Rat Peritoneal Exudate Cells Research Articles

Effect of suplatast tosilate (IPD-1151T) on types I-IV allergic reactions

We examined the effect of suplatast tosilate (IPD-1151T), which exhibits a class-specific suppression of IgE antibody production, on types I-IV allergic reactions. 1) Type I reaction: IPD-1151T dose-dependently inhibited 48-hr homologous passive cutaneous anaphylaxis (PCA) when the agent was given p.o. 30 min prior to the antigen challenge. To observe the time course of the inhibitory activity, IPD-1151T in a dose of 50 mg/kg was given orally at various times prior to the antigen challenge. IPD-1151T showed inhibitory activity when it was given at 0.5 to 2 hr prior to the antigen challenge, and the maximum inhibition was found when the IPD-1151T pretreatment was 2-hr before the challenge. IPD-1151T also suppressed the antigen-induced degranulation of mesenteric mast cells and histamine release from peritoneal exudate cells of rats. 2) Type II reaction: Only a high dose of IPD-1151T given orally inhibited reversed cutaneous anaphylaxis in rats, whereas the agent did not affect Forssman shock in guinea pigs. 3) Types III and IV reactions: IPD-1151T neither affected the Arthus reaction in rabbits nor the picryl chloride-induced contact dermatitis and sheep erythrocytes-induced footpad reaction in mice. The results obtained here indicate that IPD-1151T shows a relatively specific suppression of the type I allergic reaction with the inhibition of degranulation and histamine release from mast cells.

O-023 - Influence of histamine receptor agonists and antagonists on anaphylactic histamine release from rat peritoneal exudate cells and dispersed human lung mast cells

O-023 - Influence of histamine receptor agonists and antagonists on anaphylactic histamine release from rat peritoneal exudate cells and dispersed human lung mast cells

Inhibitory effects of some natural products on the activation of hyaluronidase and their anti-allergic actions.

The anti-allergic drugs disodium cromoglycate (DSCG) and tranilast are strong inhibitors of hyaluronidase. In the development of new anti-allergic drugs, we studied the inhibitory effects of some natural products on the activation of hyaluronidase. Among the compounds tested, liquiritigenin, isoliquiritigenin and baicalein showed dose-related inhibitory effects. Anti-allergic activities of these compounds were evident from the facts that they inhibited the histamine release from rat peritoneal exudate cells induced by antigen, compound 48/80 and calcium ionophore A-23187, and from their inhibitory effect on Shultz-Dale reaction using sensitized guinea pig ileum.

PSORALEN‐PHOTOSENSITIZED DAMAGE OF RAT PERITONEAL EXUDATE CELLS

Psoralen-sensitized photodamage (PUVA) of rat peritoneal exudate cells was investigated. Quartz-activated luminol-dependent chemiluminescence (ChL) was registered and the amount of trypan-positive cells was determined. Irradiation of peritoneal exudate cells in the presence of psoralen resulted in a dose-dependent monotonous inhibition of ChL. The reciprocity law of irradiation intensity and duration of irradiation was not valid for the observed inhibition of ChL: the inhibition increased with higher intensity. When psoralen previously photooxidized in ethanol (POP) was added to peritoneal exudate cell suspension, a double-phase response depending on psoralen irradiation dose was obtained: ChL activation was observed at low doses of UVA, ChL inhibition at high doses. Chemiluminescence inhibition correlated well with the increase in the number of trypan-positive cells. It may be supposed that the observed effects of PUVA or POP treatment are caused by cell cytoplasmic membrane damage.

PSORALEN-PHOTOSENSITIZED DAMAGE OF RAT PERITONEAL EXUDATE CELLS

Abstract— Psoralen-sensitized photodamage (PUVA) of rat peritoneal exudate cells was investigated. Quartz-activated luminol-dependent chemiluminescence (ChL) was registered and the amount of trypan-positive cells was determined. Irradiation of peritoneal exudate cells in the presence of psoralen resulted in a dose-dependent monotonous inhibition of ChL. The reciprocity law of irradiation intensity and duration of irradiation was not valid for the observed inhibition of ChL; the inhibition increased with higher intensity. When psoralen previously photooxidized in ethanol (POP) was added to peritoneal exudate cell suspension, a double-phase response depending on psoralen irradiation dose was obtained: ChL activation was observed at low doses of UVA, ChL inhibition at high doses. Chemiluminescence inhibition correlated well with the increase in the number of trypan-positive cells. It may be supposed that the observed effects of PUVA or POP treatment are caused by cell cytoplasmic membrane damage.

