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Biomedical subjects

R Kasukawa

Publications and source records attributed to R Kasukawa.

At least 91 records · Page 5Linked to original sources
PubMed1997

Cancer cell cycle and adoptive immunotherapy using lymphokine-activated killer cells.

The mechanism underlying the lymphokine-activated killer (LAK) cell cytotoxicity to tumor cells in LAK therapy was studied using flow cytometric cell cycle analysis of Daudi cells. It was suggested that the degree of LAK cell cytotoxicity varied depending upon the phase of the cell cycle that the Daudi cells were in: the resting (G0)-phase/preparation for DNA synthesis (G1)-phase, the DNA synthesis (S)-phase or the preparation for mitosis (G2)-phase/mitosis (M)-phase. The levels of expression of intercellular adhesion molecule-1 (ICAM-1), lymphocyte function-associated antigen-3 (LFA-3) and fibronectin (FN), surface antigens playing important roles in LAK cell cytotoxicity, in the different phases of the Daudi cell cycle have been investigated. The level of the expression of ICAM-1 and LFA-3 was significantly higher in S-phase cells and G2/M-phase cells relative to that in G0/G1 phase cells. The level of expression of FN, however, was significantly lower in S-phase cells and G2/M-phase cells in relation to that in G0/G1 phase cells. On the basis of these results, it may be inferred that the degree of LAK cell cytotoxicity is closely related to the cell cycle phase that the Daudi cells are in and the level of expression of surface antigens associated with the cell cycle phase. For Daudi cells treated with cis-diamminedichloroplatinum (II) (CDDP) and mitomycin-c (MMC), the enhancement of the degree of LAK cell cytotoxicity was directly related to the decrease in the proportion of cells in the S-phase. It may be suggested, therefore, that the proportion of cells in the S-phase in a tumor serves as an index of the sensitivity of the tumor cells to LAK cell cytotoxicity, and that designing a therapeutic strategy on the basis of the analysis of tumor cell cycle would help to improve the success of LAK therapy.

Antineoplastic Agents↗
PubMed1997

Enhancement of eosinophil survival by lipopolysaccharide through releasing granulocyte-macrophage colony stimulating factor from mononuclear cells from patients with bronchial asthma.

BACKGROUND: The mechanism of bronchoobstruction in asthmatics following inhalation of lipopolysaccharide (LPS) is unclear. OBJECTIVE: In this study we have investigated the effects of the LPS on eosinophil survival rate in stimulating peripheral blood mononuclear cells (PBMCs) from patients with asthma. METHODS: PBMC supernatants from 10 asthmatic patients and 4 healthy subjects were compared for eosinophil survival rate using the same purified eosinophils. Two x 10(6) cells/ml of PBMCs were cultured for 24 hours in RPMI 1640 containing 10% fetal bovine serum along with 10 micrograms/ml of LPS. 10(6) cells/ml of eosinophils were incubated for 96 hours in the presence of PBMC-derived culture supernatants at 75% in volume. RESULTS: The viability of eosinophils incubated with PBMC supernatants from asthmatic patients stimulated with LPS was higher, 62.5% +/- 10.6% (mean +/- SD), than that without LPS, 26.9% +/- 10.8%, and also higher than that of PBMCs from healthy subjects with LPS, 40.0% +/- 15.4%. This increasing activity of asthmatic PBMC stimulated with LPS was markedly inhibited from 72.2% +/- 9.7% to 38.5% +/- 6.8% by addition of mouse anti-human GM-CSF antibody to the PBMC supernatant but not mouse anti-human IL-5 antibody. CONCLUSION: These results suggest that LPS enhances eosinophil survival in asthmatics by increasing the production of GM-CSF from PBMCs.

Adult↗
PubMed1997

Substances reactive with mannose-binding protein (MBP) in sera of patients with rheumatoid arthritis.

OBJECTIVES: In order to evaluate the role of mannose-binding protein (MBP) in rheumatoid arthritis, we characterized MBP-binding substances in sera of patients with this disease. METHODS: An enzyme linked immunosorbent assay (ELISA) was used to detect MBP-binding substances in sera of patients with RA. We applied the sera of two RA patients to an immobilized MBP column, and by means of both ELISA and molecular sieve chromatography, examined the substances that bound to MBP. MBP-MASP complexes were added to the fractions containing the binding substances, and C4-consuming activity was examined. RESULTS: Sera of patients with RA showed stronger MBP binding on ELISA than did those of normal controls. In the case of RA sera, both IgG was IgM-RF eluted from the MBP column, whereas with normal controls, only IgG was obtained. The results of molecular sieve chromatography showed that the binding substances of RA patients consisted of immune complexes containing IgG and IgM-RF. These substances were specifically bound to MBP, and once bound, the MBP-MASP complexes were then able to consume C4. CONCLUSION: MBP binds to immune complexes consisting of IgG and IgM-RF, and probably recognizes either the mannose moiety of IgM-RF or the N-acetyl-glucosamine of agalactosyl IgG. When this occurs in RA patients, the lectin pathway would then be activated.

