MRL/Mp-lpr/lpr (MRL/l) mice which are lymphoproliferation (lpr) gene-congenic mice of MRL/Mp-⁺/₊ (MRL/n) spontaneously develop glomerulonephritis, arteritis and arthritis characterized by the accumulation of macrophages. In this study we investigated the role of lpr gene expression on the functional activities of resident peritoneal macrophages of MRL strain of mice.
MRL/l mice showed remarkable increase of lysosomal enzyme activities and superoxide release of macrophages compared with those of MRL/n mice. In another lpr congenic mice C57BL/6-lpr/lpr (B6/1) showed, however, neither lupus lesions nor the activation of macrophages in spite of the remarkable lymphoproliferation. These results may suggest the importance of background genes of MRL/Mp-⁺/₊ strain of mice in addition to lpr gene expression for the activation of macrophages and further the development of murine lupus of MRL/l mice.

抄録全体を表示

We studied assay system of simultaneous extraction of thromboxane B₂ (TXB₂), 6-keto-prostaglandin F_1α (6KF₁) and prostaglandin E₂ (PGE₂) . Accordingly, TXB₂, 6KF₁ and PGE₂ were fractionated by simultaneous extraction system using octadecylsilyl silica (ODS) suspention and silica minicolumn (BOND ELUT Si) . These fractionated substances were measuerd by radioimmunoassay (RIA) .
Concentration obtained by this assay system in plasma sample of normal individuals (male 47, female 60) were TXB₂: 14-50pg/ml, 6KF₁: 12-33pg/ml and PGE₂: 8.4pg/ ml (male), 4.4pg/ml (female) respectively.
Therefore, we concluded that measurements of TXB₂, 6KF₁ and PGE₂ by our assay system were superior in smaller amount of sample simpler procedures resulting in mass specimen handling and easy for clinical application compared to other assay systems.

抄録全体を表示

Exacerbation of some autoimmune diseases is thought to be related to the generation of active oxygens by leukocytes. It has been known that a number of leukocytes appear frequently in demyelinating lesions of guinea pig brain induced by experimental allergic encephalomyelitis. The present studies showed that cerebroside sulfulic ester (CSE, sulfatide), a typical component of myelin membranes, stimulated guinea pig leukocytes to generate O₂^-, H₂O₂, and probably ⋅OH. Luminol-independent light emitted from CSE-stimulated cells was well correlated with generation of those active oxygens. Multilamella liposomes, which were made of CSE and lecithin, induced H₂O₂ generation by leukocytes. Incubation of CSE-stimulated cells with myelin membranes emitted more light than those without myelin membranes, suggesting that active oxygens released from CSE-stimulated cells react with the membranes. Further studies with thiobarbituric acid reaction revealed that CSE-stimulated cells induced lipid peroxidation in myelin membranes. These results suggest that active oxygens produced by CSE-stimulated leukocytes may have a hazardous effect on myelin membranes, leading to further demyelination.

抄録全体を表示

Hypoxic pulmonary vasoconstriction (HPV) is known to be an important physiological response to preserve an appropriate ventilation-perfusion ratio in the lung. The precise mechanism of HPV remains to be controversial, but recently the role of prostaglandins for mediating and/or modulating HPV has been emphasized. From this viewpoint, we studied the effect of PAF on HPV in anesthetized, open chest mongrel dogs. HPV was assessed by the change of total pulmonary resistance (TPR, PPA-PLA/QAO) . After the stable condition was established, 15-min exposure to 10% O2 followed by 15-min exposure to 100% O₂ was repeated 6 times.
In control group (N=5), spontaneous potentiation of HPV was observed during the 1st to the 5th hypoxic exposures, but an attenuation was occurred in the 6th hypoxic exposure.
In PAF treatment group (N=5), the 6th hypoxic exposure was performed during continuous infusion of PAF at the dose of 33 ng/kg/min for 30 min (totally 1μg/kg) . This group showed no attenuation but potentiation of HPV in the 6th hypoxic exposure.
Pretreatment with CV-3988 (10mg/kg), a specific antagonist of PAF, blocked the potentiating effect on HPV induced by PAF but pretreatment with OKY-046 (10mg/kg), a specific thromboxane synthetase inhibitor, did not block the effect of PAF.
It is suggested that PAF, at the dose of 1μg/kg, acts as vasoconstrictor even in the condition of elevated pulmonary vascular tone induced by HPV in mongrel dogs.

