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Biomedical subjects

N Mukaida

Publications and source records attributed to N Mukaida.

192 records · Page 11Linked to original sources
PubMed1986

Discrepancy between the production of interleukin 1, alpha-interferon and granulocytic colony-stimulating activity of human monocytes stimulated by a partially purified human urinary colony-stimulating factor.

A partially purified human urinary colony-stimulating factor (pp-CSF), with a specific activity of 1.0 X 10(6) U/mg protein, was purified by using a stepwise DEAE-cellulose anion exchange chromatography and a molecular sieve high performance liquid chromatography (TSK gel G3000SW) sequentially. Production of three kinds of monokines by human peripheral blood monocytes was tested in vitro, which contained granulocytic colony-stimulating activity (G-CSA), interleukin 1 (IL-1) and interferon alpha (IFN-alpha). Human peripheral blood mononuclear cells were separated by Ficoll-Hypaque density gradient centrifugation, and monocytes were obtained by adherence to dishes. Although pp-CSF stimulated monocytes to produce G-CSA in vitro, it failed to stimulate the IL-1 or IFN-alpha production. The discrepancy between production or release of these three kinds of monokines by human monocytes stimulated with pp-CSF suggests that pp-CSF preferentially stimulates human monocytes to produce G-CSA. To test the effects of pp-CSF on human monocytes in vivo, we performed i.v. infusions of pp-CSF to four volunteers, and we then took peripheral blood monocytes. Compared to before the infusion of pp-CSF, G-CSA production by monocytes was enhanced, while production of IL-1 and IFN-alpha was not enhanced after the infusion of pp-CSF. These results suggest that pp-CSF stimulates monocytes to produce G-CSA but not to produce IL-1 nor IFN-alpha.

Cells, Cultured↗
PubMed1986

Effects of anti-Tac antibody on the response of large granular lymphocytes to interleukin-2.

Large granular lymphocytes (LGL), which consist of a unique population comprising almost all of the natural killer (NK) cell activity, were separated from peripheral blood mononuclear cells by sequential depletion of monocytes and conventional discontinuous Percoll density gradient sedimentation. The LGL populations thus obtained exhibited significant levels of interferon-gamma (IFN-gamma) production and proliferation, as well as augmentation of NK cell activity in response to interleukin-2 (IL-2). Among these IL-2-driven phenomena, only IFN-gamma production was markedly inhibited by the monoclonal anti-Tac antibody, which presumably recognized the IL-2 receptor, whereas the proliferative response and the augmentation of NK cell activity were only minimally affected by the same antibody, even at the higher concentration. The dissociation of the effects of anti-Tac antibody on these IL-2-driven events gives us some insight on the role of IL-2 receptors involved in these immunologically interesting functions of IL-2.

Antibodies, Monoclonal↗
PubMed1985

Interleukin 1 (IL 1)-dependent lymphokine production by human leukemic T cell line HSB.2 subclones.

Cloning of a human T cell leukemic cell line, HSB.2, was performed by a limiting dilution method to obtain clones with high levels of IL 2 production. None of the subclones that were obtained produced IL 2 constitutively, and only a low level of IL 2 was produced by the stimulation of these subclones with phytohemagglutinin (PHA) alone. High levels of IL 2 production (greater than 300 U/ml) were observed in several clones when stimulated with a cocktail of PHA and IL 1. Among them, HSB.2-A7-D2, A7-D9, or C5-B2 subclones, which were selected after cloning twice, were most effective in IL 1-dependent IL 2 production. HSB.2 subclones exhibited IL 1-dependent production of a variety of lymphokines other than IL 2, e.g., interferon-gamma (IFN-gamma), B cell growth factor (BCGF), and colony-stimulating factor (CSF). We observed that subclones with high IL 2-producing capacity tended to produce high levels of IFN-gamma or BCGF as well, while the capacity of CSF production was not parallel to these properties. Although several subclones were found to produce IFN-gamma and BCGF simultaneously with minimal IL 2 activity, no subclones with an exclusive BCGF production were obtained. Furthermore, when supernatants from the stimulated A7-D9 subclone were applied to an Ultro-gel AcA54 gel chromatography, it was revealed that IL 2 activity (m.w. 17K to 18K) and IFN-gamma (40K to 45K) were clearly separated, whereas two peaks of BCGF activity coincided with each peak of IL 2 and IFN-gamma, respectively. On the other hand, CSF activity was eluted at a different peak (30K to 35K). These data indicate that IL 2, IFN-gamma, and CSF activities are based on distinct molecules, whereas BCGF activities are indistinguishable from IL 2 and IFN-gamma. The HSB.2 subclones thus selected will provide a useful model for delineating the mechanism of IL 1-dependent lymphokine(s) production, and are a promising candidate for better lymphokine(s) producers.

Cell Line↗
PubMed1984

Interleukin 2-dependent T cell line acquires responsiveness to phorbol myristate acetate and lipopolysaccharide in the course of long-term culture.