Antigen-Antibody Complex↗
PubMed1996

Antibodies to E1 and E2/Protein X components of pyruvate dehydrogenase complex in sera of patients with primary biliary cirrhosis.

AIMS/METHODS: Using purified E1 component of pyruvate dehydrogenase complex (PDC) from bovine heart, we measured the levels of anti-E1 antibodies in PBC sera using ELISA and determined the degree of inhibition that these antibodies exerted on E1 enzyme activity. We also estimated levels of anti-E2/Protein X (Pro-X) antibodies in PBC sera using purified E2 and Pro-X of PDC which were copurified with E1. RESULTS/CONCLUSIONS: Anti-E1 antibodies were detected in 87.5% (35/40) of PBC sera. Some of these sera inhibited E1 enzyme activity but inhibition did not correlate with levels of anti-E1 antibodies. A high positive correlation (r = 0.918) was found between levels of anti-E1 and anti-E2/Pro-X antibodies, suggesting that anti-PDC antibody production was stimulated by PDC itself. Levels of IgG class anti-E2/Pro-X antibodies were significantly higher in sera of symptomatic PBC patients than in those of asymptomatic PBC patients. It was also found that patients who were positive for only IgM class anti-E2/Pro-X antibodies had early-stage PBC.

Animals↗
PubMed1996

Interleukin-6 induces proliferation of rat hepatocytes in vivo.

BACKGROUND/AIMS: The aim of this study was to assess the effect of interleukin-6 (IL-6) on the proliferation of hepatocytes and to study the interaction between IL-6 and hepatocyte growth factor (HGF) in vivo. METHODS: IL-6 was injected at a dose of 200 micrograms/mg subcutaneously into rats every day for 14 days. Liver and blood samples were obtained at 1, 3, 7 and 14 days during IL-6 administration. Hepatocyte proliferative activity of sera was measured using 3H-thymidine incorporation into cultured rat hepatocytes. To evaluate the proliferative activity of the hepatocytes in tissue sections, hepatic DNA content and immunostaining of the liver tissue sections for proliferating cell nuclear antigen (PCNA) were performed. Plasma HGF levels were measured using specific EIA. In addition, total RNA was extracted from the liver and expression of HGF mRNA was detected by RT-PCR. RESULTS: The DNA contents of liver taken from IL-6-treated rats were increased during IL-6 administration compared with untreated rats. Sera taken from IL-6-treated rats at various intervals during administration also significantly increased 3H-thymidine incorporation by cultured rat hepatocytes compared with sera from untreated rats, suppressing 3H-thymidine incorporation at day 1 and 3 by anti-HGF antibody. IL-6 itself did not increase 3H-thymidine incorporation. Increased expression of PCNA in these hepatocytes was noted from 1 day after IL-6 administration, and at 14 days, the number of PCNA-positive cells was sevenfold greater than in the livers of untreated rats. However, plasma HGF levels showed a peak at day 1, decreased gradually from day 3, and became undetectable by day 14. HGF mRNA expression in livers of IL-6-treated rats was suppressed from day 3 to day 14 of IL-6 administration. CONCLUSIONS: These data show that IL-6 induces an early phase of liver cell growth in vivo and suggest that an increase level of HGF mediates this effect.

Animals↗
PubMed1996

Protective effect of beraprost sodium, a stable prostacyclin analogue, in development of monocrotaline-induced pulmonary hypertension.