Effect of anticancer drugs on the release of tumour necrosis factor in vitro.

The anticancer drugs Adriamycin, bleomycin, vincristine, 4-hydroperoxycyclophosphamide, FK156 and FK565 and the immunosuppressive drug cyclosporin-A were tested for their effect on lipopolysaccharide-induced tumour necrosis factor-alpha (TNF) release from rat peritoneal exudate cells in vitro. FK565 resulted in a marked enhancement of TNF release when added to cultures at doses of 1 microgram/ml and 10 micrograms/ml. However, FK156, Adriamycin, bleomycin, 4-hydroperoxycyclophosphamide, vincristine and cyclosporin A did not affect the release of TNF under the same conditions. Puromycin inhibited the release of TNF in the same system at non-cytotoxic doses of 1 microgram/ml and 10 micrograms/ml.

Cellular immune responses of the rat to experimental infection with Dermatophilus congolensis

The host cell-mediated immune response was examined following experimentally-induced infection of rats with Dermatophilus congolensis, the causal agent of the skin disease dermatophilosis. Mononuclear cells (MC) isolated from Wistar rats 10 days following the induction of a third infection underwent a strong and specific proliferative response, as assessed by a [ 3H]thymidine incorporation assay, when cultured with various concentrations of inactivated D. congolensis cocci. Using specific monoclonal antibodies in an indirect fluorescent antibody test, this in vitro response was found to be characterised by a large expansion of the W3/25 (T-helper phenotype) population to form 56% of the total. Finally, the primed and stimulated MC were assessed for their ability to produce factors capable of inhibiting macrophage migration. The culture supernatants of D. congolensis-stimulated MC from infected rats caused significant migration inhibition of normal rat peritoneal exudate cells, whilst the supernatants of similarly-stimulated MC from naive rats failed to cause significant inhibition. The results show that a MC subpopulation becomes primed following experimentally-induced infection with D. congolensis and becomes activated after subsequent, in vitro, exposure.

Effect of dietary oils on the production of n−3 and n−6 metabolites of leukocyte 5-lipoxygenase in five rat strains

We examined the influence of various dietary oils, including linseed and fish oil on the relative rates of leukotriene B 4 (LTB 4) and LTB 5 production by rat peritoneal exudate cells in five rat strains. While there was an association between the membrane phospholipid levels of the fatty acid precursors (arachidonic acid (AA) and eicosapentaenoic acid (EPA)) and the rate of synthesis of their respective 5-lipoxygenase products (LTB 4 and LTB 5), the rate of LTB 4 synthesis was a combined function of both AA and EPA levels. We observed a strong linear relationship (correlation coefficient = 0.99) between the ratio of EPA/AA in the cell membrane phospholipids and the ratio of LTB 5/LTB 4 produced by these cells in vitro; this association was independent of genetic (strain) variability and was independent of the source of EPA (dietary EPA or EPA endogenously synthesized from dietary α-linolenic acid).

Potent Inhibitory Activity of HSR-6071, a New Antiallergic Agent, on Passive Cutaneous Anaphylaxis (PCA)

