Inflammation and immune response are two major host defence mechanisms. It seems plausible that preceded inflammatory response may regulate the following immune response. We have studied this possibility using the casein-induced peritoneal inflammation. When antigen (sheep red blood cells, SRBC) were introduced into the peritoneal cavity of mice 3-9 hr after intraperitoneal injection of casein, enhancement of IgM responce in spleen was observed 4 days later. This enhancement was also observed in vitro primary antibody responce using spleen cells and SRBC when early inflammatory peritoneal exuded cells (PEC) were added into the culture but not observed by adding the later PEG. These early PEG were composed mostly of polymorphonaclear leukocytes (PMN), and the purified PMN in early PEC were found to contain an immune enhancing factor. The blood PMN which are origin of exuded PMN, could be activated to synthesize the factor (PMN factor) . This factor acts mainly on T cells and makes them produce lymphokines such as interleukin-2 and also has a pyrogenie activity. Thus this factor resembles interleukin-1 (IL-1) in terms of its biologic activity. Then, the possibility wheter the factor also resembles IL-1 in molecular structure was studied using purified PMN factor from rabbit. The factor had mw. 18500 and pl 7.2, and N terminal amino acid sequence was determined and highly homology with human IL-1 β was observed. Then PMN factor is considered to be a member of IL-1 family. Structure and biologic fenction of IL-1 was also reviewed.

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We have previously reported that an increase of the opioid petide content in the rat incisor pulp induced by cavity formation was markedly inhibited following infusion of Des-Arg⁹- [Leu⁸] -bradykinin, a potent bradykinin (BK) -antagonist, into the pulp cavity and the inhibitory effect of BK-antagonist was reversed by combination with BK. These findings suggested that BK might be a triggerr in the increase of the pulpal opioid peptide content. In the present study, we examined the BK-effect on the opioid peptide content in in vitro experimental system newly designed by us, using whole incisor pulp of the rat, in order to certify this suggestion. The met-enkephalin (ME) -like peptide content was remarkably increased by BK at a concentration of 1μM, but not in higher concentrations. The effect of BK (1μM) was antagonized with Des-Arg⁹- [Leu⁸] -BK (1μM) and was not inhibited by FOY-305 (0.001μM), a trypsin inhibitor. From these results, it was demonstrated that the increase of ME-like peptide content induced by BK was due to activation of an enzyme different from trypsin-like enzyme which was mediated through BK-receptor in the pulp.

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Polymyalgia rheumatica (reported by Barber in 1957) is one of the well established rheumatic diseases with characteristic stiffness and pain bilaterally in the neck, shoulder and the pelvic girdle. It is a fever of unknown origin, refractory to antibiotics.
Patients show no abnormalities on physical and roentgenological examination, muscle derived enzymes on biochemistry, EMG or on pathohistology of muscle-biopsy.
From studies up to the present, Bradykinin, prostaglandin, acetylcholine, catecholamine, potassium, ischemia, compression, changes of pH and osmolarity are known to produce pain.
Fifteen cases have been seen in our clinic over the past 5 years and bradykinin was measured in 12 patients with polymyalgia rheumatica.
The serum level of bradykinin was obtained; 9.6 to 21.0 in normals and 17 to 195pg/ml in the patient-group. The mean was 59.4pg/ml.
This higher level of bradykinin in patients suggests some relation in the mechanism of pain in this disease.

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Pleurisy was induced by intrapleural injection of compound 48/80 (5μg/0.5ml) to Sprague-Dawley rats (male 8-9 weeks old) . Plasma exudation was observed only in the first 20 min period. The residual levels of plasma prekallikrein (pre-K) and high molecular weight (HMW) kininogen (KGN) in the exudate of this period were found to be markedly reduced and that of low molecular weight (LMW) -KGN was reduced by 58%. The sinmilar results were obtained after in vitro incubation of plasma with pleural and peritoneal mast cells stimulated by compound 48/80 or purified mast cells degranulated by this histamine releaser. Amidase activity, cleaving Pro-Phe-Arg-MCA, appeared in peritoneal and pleural mast cells one minute after degranulation by compound 48/80. The activity was not attributable to plasma- or tissue- kallikrein, but to chymotrypsin-like and trypsin-like proteases, since the activity was not completely suppressed by soy bean trypsin inhibitor or aprotinin, but was inhibited to 6% by combination of chymostatin and leupeptin. As the residual levels of pre-K and HMW-KGN were reduced in the exudate, even if measured in the presence of chymostatin and leupeptin, it is supposed that these proteases may have inactivated not only factor XII but also pre-K, HMW-KGN and LMW-KGN.