Experimental pulmonary hypertension (PH) was induced by a single injection of monocrotaline (MCT), a pyrrolizidine alkaloid extracted from Crotalaria spectabilis. The effect of beraprost sodium, a stable prostacyclin analogue, on the development of MCT-induced PH in rats was studied. Chronic administration of beraprost sodium at a dose of 30 micrograms/kg/day initiated on the same day as MCT injection decreased the degree of PH determined by weight ratio of right ventricular free wall to that of left ventricle plus septum depending on the duration of administration. Although the injection of prostaglandin E1 (PGE1) at a dose of 200 micrograms/kg/day initiated 1 week after MCT injection did not decrease the degree of PH significantly, beraprost sodium administration at doses of 30 and 100 micrograms/kg/day decreased the degree of PH significantly. The cytoprotective effect of beraprost sodium against endothelial cell (EC) damage is believed to be involved in inhibiting development of PH in MCT-injected rats. The amounts of cytokines such as interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF) produced by alveolar macrophages decreased in accordance with the inhibiting effect of beraprost sodium on development of PH, indicating that beraprost sodium inhibited the development of PH in MCT-injected rats not only through its effect of vasodilation and anti-platelet aggregation in pulmonary circulation but also through its antiinflammatory effects.

Alprostadil↗
PubMed1996

Enhancement of lymphokine-activated killer cell cytotoxicity implicated in the increased expression of surface adhesion molecules on tumor cells treated with anticancer agents.

We investigated mechanisms underlying lymphokine-activated killer (LAK) cell cytotoxicity in terms of intensity of expression of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte-function-associated antigen-3 (LFA-3) on Daudi and KATO-III cells treated with cis-diamminedichloroplatinum (II) (CDDP) and mitomycin-C (MMC). Enhancement (mean, 49.8%) of ICAM-1 or LFA-3 in mRNA and protein expression on treated tumor cells was found by flow cytometry, slot-blot RNA analysis, and reverse transcription-polymerase chain reaction (RT-PCR). Increases in adhesion and cytotoxicity of LAK cells to treated tumor cells were 10.1% and 17.7%, respectively. Results suggested that increased ICAM-1 or LFA-3 expression by low-dose CDDP or MMC binds LAK cells to tumor cells, helping to kill tumor cells. Thus, LAK cell therapy with anticancer agent pretreatment could be useful for treatment of cancer patients.

Antibodies, Monoclonal↗
PubMed1996

"Heterophile" antigen in sera of human recipients of renal allografts.

It was noted that many sera of patients with renal allograft produce distinct precipitation lines in gel diffusion tests with about 20% of infectious mononucleosis sera. The antibodies in infectious mononucleosis sera were of IgM isotope, but, interestingly, they could be removed by guinea pig kidney homogenate, which indicated that the reactions studied were of the Hanganutziu-Deicher rather than of the Paul-Bunnell type. This contention was strengthened by the fact that positive transplantation sera reacted also with standard serum with Hanganutziu-Deicher antibodies. Thus far, the presence of the antigen in the transplantation sera could not be related to the clinical status of the patients, however, the antigen was noted primarily in those sera that did not contain heterophile transplantation antibodies. It was proposed that the antigen detected in the transplantation sera was an altered tissue antigen released from the grafted organ. Besides, interactions between two serum samples from the same patient were noted in immunodiffusion tests. These reactions occurred very seldom and were unrelated to heterophile transplantation antigens or antibodies.

Animals↗
PubMed1996

[Use of the the ED046 kit to analyze serum KL-6 in patients with pneumonitis].

KL-6 is a mucinous glycoprotein expressed on Type 2 pneumonocytes, and serum levels of KL-6 are reported to be abnormally high in patients with interstitial pneumonia. A new assay kit for serum KL-6 (ED046) was used in the evaluation of patients with pneumonitis. To clarify whether KL-6 is a useful marker of pneumonitis activity, 649 subjects were studied, including 185 healthy controls, 187 patients with 3 types of interstitial lung diseases, and 277 patients with 4 types of non-interstitial lung diseases. The serum KL-6 level was significantly higher in the patients with pneumonitis (1285 +/- 1196 U/ml) than in the patients without pneumonitis (307 +/- 232 U/ml). The KL-6 level was also significantly higher in patients with clinically active pneumonitis (1708 +/- 1338 U/ml) than in those with inactive pneumonitis (820 +/- 796 U/ml) (p < 0.0001). Serum KL-6 levels correlated significantly with serum c-reactive protein, lactic dehydrogenase, and PaO2 values. These results suggest that the ED046 assay is useful for measuring KL-6 as a marker of pneumonitis activity.

Adult↗
PubMed1996

Clinical features of patients with juvenile onset mixed connective tissue disease: analysis of data collected in a nationwide collaborative study in Japan.

