The recent problems concerning macrophages (Mφ) and granuloma are summarized as follows.
A Granuloma is a specific form of chronic inflammatory processes, which is defined as an organized collection of Mφ and/or their modified cells. Its basic structures are those of mature granuloma by mature Mφ, epithelioid cell granuloma by epithelioid and Langhans' giant cells and foreign body type granuloma by foreign body giant cells. In the granuloma formation, various cytokines such as MIF, MCF, MFF, IL-1 and TNFa may be necessary for an aggregation of Mφ at the initial stage. Epithelioid cell transformation of Mφ depends on various factors such as cellular immunity, some of immune complexes and MDP as a bacterial cell component. According to epithelioid cell transformation, Mφ may increase the autocrine and mutual adhesive properties instead of phagocytic activity. In some organs such as the liver, a granuloma may consist of heterogeneous Mφ which consist of Kupffer cells, exudative Mφ and monocytes. In addition, the formation of infectious granulomas may be influenced by the functional specificities of each organ.

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Inter-α-trypsin inhibitor is thought to be produced in the liver, and to liberate acid-stable proteinase inhibitor (ASPI) which increases under pathologic conditions and is excretd into the urine as urinary trypsin inhibitor. When immunological investigation were undertaken with specific antibodies against ITI and ASPI, differential expression of each immunoreactivity was observed in the liver and plasma. Intense ASPI immunoreactivity was noted in the Kupffer cells and sparse cells in Glisson's sheath, while weak ITI immunoreactivity was present only in the Kupffer cells.
ELISA studies of liver extracts, in contrast to the plasma, failed to demonstrate any component corresponding to ITI, but did show a component corresponding to ASPI. These results suggest that the components of ITI are produced in Kupffer cells and circulate in a complex form ITI.

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Prostaglandins (PG) are known to induce contraction of ileal and uterine smooth muscles. PG are also involved in platelet aggregation and its inhibition. Our recent analysis of structure-activity relationship of PG revealed common structural features associated with PG actions.
The common structure of PG for induction of ileal contraction was also observed in acetylcholine (Ach), histamine, platelet activating factor (PAF) and leukotriene D₄ (LTD₄) . The common structure of PG for induction of uterine contraction was also found in Ach, PAF and a drenaline. The structure associated with PG's action to inhibit platelet aggregation was also found in adenosine and isoproterenol. The structure for platelet aggregating action of PG was also found PAF, ADP, and adrenaline. All PG examined had both a structure for platelet aggregation and a structure for its inhibition in 3-10 regions of the molecules. The ratio of the number of structures for inhibition of platelet aggregation and that for induction of platelet aggregation was highest for PGI₂ and lowest for thromboxane A₂. Thus, prostaglandin molecules had both structures for inducing and inhibiting platelet aggregation.

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The effects of intraperitoneally administered chitosan and cyclodextrins (CDs) on the generation of superoxide anion (O^-₂) in mouse peritoneal macrophages (MPs) were investigated.
MPs obtained from mice injected chitosan, α-CD or β-CD generated larger amount of O^-₂ during phagocytosis of opsonyzcd zymosan or when stimulated by 12-o-tetradccanoylphorbol-13-acetate than those from mice injected proteose peptone, a widely used elicitor for MPs. Especially, gnerated O^-₂ from β-CDelicited MPs was as strikingly large as that from MPs elicited by lentinan (LN), a strong antitumor polysaccharide. These results suggest the possibility that β-CD may act as a host defense stimulator.
In LN-elicited MPs, striking increase of O^-₂ generation was not inhibited by the protein kinase C (PKC) inhibitor 1- (5-isoquinolinylsulfonyl) -2-methylpipera-zine (H-7), but completely disappeared when the calmodulin (CaM) antagonist N- (6-aminohexyl) -5-chloro-1-naphthalenesulfonamide (W-7) was added In contrast, both H-7 and W-7 reduced the aug_mentation of O^-₂ generation by β-CD, although neither H-7 nor W-7 completely inhibited this augmentation. These results suggest that the augmentation of O^-₂ generation by LN might be due only to the increased activity of CaM in the MPs, but that the augmentation by β-CD might be due to the increased activity of both PKC and CaM in the MPs.

