1/J mice and normal BALB/c mice, between one and ten months old. A multiple layering technique was used for immunohistochemical detection of lymphocyte surface antigens of T-cells (Thy1.2, Lyt1, Lyt2) and B-cells (surface IgM) and peanut agglutinin receptor for germinal center cells. The T-cell population of female MRL/Mp-lpr/lpr mice increased markedly with age, and the B-cell population of the male BXSB mouse tended to increase. However, little change was observed with age in the NZBWF1/J mice. The immunohistochemical properties of the Peyers patches in the three autoimmune model mice were different.

Age Associated Changes in the Distribution of Ipr Gene-Induced B220-Positive T Cells in Lymphoid Organs of MRL/Mp-Ipr/Ipr Mice Using Dual Exposure Microphotographs of Double Immunofluorescence Staining

Homozygous MRL/Mp-lpr/lpr [MRL/lpr] mice, which have an autosomal recessive mutant lpr gene and exhibit defects in Fas antigen, spontaneously develop autoimmune disease with progressive expansion and accumulation of characteristic abnormal CD4-CD8-double negative T cells that express B220 surface antigen, a B cell-specific surface marker in normal mice. We analyzed the distribution and age related changes of lpr gene-induced abnormal T cells (B220-positive lpr T cells) in the lymphoid organs of MRL/lpr mice. We studied cryostat sections of the spleen, peripheral lymph nodes, mesenteric lymph nodes, and Peyers patches at different stages using FITC [fluorescein isothiocyanate)-conjugated monoclonal antibodies directed against B220 (RA3-6B2) and PE (phycoerythrin)-conjugated anti-mouse CD3 (2C11) monoclonal antibody, examining dual-exposure microphotographs of double-immunofluorescence stained preparations. We observed that in aged MRL/lpr mice, B220-positive abnormal lpr T cells were not present in the thymus-dependent area, and the majority of the follicular area cells were displaced by lpr T cells. These findings suggest that the cellular trafficking of B220-positive lpr T cells differs from that of conventional T cells and that these lpr-derived T cells play a role in the follicle.

Immunohistochemical demonstration of a new thiamine diphosphate-binding protein in the rat digestive tract

SummaryWe purified a new thiamine diphosphate-binding protein (ThDP-BP), produced an antiserum against it, and examined its immunohistochemical distribution in the rat gastrointestinal tract using the avidinbiotin complex method. Positive staining for ThDP-BP was found in the epithelial glands of the stomach, small intestine and large intestine, and in the nuclei of hepatocytes. Measurement of the content of thiamine and its phosphate esters in extracts from the gastric and intestinal mucosa also indicated the presence of ThDP-BP in the stomach and intestinal mucosa. ThDP-BP may be useful for investigating the absorption and metabolism of the thiamine in metabolic disorders.