Antiallergic effects of 6-(1 -pyrrolidinyl)-N-(1H-tetrazol-5-yl)-2-pyraz-inecarboxamide (HSR-6071), a newly synthesized agent, were investigated. The 48-hr homologous passive cutaneous anaphylaxis (PCA) in rats was inhibited in a dose-dependent manner by i.v. and p.o. administration of the agent (ED50=0.0096 mg/kg and 0.18 mg/kg, respectively). The IgE-mediated histamine release from rat peritoneal exudate cells was inhibited by HSR-6071, with an IC50 of 4.6×10−10 M. Regarding the non-immunological histamine release, HSR-6071 inhibited compound 48/80-induced, but not A23187-induced and spontaneous histamine release. On the other hand, an increase in vascular permeability induced by histamine, serotonin and bradykinin was unaffected by HSR-6071 in doses sufficient to inhibit PCA. In addition, the contractile responses of isolated guinea pig ileum to histamine, acetylcholine and serotonin were also unaffected by the agent even in a high concentration of 10−4 M. These results indicate that HSR-6071 possesses a potent antiallergic activity and that the inhibition of PCA by HSR-6071 may be due to the suppression of chemical mediators release from mast cells.

Potent inhibitory activity of HSR-6071, a new antiallergic agent, on Passive Cutaneous Anaphylaxis(PCA).

Antiallergic effects of 6-(1-pyrrolidinyl)-N-(1H-tetrazol-5-yl)-2-pyrazinecarboxamide (HSR-6071), a newly synthesized agent, were investigated. The 48-hr homologous passive cutaneous anaphylaxis (PCA) in rats was inhibited in a dose-dependent manner by i.v. and p.o. administration of the agent (ED50 = 0.0096 mg/kg and 0.18 mg/kg, respectively). The IgE-mediated histamine release from rat peritoneal exudate cells was inhibited by HSR-6071, with an IC50 of 4.6 x 10(-10) M. Regarding the non-immunological histamine release, HSR-6071 inhibited compound 48/80-induced, but not A23187-induced and spontaneous histamine release. On the other hand, an increase in vascular permeability induced by histamine, serotonin and bradykinin was unaffected by HSR-6071 in doses sufficient to inhibit PCA. In addition, the contractile responses of isolated guinea pig ileum to histamine, acetylcholine and serotonin were also unaffected by the agent even in a high concentration of 10(-4) M. These results indicate that HSR-6071 possesses a potent antiallergic activity and that the inhibition of PCA by HSR-6071 may be due to the suppression of chemical mediators release from mast cells.

特異的Ｔｈｒｏｍｂｏｘａｎｅ（ＴＸ）Ａ２合成酵素阻害剤（Ｅ）‐３‐〔ｐ‐（１Ｈ‐ｉｍｉｄａｚｏｌ‐１‐ｙｌｍｅｔｈｙｌ）ｐｈｅｎｙｌ〕‐２‐ｐｒｏｐｅｎｏｉｃ ａｃｉｄ（ＯＫＹ‐０４６）の抗アルルギー作用 （第１報） Ｉ型アレルギー反応に対する作用

We studied the effects of OKY-046 on type I allergic reactions. OKY-046 (100 mg/kg) given orally suppressed antigen-induced bronchoconstriction and TXB2 generation in broncho alveolar lavage fluid in rats passively sensitized with anti-DNP-As monoclonal IgE. At the dose of 30 mg/kg given intraduodenally, it also inhibited antigen-induced bronchoconstriction in guinea pigs passively sensitized with anti DNP-As serum and actively sensitized with ovalbumin. However, aspirin (30 mg/kg) didn't suppressed them significantly. Azelastine (10 mg/kg) inhibited bronchoconstriction in passively sensitized rats and actively sensitized guinea pigs. In 48 hour homologous PCA reactions of rats and mice, oral administration of OKY-046 (300 mg/kg) and tranilast (100 mg/kg) suppressed the extravasated dye in the skin. OKY-046 decreased histamine release from passively sensitized rat peritoneal exudate cells. There was no effect of OKY-046 on SRS-A and leukotriene release from actively sensitized guinea pig lungs and passively sensitized rats. In conclusion, we think that OKY-046 should be an useful asthmatic drug or anti-allergic drug by oral administration.

