Chemotaxis of neutrophils or macrophages is a complex behavioral response to chemoattractant and one of the more important processes by which cells move from the blood vessels to an inflammatory site in the tissues. In this review, I will discuss about recent progresses in the field of research for mechanisms of chemotaxis first. The discovery of formyl peptides as chemoattractant by Dr. E. Schiffmann has given us many new knowledge about the mechanisms of chemotaxis. Highly radioactive peptides were used to demonstrate and study the peptide receptors on the human and the animal neutrophils or macrophages. The receptors on the cell membrane mediated both chemotaxis and release of lysosomal enzymes. Post receptor events which occur subsequent to the interaction of attractant with receptor are not clear yet. But the internalization of receptor-attractant complex into the cell, the involvement of methylation reaction and transglutaminase appear to be important way of regulating chemotaxis. Secondary, agents that are not chemoattractants themselves but that regulate cell movement are termed modulators. I am interested in the phenomenon that tumors show depressed immunity and susceptibility to infection. Many studies indicate that tumors produce substances that depress accumulation of cells to the site of tumors for the protection of themselves. To demonstrate such modulator is also essential for the study of inflammatory process.

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Adjuvant-induced arthritis (AA) can be produced by a single injection of Freund's complete adjuvant (FCA) containing Mycobacterium. Various bacteria, their cell walls and their peptidoglycans can be substituted for Mycobacterium in FCA. Asynthetic adjuvant, N-acetylmuramyl-L-Ala-D-isoGln (MDP) also produced polyarthritis indistinguishable from AA in terms of clinical course and chronic granulomatous lessions around joints. MDP produced very severe arthritis in euthymic rats but not in athymic nude rats. We have not yet succeeded in reconstituting the disease susceptibility of nude rats by thymus cells or lymphoid cells. All of the germfree (GF) F344 rats developed severe AA whereas conventional and specific pathogen free (SPF) rats developed the disease with low incidence and less severity. When GF rats were associated orally with E. coli and 4 weeks later were injected with FCA, E. coli-associated GF rats developed very mild disease comparable to that of SPF rats. Lactobacilli-associated GF rats developed much severe disease than that of GF rats. These findings suggested that (i) thymus plays an important role in pro-moting the development of AA; (ii) bacterial flora must exert some effects on the development of AA.

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We developed an efficient method for production of experimental polyarthritis. Native collagen (type II) from human costal cartilage was emulsified with incomplete Freund's adjuvant (IFA) containing the synthetic adjuvant, N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP), and injected into one hind footpad of the PVG/c rat. This method reproducibly induced severe arthritis with high incidence, whereas the collagen alone in IFA produced mild arthritis with low incidence. MDP alone in IFA was shown to be minimally arthritogenic under identical conditions.
MDP and its adjuvant-active analogs stimulated macrophages to produce a soluble factor (T cell activating factor, TFA) which was shown to be efficient in antigeninduced MIF production of immune lymphocytes as well as mitogen-induced proliferation of T lymphocytes. The in vitro system developed in this study should contribute to the understanding of the target cells for adjuvant activity of MDP. Moreover, the Mφ factor produced by stimulation with MDP may be advantageous for characterization of the factor, since MDP of low molecular weight appeared to be easily removed from the culture sup.

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An apparantly nonimmunogenic synthetic compound N, N-dioctadecyl-N', N'-bis (2-hydro-xyethyl) propanediamine (CP-20961) disolved in mineral oil and made into emulsion (10 mg/0.2 ml/rat) induced polyarthritis by single injection in the footpad of Lewis rats. The polyarthritis was almost indistinguishable morphologically from classic adjuvant arthritis induced by Freund's complete adjuvant, a disease generally thought to be the result of a dayed type hypersensitivity reaction to a constituent (s) of the injected tubercle bacilli. The joints demonstrated intense destruction, profound pannus formation, exuberant periostal new bone formation, and large clusters of neutrophils infiltration. The disease induced by alkyldiamine (CP-20961) and that induced by Freund's complete adjuvant followed the same time course and almost identical pattern of development of clinical and histopathological features.