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The cutaneous concentration of histamine and the specific activities of its degrading enzymes, histamine-N-methyltransferase (HMT) and diamine oxidase (DAO), were examined in the Arthus reaction sites induced in guinea pig skin, and kind of cells infiltrated in the lesional sites was also investigated. Histamine concentration revealed a prominent linear decrease at the early stage to about 15% of the control level, and recovered linearly to 85% 6 hours after the initiation without significant infiltration of mast cells and basophils, and no remarkable change was observed thereafter up to 48 hours. The activity of HMT, a major enzyme in the degradation of histamine having almost 15 times greater activity than that of DAO, sustained the control level till 1 hour, followed by a prominent linear decrease to 35% at 6 hours and to 10% at 48 hours. Another histamine degrading enzyme (DAO) activity increased to about 150% till 1 hour, and decreased linearily like the time course of HMT activity. These changes in enzyme activities seemed to well explain the quantitative change in histamine concentration in the reaction sites. The effect of released histamine in the lesional sites was also discussed.

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Morphology and lysosomal enzyme activity were tested in peritoneal macrophage cultures after stimulation in vivo with anti-inflammatory substance, Aloctin A (Alo A), and compared to the same parameters in normal peritoneal macrophages. Alo A-stimulated macrophages were more adhesive and more spread out than non-treated macrophages. The intracellular level of β-glucuronidase activity of macrophages was markedly increased after stimulation with Alo A, while the extracellular level of β-glucuronidase activity was not significantly different from normal macrophages. We also studies the effect of Alo A on heat-induced hemolysis. Alo A markedly inhibited it. The mechanism of the anti-inflammatory properties of Alo A was discussed.

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We assessed the activity of rnethyltransferase and phospholipase A₂ which often induce cell activation, ethanolaminephosphotransferase (EPT) and cholinephosphotransferase (CPT) which mediate the constituent of structural phospholipids of phosphatidylethanolamine (PE) and phosphatidylcholinc (PC), and base-exchange reactions whose biological and physiological significance has not been clarified, in 36 patients with various inflammatory disorders. In the membranes of both neutrophils and lymphocytes from the patients with Behçet's disease, SLE, RA (in active stages) and severe bacterial infections, methyltransferase and phospholipase A₂ activities and ethanolaminc-base exchange activity were markedly or significantly enhanced. In the patients with severe viral infections, only an increase in phospholipase A₂ activity of lymphocyte membrane was observed. On the other hand, EPT and CPT activities, and serine and cholinebase exchange activities were not significantly changed although choline-exchange activity showed trends to decrease in the leukocyte membranes from the patients who showed the enhancement of ethanolamine-exchange. This study postulates that cthanolamme-exchange activity whose mechanism has not been known may be a precursor for the formation of PE which leads to the cell activation by the production of arachidonic acid (AA cascade) induced by mediation of transmethylation and phospholipase A₂ activity.

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Products derived from arachidonic acid (AA) via both the cyclooxygenase (particularly PGE₂) and lipoxygenase (particularly LTB₄) play a role in inflammation. To clarify the effect of eicosapentaenoic acid (EPA) on inflammation, EPA ethyl ester (80% pure, 240 mg/kg/day) was administered for 4 weeks to the rats. The supplementation of a standard rat diet with EPA caused a significant increase in the formation of LTB₅, which is biologically for less active (at least 30 times) than LTB₄, and a decrease in the synthesis of LTB₄ by A 23187 stimulated leukocytes. The EPA rich diet significantly increase the EPA content of leukocytes phospholipids without altering the content of AA, DHA and linoleic acid.
Then, antiinflammatory effect of EPA was assessed using two models of acute inflammation. In the first model EPA fed rat (240 mg/kg/day for 4 weeks) produced significantly lower concentration of PGE₂ and TXB₂ in inflammatory exudate derived from implantation of carrageenin impregnated sponges. Triene prostaglandin were not detected in the exudate however certain level of LTB₅ were present. In the second model, edema induced by injection of carrageenin into rat paws were significantly reduced in animals fed an EPA rich diet. Supplementation of the diet with EPA could, by reducing the synthesis of PGE₂ and LTB₄ and increasing LTB₅ (less active biologically), offer a novel and nontoxic approach to the modulation of an inflammatory response.
Further in vitro experiment disclosed that EPA was better substrate for 5-lipoxygenase than AA but was less favoured substrate for LTA hydrolase. EPA or its oxygenated metabolites, such as LTA₅ or LTB₅ inhibits LTB₄ formation particularly at the level of LTA hydrolase.