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Localization and characterization of human platelet-derived adhernce-inhibiting factors (AIFs) were studied. Subcellular fractionation experiments showed that AIFs were present in both cytosol and granule fractions of human platelets. When cytosol and granule fractions of human platelets were applied to a Superose 6 column, the adherence inhibiting activity of the cytosol fraction was eluted at two different positions (2, 600 and 480 kDa), whereas that of the granule fraction was eluted as a single peak (2, 600 kDa) . Thrombinstimulated platelets released granular AIF extracellularly without releasing a cytosolic marker.
When the purified AIF was analyzed on SDSPAGE, the 340 kDa protein band and the other large protein bands were observed. Under reducing conditions, AIF was separated into two components (340 and 190 kDa) . The purified AIF inhibited human neutrophil adherence to glass, plastic and type IV collagen-coated plastic, whereas it did not affect neutrophil adherence to fibronectin- or plasma-coated plastic. AIF inhibited neutrophil spreading on glass. Neither monocyte adherence nor monocyte spreading was affected by AIF.
These results indicate that AIF selectively affects neutrophil adherence and spreading among phagocytic cells.

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The adherence of circulating leukocytes to the vascular endothelium is a critical step of inflammatory infiltration. In order to understand the influence of the nasal secretion on vascular endothelium, adherence of neutrophils to monolayers of endothelial cells was assayed using ⁵¹Cr-labeled leukocytes and human mucosal microvascular endothelial cells derived from inferior turbinate. Quantitative analysis of interleukin-1α, β and tumor necrosis factor-α were made in the retention fluid of the maxillary sinus of chronic sinusitis and the nasal secretion of nasal allergy using ELISA kit. The vascular endothelial cells acquired adhesiveness for neutrophils after preincubation with maxillary sinus retention fluids of chronic sinusitis. However, no adhesive effect was found in the nasal secretion of nasal allergy. Quantitative analysis revealed the presence of interleukin-1 and the absence of tumor necrosis factor-α in the retention fluids of chronic sinusitis. The adherence inducing effect was heatlabile and almost neutralized by anti-IL-1β antibody in dose dependent manner.
These findings suggest that the IL-1 in the para-nasal secretion of chronic sinusitis induce the adherence of neutrophil to vascular endothelium and further promoted leukocytes infiltrates in the paranasal sinuses, playing an important role in persistency of chronic sinusitis.

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In patients with rheumatoid arthritis (RA), complement levels in synovial fluids (SF) and relative amounts of C3b receptors (CR1) on polymorphonucler leukocytes (PMN) in SF were studied. C4 levels in SF of RA patients were lower than those it patients with osteoarthritis (OA), whereas C3 levels were similar to those of OA. This finding indicated an activation of complement through classical pathway in the joint fluids.
The CR 1 on PMN which will increase through activation of complement has been presumed to inhibit a further activaion of complement. CR 1 levels on PMN in RA-SF were divided into three groups : low (less than 10), middle (between 10 and 40) and high (more than 40) . Low CR 1 group showed significantly lower CH₅₀ levels in SF compared with those of other two groups. CR 1 expression on RASFPMN is different from those of normal subjects, in which amounts of CR 1 are augmented through activation of complement.
Lack of appropriate CR 1 augmentation observed on RASF-PMN would accelerate complemet activation in the joints and cause lesion in joints of patients with RA.

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The effect of piroxicam, a relatively new nonsteroidal anti-inflammatory drug on proteoglycan metabolism has been studied in normal rabbit chondrocytes and diseased part of human articular cartilage in culture. Therapeutic plasma levels of piroxicam (1×10^-5M) had no effects on ³⁵SO₄ incorporation into glycosaminoglycan and ³H-thymidine uptake in the rabbit cultured chondrocyte during 24 hours incubation. On the other hand, piroxicam at concentrations in a range of 1×10^-6 M to 1×10^-4 M, produced dose dependent decreases of proteoglycan release and caseinase activity increased by interleukin-1 in the culture medium from both the rabbit articular chondrocytes and human diseased cartilage.
These results suggest that piroxicam of pharmacological dose may have an ability to protect cartilages by inhibiting the enzymatic breakdown of matrix proteoglycan without effects on proteoglycan synthesis or proliferation of chondrocytes.

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We have reported previously that PGE₂ evoked an increase in intracellular calcium level ( [Ca²⁺] _i) in mouse osteoblastic cells (Eicosanoids, 3, 157-160) . Here, we investigated the effects of PGE₁ and PGF_2a on cAMP production and [Ca²⁺] _i in comparison with those of PGE₂. In osteoblastic clone, MC3T3-E1 cells, PGE₁ stimulated cAMP production, but has no effect on [Ca²⁺] _i whereas PGF_2a evoked only [Ca²⁺] _i increase. In contrast, PGE₂ not only stimulated cAMP production, but also increased [Ca²⁺] _i. From the Scatchard plot analysis of PGE₂ it is confirmed that there are two classes of PGE₂ binding sites (Kd value, 9.2 nM; binding site, 29 fmole/mg protein, and Kd value, 134 nM; binding site, 148 fmole/mg protein) .
These data indicate the possibility that the dual action of PGE₂ is mediated by distinct receptor systems. As increase in [Ca²⁺] _i was caused by PGF_2a and PGE₂, but not by PGE₁, we investigated the displacement study of [³H] -PGF_2a binding. The displacement capacity of unlabeled PGE₂ was about 1/10 of that of PGF_2a, while that of PGE₁ was very low even at 500-fold excess.
These data suggest that the dual effect of PGE₂ is mediated by distinct receptor systems.