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The effect of treatment of glomerulonephritis in NZB mice with large doses of corticosteroids were examined.
Twenty-two mice were ranged no less than 10 months of age. From 6 months of age, 6 mice were injected intraperitoneally with 1 mg/kg/day of prednisolone and 5 were with 30 mg/kg/day, twice a week.
In the treated group, there were marked reduction of mesangial or paramesangial deposits and other abnormal findings of glomeruli, compared with the controls. These findings were compatible to that of human lupus nephritis treated with “pulse therapy”.
It was conceivable that both anti-inflammatory and immunosuppressant effects of corticosteroid might have prevented the development of overt glomerulonephritis.
It was concluded that these therapeutic studies in NZB mice were worthy to assess on the efficacy of medicinal therapy in glomerulonephritis and it would be offered a promissing new approach to treat established human glomerulonephritis.

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Development of experimental animal models for human diseases has strongly accelerated the advance of basic and clinical studies. MRL/l mice develop lupus erythematosus lesion, vasculitis, arthritis, and lymphoma spontaneously. MRL/l mice possess serological and immunological abnormalities such as antinuclear antibodies, anti-DNA, anti-Sm, rheumatoid factor, cryoglobulins, immune complex, hypergammaglobulinemia, per se.
As for inflammatory lesions, MRL/l mice had subacute glomerulonephritis in which the mesangial cell proliferation was conspicuous. Electron microscopic study revealed the deposition of electron-dense material on the subendothelial side of the glomerular basement membrane and in the mesangial cells. In the fluorescein study, granular deposition of IgG, IgM, and C 3 was noticed along the basement membrane and in the mesangial cells. Vasculitis was seen in entire body. Histopathologically, it was vasculitis characterized by granulation and round cell infiltration. Panarteritis and fibrinoid necrosis were also identified in some sections, indicating that the vasculitis resembled human periarteritis nodosa. The peripheral joints were principal sites of the arthritis seen in MRL/l mice. The histopathological changes of the affected joints resembled one of the histopathological types of human rheumatoid arthritis, i.e. “lympho-plasmocytes reaction accompanied with connective tissue proliferation”. It was presumed the this arthritis might be located between human rheumatoid arthritis and rat adjuaant arthritis.
MRL/l mice will serve as useful and important experimantal animal model to elucidate pathogenesis and establish treatments for inflammation and immunological disorders.

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The biochemical properties of polymorphonuclear neutrophils from blood and peritoneal exudates of rabbits were compared. All enzymes measured showed almost identical activities in both types of cells, except for alkaline phosphodiesterase, the activity of which was seven times higher in peritoneal neutrophils. During phagocytosis, blood and peritoneal β-glucuronidases were released in very similar fashons. Lysozyme, one of the enzymes concerned with killing of bacteria, as well as β-glucuronidase, showed the same releasing pattern in both types of cells, but peroxidase was hardly released. Although superoxide anion generation in peritoneal neutrophils was two times higher than superoxide generation in blood neutrophils, phagocytic and bactericidal activities were almost the same in blood and peritoneal neutrophils. Blood neutrophils were more resistant to hypotonic lysis than were peritoneal neutrophils. These results show that there are no distinct differences in enzymatic and functional properties between blood and peritoneal polymorphonuclear neutrophils, except for alkaline phosphodiesterase activity, superoxide anion production, and osmotic fragility.