We have observed that CT6 cell line, a murine interleukin 2 (IL 2)-dependent T cell line, was highly responsive to phorbol myristate acetate (PMA) and to a lesser extent but significantly responsive to lipopolysaccharide (LPS) in the short-term proliferation assay. In contrast, two other murine IL 2-dependent cell lines, CTLL-2 and NK clone 7, were totally unresponsive to these stimulants. Even when CT6 was recloned by a limiting dilution, no unresponsive clone to PMA was obtained, whereas several clones unresponsive to LPS were obtained. PMA, unlike to IL 2, could not support a long-term culture of CT6. There are no differences between CT6 and CTLL-2 except that the former possessed asialo GM1 while the latter lacked it. Though it is unknown whether this difference is involved in the mechanism of the response of CT6 to PMA or LPS, these data give caution to us against the use of CT6 for the determination of IL 2 activity contaminated with PMA and LPS.

Animals↗
PubMedDate not supplied

Distinct expression of CCR1 and CCR5 in glomerular and interstitial lesions of human glomerular diseases.

We investigated the presence of CCR1- and CCR5-positive cells immunohistochemically in the kidneys of 38 patients with several renal diseases, including 13 crescentic glomerulonephritis patients. In addition, we determined cell phenotypes of CCR1- and CCR5-positive cells using a dual immunostaining technique. Urinary levels of their ligands, for CCR1 and CCR5; macrophage inflammatory protein (MIP)-1alpha, MIP-1beta and regulated upon activation in normal T cells expressed and secreted (RANTES) were evaluated by enzyme-linked immunosorbent assay. CCR1- and CCR5-positive cells were detected in both glomeruli and interstitium of the diseased kidneys. Using a dual immunostaining technique, these positive cells were CD68-positive macrophages (MPhi) and CD3-positive T cells. The number of CCR1-positive cells in glomeruli was correlated with urinary levels of MIP-1alpha. The number of CCR1-positive cells in the interstitium was correlated with both urinary MIP-1alpha and RANTES levels. CCR1-positive cells in the interstitium remained after glucocorticoid therapy, most of which were MPhi, and were correlated with the intensity of interstitial fibrosis and tubular atrophy. Glomerular CCR5-positive cells were well correlated with extracapillary lesions and urinary MIP-1alpha levels, while interstitial CCR5-positive cells, mainly CD3-positive T cells, were correlated with interstitial lesions and urinary RANTES levels. Renal CCR5-positive cells were dramatically decreased during convalescence induced by glucocorticoids. These results suggest that chemokine receptor signaling may be pivotal for human renal diseases through the recruitment and activation of MPhi and T cells; CCR5-positive cells may participate in glomerular lesions including extracapillary lesions via MIP-1alpha and in interstitial lesions via RANTES. CCR1 may be involved in interstitial lesions in resolving phase after glucocorticoid therapy.

Adolescent↗
PubMedDate not supplied

Spontaneous production of interleukin-8 by human lung cancer cells and its augmentation by tumor necrosis factor alpha and interleukin-1 at protein and mRNA levels.

A cell-to-cell interaction between tumors and host inflammatory cells is important for the subsequent cancer progression or regression. We examined the expression of interleukin-8 (IL-8) mRNAs by 9 human lung cancer cell lines and the influences of cytokines on IL-8 production and its gene expression. Substantial expressions of IL-8 gene were detected in 3 lung cancer cell lines (RERF-LC-OK, Lu-134-A-H, YO-88 cells). Moreover, 4 lung cancer cell lines (RERF-LC-MS, RERF-LC-OK, A549 and YO-88) were used to examine the effects of exogenous cytokines--interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor--on IL-8 production by the cells at protein and gene levels. TNF-alpha and IL-1 beta significantly augmented the levels of mRNA expression for IL-8 and its production. These observations indicate that tumor-derived IL-8 may be important in recruiting inflammatory neutrophils and promoting interaction between lung cancer and inflammatory cells.

Gene Expression Regulation, Neoplastic↗
PubMedDate not supplied

Induction and regulation of IL-8 and MCAF production in human brain tumor cell lines and brain tumor tissues.

In order to elucidate the role of inflammatory cytokines in the central nervous system, we examined the production of two leukocyte chemoattractants, IL-8 and monocyte chemotactic and activating factor (MCAF) in brain tumor cell lines. The glioma cell lines tested exhibited high levels of IL-8 and MCAF mRNA expression upon stimulation with IL-1 or TNF-alpha, while none of the neuroblastoma cell lines expressed these cytokine mRNA. Both IL-8 and MCAF mRNA expression depended on the dose of IL-1 alpha and TNF-alpha and appeared very rapidly, reaching maximal levels at 3-6 hr, with substantial production of these cytokines in the culture supernatants. When various immunosuppressive drugs were tested, glucocorticoids but not other immunosuppressive drugs markedly inhibited the IL-1 or TNF-alpha-induced IL-8 and MCAF mRNA accumulation, suggesting that glucocorticoid is a potent regulator of these inflammatory cytokine production in the neural tissues. In addition, reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of IL-8 and MCAF mRNA expression in resected brain tumor tissues including glioblastoma, astrocytoma grade 2, ependymoma and medulloblastoma, indicating that these inflammatory cytokines are expressed in vivo.

Astrocytoma↗