Structure of the Rat Interleukin 6 Gene and Its Expression in Macrophage-derived Cells

Rat peritoneal exudate cells produce two interleukin 6 (IL6) messenger RNA species, a major 1200 nucleotide and a 5-fold less abundant, 2400-nucleotide species. A cDNA clone representing the major species was isolated, and sequenced. The 1055-base pair insert covered the 3'-nontranslated region, the 211 triplet coding region and most of the 5'-nontranslated region. The derived rat IL6 amino acid sequence was 93 and 58% identical, respectively, with mature murine and human IL6. Rat IL6 lacks N-glycosylation sites but contains a fifth cysteinyl residue in addition to the 4 residues shared in conserved positions with murine and human IL6. Stable murine L cell and human HeLa-derived cell lines were established by cotransfection with rat IL6 cDNA and a selectable neomycin resistance marker. These lines secrete 9-fold increased amounts of functional IL6 over their respective parental cells. A stable rat macrophage-derived cell line, RM-SV1, was established by transformation with simian virus 40. IL6 and Il1 mRNA levels are inducible 20- and 3.5-fold, respectively, in this line by treatment with lipopolysaccharide with kinetics characteristic of macrophages. A set of three overlapping genomic DNA clones was isolated and a 10-kilobase DNA segment was sequenced containing the rat IL6 gene plus 2.9 kilobases of 5'-flanking and 1.3 kilobases of 3'-flanking sequences. The two transcription start sites used in RM-SV1 cells were mapped within 5 base pairs of each other. The exon/intron boundaries are conserved with the murine and human IL6 genes. The two IL6 mRNA species are generated by alternative polyadenylation at sites separated by a distance of 1.2 kilobases. The intervening region contains a repetitive element 72-80% identical with the rat and murine consensus L1 family sequences.

Effect of anticancer agents neothramycin, aclacinomycin, FK-565 and FK-156 on the release of interleukin-2 and interleukin-1 in vitro.

Four anticancer agents, neothramycin, aclacinomycin, FK-565 and FK-156, were tested for their effects on concanavalin-A-induced interleukin-2 release from rat splenocytes in vitro. Neothramycin showed an enhancement of the release of interleukin-2, whereas aclacinomycin had no effect. FK-565 and FK-156 were found to inhibit the release of interleukin-2 under similar conditions. The inhibition was much more marked with FK-565. These drugs were also tested for their effects on the release of interleukin-1 from rat peritoneal exudate cells stimulated by lipopolysaccharide in vitro. Neothramycin and aclacinomycin did not affect the release of interleukin-1; however, both FK-565 and FK-156 resulted in its enhanced release under these conditions.

Mechanism f inhibitory action of tranilast on the release of slow reacting substance of anaphylaxis (SRS-A) in vitro: Effect of tranilast on the release of arachidonic acid and its metabolites.

We investigated the mechanism of inhibitory action of tranilast, one of the anti-allergic drugs, on the release of slow reacting substance of anaphylaxis (SRS-A). Ionophore A23187 (0.5 or 0.2 micrograms/ml)-induced SRS-A release from rat peritoneal exudate cells (PEC) or human leucocytes was inhibited by tranilast (10(-5)-10(-3) M). The IC50 (the concentration which gives 50% inhibition) of tranilast on these reactions was approx. 10(-4) M. Prostaglandin (PG)E2 release from sensitized purified rat mast cells (PMC) by a specific antigen (DNP-Ascaris) was markedly suppressed by tranilast (10(-3) M). Similarly, ionophore A23187-induced PGE2 and 6-keto-PGF1 alpha releases from rat PEC were inhibited by tranilast (10(-5)-10(-3) M). DNP-Ascaris antigen-induced 3H-arachidonic acid (AA), 3H-PGE2, 3H-PGF2 alpha and 3H-PGD2 releases from rat PMC were markedly suppressed by tranilast (10(-5)-10(-3) M), DSCG (10(-5)-10(-4) M) and mepacrine (10(-3) M). The activity of AA-converting enzymes such as 5-lipoxygenase, cyclooxygenase, PGI2 synthetase, and glutathione-S-transferase was hardly influenced by tranilast (10(-5)-10(-3) M). From these results, we suggest that the mechanism of the inhibitory action of tranilast on the release of SRS-A is related to the processes prior to dissociation of AA from the membrane phospholipids.