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The pathogenetic importance of oxygen intermediates (O.I.) which are highly reactive has been recognized recently. The present paper reports that human vascular damage may be mediated by granulocytes and this damage was inhibited by scavengers of O.I.. The cultured human endothelial cells (EC) derived from umbilical cord vein were used as in vitro model of the vessel. The hypoxanthine-xanthine-oxidase (HPX-XOD) system was used as the model of granulocytes which generate O.I.. EC damage was determined by trypan blue dye exclusion test or microcytotoxicity assay. EC damage in HPX-XOD system was marked and this damage was not inhibited by adding superoxide dismutase (SOD) alone. The catalase significantly inhibited EC damage, but incompletely. Simultaneous addition of both SOD and catalase in the system completely inhibited EC damage. Various concentrations of commercial H₂O₂ showed dose dependent EC damage. These results suggest that H₂O₂ is most highly destructive for EC and simultaneous additon of the scavengers was most effective for the inhibition of EC damage. The human granulocyte lysate which is rich in lysosomal enzymes, but does not generate O.I. showed EC damage in dose dependent fashion. The interaction of granulocyte and SLE serum leading to the release of O.I. into extracellular fluid may cause in vivo vascular damage. The granulocytes, on the other hand, may act to defend such oxygen toxicity in vivo (negative feed back system), releasing scavengers into plasma. The O.I. did neither affect Fe receptors on the granulocytes nor have any influence on the surface receptors of EC. The deposition of immune complexes in EC which had been reported experimentally and clinically seemed to be due to nonspecific micropinocytotic mechanism, not mediated by specific receptors.

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When jaw cysts are infected, polyunsaturated fatty acids, especially C 20: 4, constitution ratio decreases in the fluid of the cysts. The author considered that this change was because of the formation of lipoperoxides and prostaglandins from those fatty acids. The relation between the concentration of lipoperoxides and the infection of the cysts was studied. As a result, the infected cyst fluids showed high TBA value; when the infection was removed by irrigations in the cyst cavities, the value decreased and the cysts diminished in size. According to the author's investigations, it can be said that jaw cysts enlarge with the increase of lipoperoxides by the infection or the inflammation of jaw cysts.

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An increase of the urinary prostagrandin excretion from the patient with Bartter's syndrome has been reported in recent years far from the view point of the quantitative separation of PGs. Studies on profiling analysis of the urinary PGs using new HPLC separation method were executed in our laboratories. A combination system of the HPLC method using a reversed phase column with an extraction method by use of Kiesserguhr column was recommended for their profiling analysis as described in this paper.
During the untreated period with respect to a patient of this syndrome, 6-keto (6k) PGF_1α as a stable metabolite from PGI₂ and PGE₂ in the urine were found to increase markedly, when compared with the other PGs, while they were significantly reduced after the indomethacin therapy. However, those PGs less than several ng level were not detected by the present method. This simple and rapid HPLC method seems to be not only most useful for the profiling analysis of PGs in urine, but also applicable to those in other biological samples such as blood and tissues providing that the sensitivity in this separation method would be increased.

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Clinical, physiological and biochemical studies of PGE₁ were done, in a series of collagen diseas patients with skin ulcer, foot varix patient with skin ulcer as non-inflammatory skin ulcer control, and 2 diabetics without skin ulcer as no skin ulcer control. Intra-venous infusions of prostaglandin E₁ (PGE₁) was given continuously in the dose of 1 ng/kg/min for 72 hours. Blood samples were collected from cubital vein, before, during, right after and at 7 days after PGE₁ therapy. Platelet aggregations were studied by light transmittance (PRP: modified by Born's method, whole blood: modified by Tohjima's method) . Platelet iPG E (immunoreactive PG E like material) levels were assayed by radio-immunoassay. Essintial fatty acids compositions of plasma, pltelet and red cell were analyzed by gas chromatography.
Results were as follows:
(1) In all of cases, complete healing of skin ulcers was observed.
(2) In most cases, skin temperature increased during PGE₁ treatment.
(3) Platelet aggregation was increased during PGE₁ treatment than before one, and was higher in PRP than in whole blood, during PGE₁ treatment.
(4) The platelet basal iPGE levels were significantly decreased by PGE₁ (p<0.025) .
(5) The plasma and platelet linoleic acids levels were significantly higher than before PG E₁ treatment (plasma: p<0.05, platelet: p<0.025) .
(6) Thrombocytosis of one case of MRA was healed by the second PGE₁ treatment.
Conclusion:
The inflammatory skin ulcers in collagen diseases had healed completly by continued intravenous infusion of PGE₁. This effect might be brought by the supression of PG metabolism, especially in platelet.