抄録全体を表示

A systemic administration of aqueous form of peptidoglycan polysaccharide polymer (PG-APS) derived from S. pyogenes induced acute and chronic form of polyarthritis in all the rat strains tested. SHR developed severe arthritis 24 hours after injection and developed very low titer of IgM and IgG antibody to PG-APS whereas WKY appeared to be resistant to polyarthritis but developed high titer of antibody to PG-APS. Congenitally athymic nude rats developed acute polyarthritis and high titer of antibody but developed chronic form of polyarthritis only after multiple injection of PG-APS whereas euthymic rnu/+ rats easily developed severe acute and chronic polyarthritis and developed relatively lower titer of antibody than rnu/rnu rats. There existed good correlation between antibody formation and acute polyarthritis among rnu/rnu and rnu/+ rats whereas no correlation existed between chronic form of polyarthritis and antibody formation.
Acute exacerbation of polyarthritis occured within 24 hours after booster injection of PG-APS but no secondary antibody response were observed. It was thus considered that the pathogenesis of this polyarthritis may be different from that of adjuvantinduced arthritis where T cell may play an important role and whether or not antibody to PG-APS is associated with the development of polyarthritis.

抄録全体を表示

Characterization of the heterogenous synovial cells is pivotal to understand the pathogenesis of rheumatoid arthritis. Three different types of cloned rheumatoid synovial cells; dendritic, macrophage-like and fibroblast-like cells classified by morphology have been cultured for up to 8 months without any significant morphological changes. All three types of the cells were mononuclear adherent cells, and HLA-DR positive when stimulated with γ-interferon. Macrophage-like cells phagocytosed large numbers of carbon particles and fibroblast-like cells did to lesser degree while dendritic cells did not. Spontaneous production of interleukin 1-like factor by three different types of the cells have been detected even after the long-term culture, and the ability was in the following order: dendritic cell> macrophage-like cell>fibroblast-like cell.
The results may suggest that majority of the synovial cells could produce interleukin 1-like factor and were responsible for the bony destruction in rheumatoid arthritis.

抄録全体を表示

Ferritin contents in peripheral blood or synovial fluid leukocytes from 24 RA patients and 14 healthy donors were measured by 2-sites immunoradiometric assay as supernatants after cells with 5 times repetition of freezing and thawing were centrifuged at 3000 r.p.m. for 20 min. Ferritin contents per a single cell (fg/cell) were calculated from the value of measured supernatants divided by the number of cells. Polymorphonuclear cells (PMN) and mononuclear cells (MNC) were separated from huffy coats or synovial fluid with heparin (10 u/ml) and hyaluronidase (20 u/ml) by Conray-Ficoll gradient sedimentation method. Peripheral blood mononuclear cells (1×10⁶ cells/ml; PBM) were suspended in RPMI 1640 medium containing 10% fetal calf serum and cultured in 5% CO₂ incubator at 37°C for 7 days with or without addition of different concentration of ferric citrate ranged from 0.01 to 1 mM. Ferritin contents in PBM were increased with addition of ferric citrate in dose-dependent manner. Ferritin contents in peripheral blood PMN (mean ± SEM=5.3±2.6 fg/cell, n=16) and MNC (9.3±3.3 fg/cell, n=16) of RA patients were not significantly different from that of healthy controls (PMN: 4.1 ±2.6 fg/cell, n=14 and MNC: 11.1±2.7 fg/cell, n=15) . It was, however, found in both groups that ferritin contents in MNC were higher than that in PMN. Ferritin contents in synovial fluid PMN (29.7±9.5 fg/cell, n=16) and MNC (62.4±7.1 fg/cell, n=16) of RA patients were remarkably higher than that in peripheral blood leukocytes.

抄録全体を表示

The cell surface phenotypes and interleukin-2 related function were studied employing lymphocytes from peripheral blood (PB) and synovial fluid (SF) of patients with rheumatoid arthritis (RA) . The numbers of IL-2R positive and Ia positive lymphocytes from peripheral blood of patients with RA were increased when compared to normal blood. Con A activated lymphocytes both from PB and SF in patients with RA produced decreased IL-2. In contrast, unstimulated PB lymphocytes from RA patients, who had active disease and whose disease duration was less than 3 years, showed increased proliferation response to IL-2 (P<0.01) . RA SF lymphocytes showed increased response to IL-2 than PB lymphocytes with RA (P<0.01) . In synovial fluid, numbers of 2H4⁺CD4 cell were decreased. Those abnormalities in T lymphocytes from RA patients appeared to play an important role in pathogenesis of RA.

