Activation of mRNA expression of collagen,collagenase and its inhibitor on renal biopsy specimens in patients with IgA nephropathy

Summary: In situ hybridization of mRNA for collagen IV, collagen VI, stromelysin (MMP-3) and TIMP1 was examined in renal biopsy specimens from patients with IgA nephropathy (IgAN) or diabetic nephropathy with various degrees of tissue damage. The majority of cells in the glomeruli expressed these mRNA almost simultaneously, but a few cells demonstrated positive expression for only one of these probes. There was a parallel relationship between the degree of tissue damage and that of mRNA expressions of these probes in patients with IgAN, while patients with diabetic nephropathy showed a reverse relationship between these two parameters. It is concluded that patients with mesangial proliferative glomerulonephritis expressed mRNA for collagen collagenase and its inhibitor in the glomeruli in parallel with the progress of tissue damage. In contrast, glomerular samples from patients with diabetic nephropathy showed that there was an inverse relationship between tissue damage and expression of mRNA. It is concluded that expression of collagen, collagenase and its inhibitor parallels the progression of glomerular changes in IgAN, but such parallel expression was not observed in patients with diabetic nephropathy.     

Effects of hydrocortisone and adrenaline on natural killer cell activity

We have studied the effects of hydrocortisone and adrenalineon natural killer (NK) cell activity and on the distributionof circulating lymphocyte sub-populations in 30 patients undergoingelective partial laminectomy under general anaesthesia. Thepatients were allocated to receive adrenaline (group 1, n =11), hydrocortisone and adrenaline (group 2, n = 11) or neitherhydrocortisone nor adrenaline (group 3, n = 8). Group 1 andgroup 2 patients received local adrenaline infiltration duringoperation to reduce bleeding. The mean dose of adrenaline administeredwas 2.1 (SD 0.2) µg kg^–1 Group 2 received hydrocortisone10 mg kg^–1 i.v. after premedication. In groups 1 and 2,adrenaline produced an instantaneous increase in NK cell activityaccompanied by a selective increase in circulating NK cells.The measurements returned to pre-infiltration levels within120 min of administration of adrenaline. The effect of adrenalinein causing increased NK cell activity was not blocked by pre-administrationof hydrocortisone. There was a significant decrease in the ratioof T helper/inducer cells (CD4) to T-suppressor/ cytotoxic cells(CD8) in all patients after induction of anaesthesia. In groups1 and 3, the CD4/CD8 cell ratio did not change significantlyduring operation. However, compared with groups 1 and 3, group2 showed a significantly reduced CD4/CD8 cell ratio during operation.Therefore, these results suggest that even in cases of suchsevere stress that the immune response was depressed by increasedserum cortisol concentrations, adrenaline-induced NK cell activityenhancement was preserved.     

Specific immunotherapy with tumour-draining lymph node cells cultured with both anti-CD3 and anti-CD28 monoclonal antibodies

For providing costimulatory signals, we utilized anti-CD28 monoclonal antibody (mAb) for the in vitro culture of tumour-draining lymph node (LN) cells. The proliferation of B16 melanoma-draining LN cells in the culture with anti-CD3 mAb was remarkably enhanced by the addition of anti-CD28 mAb. In culture with both anti-CD3 and anti-CD28 mAb, the B16-draining LN cells produced a higher level of interferon-γ, but not interleukin-4, than with anti-CD3 mAb alone. The B16-draining LN cells efficiently expanded in the culture with both anti-CD3 and anti-CD28 mAb and subsequently with a low dose of IL-2 (anti-CD3 plus anti-CD28/IL-2). The expanded cells consisted predominantly of CD8⁺ T cells and showed a specific cytolytic activity, in a major histocompatibility complex (MHC) class I-restricted manner, even without in vitro restimulation. In addition, the adoptive transfer of the B16-draining LN cells, expanded in the culture protocol of anti-CD3 plus anti-CD28/IL-2, showed a significant anti-tumour effect against metastatic B16 melanoma in combination with IL-2. The cured mice thus acquired a specific protective immunity. Moreover, this protocol was also moderately effective against poorly immunogenic 3LL carcinoma. Overall, our results suggest the potential for another immunotherapeutic strategy based on ‘the costimulatory theory’ other than vaccination with B7-transfected tumour cells.     

A Study on HL—A System in Japanese

Two hundred unrelated Japanese individuals were HL-A typed with UCLA Research Tray T3 (Terasaki's Tray), which contains specificities added after the Fifth International Workshop. Phenotype, gene and haplotype frequencies were calculated with standard errors and delta values. HL-A9, HL-A5 and W10 had a higher frequency and HL-A1, 3 and 8 had a lower frequency in Japanese than in Caucasians. The frequent haplotypes were HL-A9-HL-A5, HL-A9-HL-A7 and HL-A2W10. HL-A9-HL-A5 showed very positive high linkage disequilibrium parameter (delta value) and HL-A9-W10 showed negative high value. The sera designated as anti-HL-A, W5 and W15 in the T3 Tray which react identically in Caucasians showed different patterns of reaction when tested in the Japanese population. Five hundred Japanese parous women's sera were tested for cytotoxic antibodies. Some Japanese antisera showed high correlation coefficient values on HL-A2, HL-A9, HL-A10, HL-A11 and HL-A12. The women providing the anti-HL-5 complex sera and their immunizing persons were HL-A typed. These complex sera reactions were compared with the antisera in the T3 Tray. A new group of sera (SN-1), "operationally monospecific" and cross-reacting with W22, was found in the present study. Population and family studies suggested that the sera SN-1 are third in frequency within the second series (phenotypic frequency 17-22%) and show high delta values with HL-A11 in the Japanese population.     