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I measured serum protease inhibitors, α_l-antitrypsin, α_l-antichymotrypsin, α₂-macroglobulin, C 1 inactivator and inter-alpha-trypsin inhibitor by single radial immunodiffusion. I studied 15 normal controls and 52 patients with Behcet's disease.
In serum α_l-antitrypsin, α_l-antichymotrypsis, and inter-alpha-trypsin inhibitor showed an increase over normal controls, but the other protease inhibitors showed sligh increase.
A chromogenic sythetic tripeptide (Chrozym PK) was used as a substrate for kallikrein.
There was a tendency to low plasma kallikreinin in Behcet's disease. There was no correlation between plasma prekallikrein and CRP. The values of plasma kallikrein/ prekallikrein showed a sharp rise as plasma C 1 inactivator level was over about 40 mg/dl.

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In October, 1979, a 27 year-old male first had joint problems. Next month the skin lesions with itching were noted in the whole body. Steroid therapy (prednisolone 30 mg per day) provided minimal relief of the arthralgias. In January 1980, photophobia was complained with an obvious purulent discharge. He addmitted to our hospital in Febuary, 1980, because of slight fever, arthralgias of the extremities, keratodermia blennorrhagicum and circinate balanitis. He was treated with daily administration of phenylbutazone (200 mg) and azathioprine (100 mg) . Although conjunctivitis was disappeared and skin lesions have been improved, arthralgias have been still complained. Rentogenogram of the lumbosacral spine revealed the destructive change of the left sacroiliac joint unilaterally. HLA typing of lymphocytes showed HLA-B 27 in this patient. His synovial fluid revealed high levels of IgG, IgA and C 3. Large macropharges which had phagocytosed one or more polymorphonuclear leucocytes (Pekin's or Reiter's cell) were found in his synovial fluid. Those large macrophages and high levels of immunoglobulins and C 3 were also recognized frequently in Behcet's disease, therefore there may be a resemble inflammatory mechanism between Reiter's disease and Behçet's disease.

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A similar aberrancy of α₁-antitrypsis (α₁-AT) was detected in 8cases; their serum α₁-AT phenotypes (Pi) by acid starch gel electrophoresis seemed to be heterozygous of M and another one migrating more anodally. But these aberrant expression were suggested to be aquired. The common clinico-pathological abnormalities were in coagulation-fibrinolysis system. They were all diagnosed or strongly suspected to have disseminated intra-vascular coagulation.
568 sera with positive FDP (fibrin/fibrinogen degradation products) were tested for Pi. The same aberrancy of α₁-AT were detected in 23.1% of them. The relationship between FDP levels and frequancies of aberrancy is statistically highly significant.

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CCA [N- (2-carboxyphenyl) -4-chloroanthra-nilic acid] at 50-100 μg/ml markedly suppressed the PFC response when an optimal amount of antigen (SRBC) was present in the culture, while the drug did not suppress but enhanced the immune response in conditions where spleen cells were cultured with suboptimal amount of the antigen. On the other hand, the PFC response to DNP-Ficoll (a thymus-independent antigen) was not influenced by CCA. CCA, itself did not show mitogenic activity, but augmented the proliferative response caused by Con A. The proliferative response to a B-cell mitogen, LPS, however, was not affected by addition of 1-50 μg/ml CCA. Oral admiministration of CCA (10 mg/kg) increased the primary PFC response to TNP-HGG or SRBC in normal mice, but suppressed it in NZB/WF₁, The secondary antibody response to SRBC, however, was slightly sup-pressed by repeated administration of CCA between the first and second immunization. Moreover, the artificially enhanced antibody response by the injection of colchicine was tend to be restored to the normal level by CCA administration. Thus, the present experiments and other findings suggested that CCA might be a compound having im-munopharmacological profile as an immuno modulator sensitive to T-cell population. It would be expected that this drug can be applied not only to rheumatoid arthritis but also other immune diseases due to T-cell disfunctions.