Mechanism of Inhibitory Action of Tranilast on the Release of Slow Reacting Substance of Anaphylaxis (SRS-A) In Vitro: Effect of Tranilast on the Release of Arachidonic Acid and Its Metabolites

We investigated the mechanism of inhibitory action of tranilast, one of the anti-allergic drugs, on the release of slow reacting substance of anaphylaxis (SRS-A). lonophore A23187 (0.5 or 0.2 μg/ml)-induced SRS-A release from rat peritoneal exudate cells (PEC) or human leucocytes was inhibited by tranilast (10-5–10-3 M). The IC50 (the concentration which gives 50% inhibition) of tranilast on these reactions was approx. 10-4 M. Prostaglandin (PG)E2 release from sensitized purified rat mast cells (PMC) by a specific antigen (DNP-Ascaris) was markedly suppressed by tranilast (10-3 M). Similarly, ionophore A231 87-induced PGE2 and 6-keto-PGF1α releases from rat PEC were inhibited by tranilast (10-5–10-3 M). DNP-Ascaris antigen-induced 3H-arachidonic acid (AA), 3H-PGE2, 3H-PGF2α and 3H-PGD2 releases from rat PMC were markedly suppressed by tranilast (10-5–10-3 M), DSCG (10-5–10-4 M) and mepacrine (10-3 M). The activity of AA-converting enzymes such as 5-lipoxygenase, cyclooxygenase, PGI2 synthetase, and glutathione-S-transferase was hardly influenced by tranilast (10-5–10-3 M). From these results, we suggest that the mechanism of the inhibitory action of tranilast on the release of SRS-A is related to the processes prior to dissociation of AA from the membrane phospholipids.

Iron deficiency and interleukin 1 production by rat leukocytes

The production of the immunotransmitter, interleukin 1 (IL-1), by peritoneal exudate cells (PEC) was examined in iron-deficient and control rats. Three groups of weanling male Sprague-Dawley rats were fed a purified diet containing 6, 12, or 35 ppm iron for 6 wk to produce severe iron deficiency, moderate iron deficiency, or adequate iron status. Crude IL-1 samples were prepared from acute PEC and assayed for activity. IL-1 preparations from severely and moderately iron-deficient rats enhanced mouse thymocyte proliferation in vitro less than half as much as IL-1 preparations from control rats. In a rabbit bioassay, injection of IL-1 prepared with PEC from either group of iron-deficient rats had little effect on body temperature or plasma minerals, while IL-1 from iron-adequate source PEC produced a febrile response and markedly lowered plasma iron and zinc in recipient rabbits. Severe or moderate iron deficiency, then, clearly impairs IL-1 production by rat PEC.

Oxatomide inhibits the release of bronchoconstrictor arachidonic acid metabolites (iLTC4 and PGD2) from rat mast cells and guinea-pig lung

The effect of oxatomide, an orally active antiallergic drug, on immunoreactive LTC4 (iLTC4) production has been studied in rat peritoneal exudate cells (PEC) and guinea-pig lung fragments using the calcium ionophore A23187 and specific antigen in vitro. Oxatomide (10(-5) M) inhibited iLTC4 release by 70% with A23187 from rat PEC, and by 48% with antigen from guinea-pig lung. Oxatomide is supposed to affect the biosynthesis pathway of leukotrienes, because oxatomide inhibits 5-lipoxygenase from guinea-pig peritoneal leukocytes with an IC50 17 microM. Oxatomide also depressed the release of PGD2 from rat peritoneal mast cells stimulated by A23187 (IC50 4.2 microM). The effects of oxatomide on iLTC4 and PGD2 release were more potent than other antiallergic drugs (DSCG, ketotifen, tranilast).