Early sigmoid colon carcinoma developed from a pedunculated polyp

A 60‐year‐old man underwent routine colonoscopy, and was noted to have a pedunculated polyp in the sigmoid colon. The pathologic diagnosis was adenoma, and due to patient’s personal circumstances, the lesion was left untreated. The colonoscopic examination was repeated 4 years and 11 months later, to find that the polyp had transformed into an elevated lesion with irregularly depressed surface. The patient was diagnosed as having early sigmoid cancer, and underwent sigmoidectomy. The histologic examination of the excised specimen revealed well‐differentiated adenocarcinoma with invasion into the submucosal layer. Through studying the natural course of colon cancer, it has become known that the advanced cancers commonly develop from polyps with short pedicles (sessile polyps). This case represents an early sigmoid cancer developed from a pedunculated polyp, which differs from the current mainstream concept of ‘polyp‐cancer sequence of colon cancer.’     

The Role of Superoxide Anion and Lysosomal Enzymes in Anti-Listerial Activity of Elicited Peritoneal Macrophages

The in vitro effect of superoxide anion and lysosomal enzyme activity on the killing of Listeria monocytogenes EGD (listeria) by peritoneal macrophages (PM) was investigated. Generation of superoxide anion by PM stimulated with phorbol myristate acetate (PMA) was significantly increased by intraperitoneal injection of Lactobacillus casei YIT 9018 (LC9018) or Corynebacterium parvum (CP), but not by injection of peptone. However, superoxide anion generation by LC9018-elicited PM stimulated with listeria was not increased any more than that by peptone-elicited PM, and generation of superoxide anion by the PM was affected by the difference in stimuli. The killing of listeria by LC9018- or CP-elicited PM in vitro was significantly less than that by peptone-elicited PM or resident PM. Significant correlation was observed between the anti-listerial activity of PM and the intracellular killing of listeria by PM. On the other hand, beta-glucuronidase and beta-N-acetyl-D-glucosaminidase activities of LC9018-elicited, CP-elicited, or resident PM were significantly weaker than those of peptone-elicited PM, and no significant correlation was observed between the increase in beta-glucuronidase and beta-N-acetyl-D-glucosaminidase activities and the increase in anti-listerial activity. These results suggest that increase in the activity of lysosomal enzymes such as beta-glucuronidase and beta-N-acetyl-D-glucosaminidase is not correlated with the anti-listerial activity of PM, and that superoxide anion has very little effect on the anti-listerial activity of PM in vitro.     

大脑皮质星形胶质细胞体外条件化培养体系的建立

目的：建立大脑皮质星形胶质细胞体外条件化培养体系。 方法：用快速灵敏的MTT比色法测定细胞活力,对纯化细胞进行形态学观察、免疫组化鉴定。结果：经分离、纯化,获得了纯的单一细胞群,星形细胞MTT测定的A值为0.255±0.012。 结论：建立大脑皮质星形胶质细胞体外条件培养体系可以快速、高效得到纯度高的贴壁的星形细胞。     

CASE REPORT: Fibrosing cholestatic hepatitis after living related-donor renal transplantation

A 43-year-old man underwent living related-donor renal transplantation because of chronic renal failure in 1991. During the transplant period, both donor and recipient were seronegative for hepatitis B surface antigen (HBsAg). The donor was seropositive for antibody to hepatitis B surface antigen (anti-HBs) due to hepatitis B virus (HBV) vaccination. After transplantation, FK506 and methylprednisolone had been administered to the patient as immunosuppressants. In 1993, HBsAg appeared in his serum. His alanine aminotransferase level elevated gradually during 1995 and then in 1996, general fatigue, ascites and jaundice developed. At this time his serum was positive for hepatitis B e antibody, contained more than 100000 Meq/mL HBV-DNA and 100% precore mutant. Despite subsequent intensive therapy, liver dysfunction progressed and this patient died of hepatic failure 2 months following admission. At autopsy, the liver exhibited cholestasis, fibrosis extending from the portal tracts, mild inflammation and hepatocytes with a ground-glass appearance. In addition, HBsAg and hepatitis B core antigens had accumulated in the hepatocytes. Consequently, the final diagnosis was fibrosing cholestatic hepatitis (FCH) due to precore mutant HBV infection contracted after renal transplantation. It is unclear when and where the recipient liver became HBV infected. Nevertheless, after renal transplantation, while receiving immunosuppressive drugs, HBV appeared to have the potential to cause hepatic failure and FCH may have been a fatal complication for the recipient.