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The efficacy of D-penicillamine in rheumatoid arthritis has now been clearly established, but the increasing use of this drug led many side effect. The common side effects are skin rashes, proteinuria, disturbance in taste and gastrointestinal upset.
We experienced two cases of rheumatoid arthritis with two major side effects of penicillamine, one episode of agranulocytosis and the other was myasthenia gravis. D-penicillamine was very effective in the both cases. The former was a 49-year-old woman who developed a precipitous fall in her granulocyte counts after about one month of penicillamine therapy at a daily dose of 300 mg. The latter was a 56-year-old man who developed a myasthenia like syndrome with diplopia and ptosis of left lid after about 5 months of penicillamine therapy at a daily dose of 200 mg.
Fortunately, the both patients were gradually improvement.
These side effects of penicillamine are very rare, and only two cases has been reparted in Japan, respectively. The physician must be familiar with these side effects and always pay attention to their appearrance during penicillamine therapy.

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The humoral immune response against a thymus-dependent antigen, sheep red blood cells (SRBC), was suppressed in mice in-jected with levamisole i.v. at a few days before challenge with the antigen. The suppressive effect of levamisole was modified by an antigen dose, a drug dose and a timing of drug administration. The suppression of plaque-forming cell (PFC) response was observed at an optimal antigen dose of 2×10⁸ SRBC while the augmentation was observed at a suboptimal antigen dose of 2×10⁷ SRBC, at the same dose of the drug. However, no suppressive effect on serum antibody level was found throughout the experiments. Levamisole also shifted the peak of IgM-PFC earlier and the peak of IgG-PFC later respectively than in normal responses. Transfer of spleen cells from levamisole-treated donors to untreated recipients resulted in antigen specific suppression in the recipient immune response.
These results suggest that, for the most part, an induction of antigen-specific suppressor cells in the spleen accounts for the suppression of humoral immunity by levamisole.

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We have previously presented that methyl-B₁₂ augments the din vitro IgM antibody response to SRBC and potentiates the proliferative response of lymphocytes to mitogens. In this paper the effects of several cobalamins and levamisole as well as methyl-B₁₂ in the in vitro antibody formation were evaluated, and it was shown that potentiating activity of methyl-B₁₂ was comparable to that of levamisole and highest in four cobalamins tested. In addition, the augmentative effect of methyl-B₁₂ in anti-SRBC PFC response was found not to be dependent on the increase of cell viability. Methyl-B₁₂ also potentiates the proliferative response of lymphocytes induced by 2μg/ml of ConA and these activated lymphocytes were found to show the suppressive effect to antibody response to SRBC. We suggest that suppressive effect of methyl-B₁₂ in experimental allergic encephalomyelitis reported by other investigators may be correlated to the potentiation in the generation of supnressor T cells.

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Site of analgesic action of tolmetin sodium was investigated in the acetic acid-induced writhing test in rats. There was a rough correlation between anti-writhing activity of tolmetin sodium and tolmetin contents in the peritoneal exudatewhen given orally. Anti-writhing ED₅₀ value of tolmetin sodium was 1.42 (0.82-2.91) and 92.0 (57.0-140) μg/kg when given i.p. and i.v., respectively, and the potency ratio of tolmetin sodium by i.v. to i.p. administration was 40.0 (18.5-80.2) . When tolmetin contents in the peritoneal exudate and the plasma were determined after i.p. or i.v. administration of tolmetin sodium with equipotent dose (5 μg/kg, i.p.; 200 μg/kg, i.v.), the content when given i.p. was nearly equal to and less than one-fortieth that when given i.v. in the exudate and the plasma, respectively. From these results, it was suggested that site of anti-writhing action of tolmetin sodium was in the peritoneum (periphery) in rats.

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Recently prodrugs are developed in various kinds of medicines. These prodruges are absorbed from gastrointestinal tract in inactive forms, and then converted to active forms mainly in the liver. In the field of non-steroid anti-inflammatory drugs, prodrugs are also being developed.
Using carrageenin edema method, we evaluated the four produrugs : Fenbufen, Sulindac, Suxybuzone and Acemethacine. By local Administratior, prodrugs show less antiinflammatory effects than active metabolites.
Then we discussed the appearance of side effects in the data of clinical trials carried out at many institutes. We found that the administration of prodrugs cause less side effects, especially the gastrointestinal ones, than that of active drugs.

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