受身皮膚アナフィラキシーモデルにおけるＫＰ‐１３６のアレルギー反応抑制作用

The pharmacological properties of KP-136 were studied using cutaneous reactions in rats and guinea pigs. KP-136 remarkably inhibited the passive cutaneous anaphylaxis (PCA) with intravenous and oral dosing. However, the inhibitory effect of KP-136 had an apparent species difference. Thus, KP-136 was more effective on rat PCA than that of guinea pig. In four rat cutaneous reactions produced by three allergic reactions and compound 48/80, KP-136 was remarkably effective on two homologous PCA induced by IgE and IgGa. The intravenous and oral doses for 50% inhibition on the PCA were 0.2 mg/kg to 0.4 mg/kg and 0.5 mg/kg to 0.9 mg/kg, respectively. KP-136 was scarcely effective on cutaneous reactions elicited by intradermal injection of histamine and serotonin which are main chemical mediators in rat homologous PCA. However, KP-136 blocked the degranulation of mast cells and decrease of histamine content in skin elicited by the PCA. In addition, KP-136 showed a potent inhibition on the immunological release of histamine from rat peritoneal exudate cells. The concentration for 50% inhibition on the histamine release was 5 ng/ml. These findings indicate that KP-136 is an oral potent inhibitor on PCA, and it acts by blocking the release of chemical mediator(s) from mast cells.

Enzymatic study to characterize the slow reacting substance of anaphylaxis (SRS-A) and leukotrienes.

Slow reacting substance of anaphylaxis (SRS-A) has been shown to be one of the major mediators in hypersensitive reactions and to be composed of leukotriene (LT) C4, LTD4 and LTE4. In the present study, we examined the properties of SRS-A released from sensitized guinea pig lungs by antigen and SRS released from rat peritoneal exudate cells and from human leucocytes by ionophore A23187 (0.5 and 0.2 microgram/ml, respectively). By the incubation with SRS-A, SRS and LTs with arylsulfatase (type V) in pH 5.7 buffered solution at 37 degrees C for 30 min, SRS-A and LTD4 were greatly inactivated and rat SRS was slightly inactivated, but human SRS and LTC4 were not inactivated at all. The same results were obtained when aminopeptidase was used in place of arylsulfatase. Moreover, when SRS-A, LTC4 and LTD4 were incubated with 0.02 mg/ml of gamma-glutamyltranspeptidase (gamma-GTP) pH 8.0 buffered solution at 37 degrees C for 30 min, the activities of SRS-A and LTD4 were slightly decreased, but those of SRS and LTC4 were obviously potentiated. On the other hand, incubation with a large amount of gamma-GTP (0.2 mg/ml) a dose at which this enzyme preparation showed clear aminopeptidase activity, SRS-A, SRS, LTC4 and LTD4 were obviously inactivated. In addition, we found a peak of LTD4 in guinea pig SRS-A, that of LTC4 in human SRS, and that of LTC4 in rat SRS on high performance liquid chromatograms. From these results, we demonstrated that guinea pig lung SRS-A is mainly composed of LTD4, human leukocyte SRS is mainly LTC4, and rat peritoneal SRS is composed of both LTC4 and LTD4. The inactivation of LTD4 and SRS-A by arylsulfatase may be due to aminopeptidase contamination in the enzyme preparation.

Specific suppression of antigen-antibody reactions by a dialysate from Dermatophagoides farinae.

Mite dialysate prepared from an extract of house dust mite, Dermatophagoides farinae, suppressed 48-h homologous passive cutaneous anaphylaxis (PCA) in rats caused by a non-dialyzable fraction of the mite extract (mite antigen), and the suppression was dose-dependent with a high specificity for the antigen. The mite dialysate itself slightly elicited the PCA. Fraction 3 obtained from the mite dialysate by means of DEAE-Sephadex A-25 column chromatography suppressed PCA in rats and guinea pigs more potently than mite dialysate. Histamine release from sensitized rat peritoneal exudate cells (PEC) induced by the mite antigen was suppressed in a dose-dependent manner when the PEC had been previously incubated with fraction 3. However, incubation of the PEC with fraction 3 caused a significant release of histamine compared to the spontaneous value. Fraction 3 clearly inhibited the passive hemagglutination of the mite antigen-conjugated sheep red blood cells caused by rat hyperimmunized serum. From these results it was considered that certain haptenic substances might be present in the mite dialysate.