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The purpose of this study was to determine the effect of plasma fibronectin (P-FN) on neutrophil function. For this purpose, the interaction of P-FN with neutrophil was examined in two ways. Neutrophils were obtained from human peripheral blood, and particle binding and ingestion were estimated in reaction mixtures with or without P-FN. (a) Particle binding was ascertained by the rosette-forming method using EA or EAC as indicator cells. (b) Percent ingestion of fluorescent microspheres was assayed by flow cytometry.
The results showed that neither particle binding nor ingestion was promoted by the presence of P-FN in reaction mixtures with test particles. From these results, it is suggested that P-FN may not serve as an opsonin to neutrophils.

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A single or multiple systemic administrations of aqueous forms instead of water-in-oil emulsion of biological active acyl muramyl dipeptides (acyl MDPs) induced acute polyarthritis in euthymic rats and athymic nude rats except for WKY rats.
Histological studies revealed acute exudative inflammatory reactions around joints which consisted of hypertrophy of both synovial villi and tendon sheath, infiltration of polymorphonuclear leukocytes and fibrin deposition of the affected joint spaces. During multiple systemic injections of acyl MDPs, this acute arthritis became chronic one in euthymic rats but not in athymic nude rats. It remains uncertain whether chronic polyarthritis results from repeating flare of acute inflammatory reactions by continuing stimulation of adjuvants or whether the chronicity of the disease may require some immunologic process to exogenous substances such as bacterial fragments or autoantigens modified by exogenous substances, such as type II collagen.

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In order to prove an involvement of arachidonic acid metabolites one must accurately estimate these values in biological samples. Although radioimmunoassay is widely used for this purpose, it is limited to certain metabolites. Therefore we devised a simple HPLC method for arachidonic acid metabolites using a fluorescent derivatizing agent, 9-anthryluiazomethane (ADAM) . Reaction of prostaglandin-metabolites with ADAM was performed at room temperature, the mixture was cleaned up with Sep-PAK Silica and then loaded on HPLC column (ODS) . The method was applied to the measurement of metabolites of prostaglandins in rat stomach homogenate.

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The profile of arachidonic acid metabolism in rat dental pulp was studied with a cell-free system. Conversion of exogenously added radioactive arachidonic acid was measured by using thin-layer chromatography. Normal dental pulp could mainly produce 6-keto-PGF_1α and 12- or 15-HETE like lipoxygenase product. When rat dental pulp was experimentally inflamed by applying bacterial lipopolysaccharide, increase of arachidonic acid metabolizing activity was observed. Especially PGE₂ producing activity was elevated 9.3-fold compared with that of the normal. It was suggested that arachidonic acid metabolites were involved in the development of pulpal inflammation.

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In order to clarify the role of entothelial cell in systemic vascular diseases such as arteriosclerosis, diabetic microangiopathy and vasculitis of collagen diseases, the ability of human umbilical endothelial cell cultured for overnight in serum free media RITC 80-7 in MEM to generate superoxide was investigated by using cytochrome C reduction method. It was found that human endothelial cell cultured for overnight was producing spontaneously half amount of superoxide of human monocyte derived macrophage and was producing with PMA one twelfth amount of superoxide of human monocyte derived macrophage. The mechanism of endothelial cell to generate superoxide are remained unsolved.

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Bredinin (BRD) and Cyclosporin A (CyA) as immunosuppressants, and Picibanil (OK432) as immunostimulant, had different effects on T, B cells, and macrophages respectively. The drugs had different influence on the spleen cell count, when single cell suspension was made. FACSTAR analysis requires hemolysis procedure prior to run the equipment. Cell count of the single cell suspension had remarkable decrease after hemolysis procedure. The drugs decreased cell count in different degree respectively. BRD decreased the number of pan B cells, CyA decreased that of L3T4 (helper T), and OK432 increased that of Lyt 2 (suppressor/killer T) . It was difficult to evaluate effects of the drugs in the whole body level. Since less than half amount of cells was able to be analyzed by FACSTAR. It remains to be disclosed how the FACSTAR data reflects changes of immune responsive cell number in the whole body level.

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An autopsy case of a 44-year-old woman with mixed connective tissue disease (MCTD) is presented. Clinically, PSS-like manifestations were outstanding and the entire course of illness was 14 years. The patient died of cerebral hemorrhage. Autopsy disclosed widespread scleroderma, hypertensive pulmonary vascular disease, and nodular regenerative hyperplasia (NRH) of the liver. Relationships between NRH and collagen diseases including MCTD, drugs administered, portal and pulmonary hypertension are discussed.

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