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Ciclosporin (CYA) is now widely used in the prevention/treatment of graft rejection and in the treatment of some human inflammatory diseases.
In this study the effect of CYA on the release of histamine from human basophils and rat peritoneal mast cells induced by various stimulis in vitro was examined.
CYA inhibited the histamine release induced by anti-IgE, calcium ionophore A 23187 or concanavalin A (Con A), however it has no effect on the histamine release induced by 12-o-tetradecanoyl pholbol-13-acetate (TPA) or compound 48/80. Inhibition of histamine release by CYA was not influenced by washing the cells before stimulation. CYA inhibited calcium influx induced by calcium ionophore A 23187, but did not inhibit calcium movement induced by compound 48/80. The inhibitory effect of CYA on histamine release induced by ionophore A 23187 was not reduced by the addition of calmodulin.
These results suggest that the mode of action of CYA on histamine release is related to the inhibition of calcium influx.

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PGE₂-induced phosphatidylinositol (PI) breakdown was investigated in medulla and cortex from normal and puromycin aminonucleoside (AN) rat kidney at the several stages.
PI breakdown as elicited by PGE₂ dose-dependently. PGE₂-induced PI breakdown was significantly suppressed one week after the AN administration, and recovered to normal level three and five weeks. This suppression of PI breakdown was clearly correlated to degree of urinary protein excretion induced by AN.
These results suggest that PGE₂ activates PI breakdown in the rat kidney, and the signal transduction of PGE₂ receptors may be damaged in the nephrotic states.

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Anti-inflammatory effect of iodine-enriched egg was evaluated against ear inflammation produced in mice by arachidonic acid and the following results were obtained. (1) High concentration iodine-enriched egg (287 ppm iodine) suppressed ear edema caused by arachidonic acid-induced inflammation in mice by oral administration for 10 days prior to chemical injury. (2) In addition, low concentration iodine-enriched egg (70 ppm iodine) was also effective by administration orally for 30 days prior to the injury. (3) Nordihydroguaiaretic acid was the most effective to suppress ear edema by topical application. (4) Oral administration of aspirin and ordinary egg (less than 3.5 ppm iodine) were not effective. (5) An increase was induced in serum corticosterone levels by feeding of iodine-enriched egg for 10 days.
These results suggested that iodine-enriched egg suppress ear edema in mice by possibly influencing on arachidonic acid metabolism and pituitary adrenal system.

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The Kampo drugs, Saireito, Ninjinyoueito and Kamikihito, inhibited significantly the development of hind foot swellir g and the progression of articular bone destruction in adjuvant arthritic rats, ten per each, maintained on the diet containing the respective drugs at 0.25% for 10 weeks.
Kamikihito was the most effective, and the inhibition of AA was fairly comparable to that produced by indomethacin in a daily dose of 0.5 mg/kg for the identical period. The data presented suggest that Kampo drugs may be useful for the prophylaxis and treatment of rheumatoid arthritis.

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We examined the effects of antirheumatic drugs including cyclosporin A (CsA), bucillamine (SA 96) and salazosulfapyridine (SASP), on the production of, and the responsiveness to interleukin 1 (IL-1) and interleukin 6 (IL-6) and compared them to those of dexamethasone (DM) .
Our study demonstrated that IL-1 production from human peripheral blood adherent cells was inhibited by antirheumatic drugs in the order of DM SA 96 CsA. There was no significant difference between normal controls and patients with rheumatoid arthritis with regard to the sensitivity to the drugs in vitro. Inhibitory effect of SASP was only elicited to rheumatoid adherent cells. IL-6 production was relatively resistent to the drugs examined, although significant but mild inhibition was observed. In contrast, the responsiveness to IL-1 and IL-6 was not affected by the drugs, except that DM augmented responsiveness to IL-6.
These data suggest that antirheumatic drugs might exhibit their antirheumatic action through inhibition of cytokine production including IL-1 and IL-6.

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