抄録全体を表示

We have studied the distribution of LM with an average particle size of 0.2μm, a suspension of which is in wide use for parenteral nutrition. We incorporated several drugs into LM and have studied a targeting therapy with these LM-drugs.
In the present study, we performed RI angiography using ^99mTc labeled LM, and investigated the distribution of LM to the inflamed joints of patients with rheumatoid arthritis and atherosclerotic lesions. We are also studying the feasibility of the application of this imaging technique to various disease. The ^99mTc labeled LM accumulated in the atherosclerotic lesion and inflamed joints of patients with RA to a greater degree than in other area. This result suggests that LM can be useful as a carrier of targeting therapy in several atherosclerotic and inflammatory diseases, and that the examination using.

抄録全体を表示

Thromboxane (TX) B₂ is formed by platelets during the isolation of blood plasma. The determined level of TXB₂ in plasma sample does not reflect the quantitative endogenous production of TXA₂. Because of this complication, it is more suitable to determine the level of 11-dehydro-TXB₂, one of the most prominent metabolites of TXB₂ which is not formed in the blood, for the indication of TXA₂ synthesis. Therefore, we attempted to develop a sensitive and reproducible enzymeimmunoassay for measuring 11-dehydro-TXB₂. TXB₂, as heterologous hapten, was coupled with β-D-galactosidase using γ-aminobutylic acid as a spacer. Using this method, measurement of 11-dehydro-TXB₂ within the range of 25-50000 picograms per tube was possible. Crossreactivities for TXB₂, 2, 3-dinor-TXB₂ were 9.3% and 3.8%, respectively. However, the crossreactivity for other prostanoids tested was 0.3%.

抄録全体を表示

Bromelain-induced edema in rats was studied and compared to carrageenin-induced edema. The edema reached a maximum 30 min after Bromelain was injected subcutaneously into the rat paw, then decreased slowly, and had almost disappeared 24 hr later. The bromelain-induced edema was obviously potentiated by the oral administration of captopril, a kininase II inhibitor, 1 hr before and after the bromelain was injected; the carrageenin-induced edema was moderately but not dose-dependently potentiated. Dexamethasone suppressed the bromelain-induced and carrageenin-induced edema, but onset of the effect was slow. Nonsteroidal acidic anti-inflammatory agents (indomethacin, piroxicam, phenylbutazone, mefenamic acid, timegadine, and benoxaprofen), except aspirin, exerted no effect on the bromelain-induced edema, whereas they significantly reduced the carrageenin-induced edema. Nonsteroidal nonacidic anti-inflammatory agents such as emorfazone, aminopyrine, tiaramide, perisoxal, and paracetamol, significantly inhibited both types of edema. Antihistamine, antiserotonin, and diuretic agents did not affect the bromelain-induced edema, but diuretic agents inhibited the carrageenin-induced edema. These results indicate that the bromelain-induced edema may be mediated mainly by bradykinin, but not by metabolites of the arachidonic acid cascade or not by biogenic amines, and that the carrageenin-induced edema is mediated predominantly by prostaglandins. The bromelain-induced edema appears to be a useful model to evaluate the effects of nonsteroidal nonacidic anti-inflammatory agents.

抄録全体を表示

Pirazolac is known as a compound having both anti-inflammatory and immunomodulating activities. We studied the effects of Pirazolac on anti-SRBC response in BALB/c mice (female, 7w) and in MRL/Mp-lpr/lpr (MRL/1) mice (male, 20w) with spontaneously occurring autoimmune diseases. In BALB/c mice, Pirazolac enhanced spleen hemolytic plaque forming cell (HPFC) and rosette forming cell (RFC) productions at 3 or 30 mg/kg p.o. On the other hand it suppressed spleen HPFC productions in MRL/l mice at 3 or 30 mg/kg p.o. T cell subsets analysis showed that the effect of Pirazolac in MRL/l mice was based on the increase in suppressor/cytotoxic T cells (Lyt-1-2⁺) in the spleen cells. To clarify the mechanisms of immunomodulating activities of Pirazolac, the effect of Pirazolac on phagocytosis of peritoneal exudate cells of BALB/c mice was studied using Candida parapsilosis as a target cell. Pirazolac enhanced candidacidal activities at 10^-7-10^-6M. This result suggests that some part of immunomodulating activities of Pirazolac was due to activation of macrophages.

抄録全体を表示