OBJECTIVE: To analyze the clinical features and outcome of juvenile-onset mixed connective tissue disease (MCTD). METHODS: Clinical and laboratory findings were compared in 2 groups of MCTD patients divided according to age at onset: juvenile onset: under 16 yrs: adult onset: 16 yrs or older). RESULTS: Systemic lupus erythematosus-like symptoms, such as facial erythema, photosensitivity. LE cells, lymphadenopathy, and cellular casts, were more frequent in juvenile onset MCTD than in the adult form of the disease. On the other hand, scleroderma-like symptoms, such as esophageal hypomotility, scleroderma-like lesions evident on skin biopsy, pulmonary involvement, proximal scleroderma, and pitting scars, were less frequent in juvenile onset MCTD than in the adult form. Patients with juvenile onset MCTD more frequently met the classification criteria for systemic lupus erythematosus (SLE) and less frequently met those for progressive systemic sclerosis (SSc), compared to patients with adult onset MCTD. At disease onset, hand edema and stiffness were observed less frequently in juvenile onset MCTD than in the adult form. Furthermore, the mortality rate was lower in the former than in the latter (2.8% vs 8.4%). CONCLUSION: Although previous studies have reported severe symptoms and adverse outcome for juvenile onset MCTD, we conclude from this nationwide study in Japan that patients with juvenile onset MCTD exhibit more SLE-like and fewer SSc-like features and have a relatively favorable outcome.

Adolescent↗
PubMed1996

Thrombin in the synovial fluid of patients with rheumatoid arthritis mediates proliferation of synovial fibroblast-like cells by induction of platelet derived growth factor.

OBJECTIVE: To investigate the clotting and fibrinolytic activities in synovial fluid (SF) from patients with rheumatoid arthritis (RA) and to examine the role of thrombin in synovial hyperplasia. METHODS: We measured the amounts of thrombin-antithrombin-III complex (TAT), antithrombin-III (AT-III), thrombin, plasminogen, alpha 2-plasmin inhibitor (alpha 2-PI), and plasmin-alpha 2-antiplasmin complex (PAP) in SF of 20 patients with RA and 16 patients with osteoarthritis (OA). The proliferative response of synovial fibroblast-like cells to thrombin was measured using [3H] thymidine incorporation. Expression of platelet derived growth factors (PDGF) in conditioned medium was analyzed using a Western blot method, and expression of the mRNA of PDGF and their receptors was analyzed by reverse transcription polymerase chain reaction. RESULTS: The amounts of clotting factors (TAT, AT-III) and fibrinolytic factors (plasminogen, alpha 2-PI, and PAP) were significantly higher in the patients with RA than in patients with OA (p < 0.01). Moreover, SF thrombin concentrations of patients with RA correlated significantly with erythrocyte sedimentation rates (rs = 0.751, p < 0.01) and serum C-reactive protein concentrations (rs = 0.531, p < 0.05). Thrombin exhibits mitogenic activity toward synovial fibroblast-like cells in vitro, and this mitogenic activity is associated with an increase in the expression of mRNA of both PDGF-alpha receptor and PDGF-beta-receptor. CONCLUSION: The high levels of thrombin activity in the SF of patients with RA and strong mitogenic activity of thrombin toward the synovial fibroblast-like cells suggest that thrombin plays an important role in the pathogenesis of RA.

Adult↗
PubMed1996

Expression of heat shock protein on lymphocytes in peripheral blood and synovial fluid from patients with rheumatoid arthritis.

OBJECTIVE: To determine whether heat shock proteins (HSP) are expressed on the surface of peripheral blood (PB) lymphocytes and synovial fluid (SF) lymphocytes from patients with rheumatoid arthritis (RA). METHODS: Immunoprecipitation and flow cytometric analyses using anti-Yersinia enterocolitica HSP60 monoclonal antibodies (Mab) were performed. Phenotypes of surface HSP-positive lymphocytes were analyzed further by 2 color flow cytometry. RESULTS: Anti-Y. enterocolitica HSP60 Mab immunoprecipitated 120, 97, 70, 60, 55, and 50 kDa proteins in extracts of healthy donor PB lymphocytes and of Daudi cells after thermal shock. Among these proteins, the 60 kDa HSP showed the greatest degree of precipitation. Surface HSP-positive lymphocytes were detected in 2 of 16 PB and 1 of 12 SF lymphocyte samples. Longitudinal studies indicated that HSP were expressed on the surface of lymphocytes in the active stage, but not in the improvement stage of the disease. Surface HSP-positive lymphocytes were mainly either [gamma delta +] T cells or [CD19+] B cells. CONCLUSION: HSP were expressed on the surface of PB and SF lymphocytes from patients with RA. Surface HSP-positive lymphocytes might be recognized by lymphocytes since HSP are strongly immunogenic.

Adult↗