Interferons are protein that inhibit viral replication, slow cellular proliferation, influence immune response, etc. And significant progress have been made over the past five years. Recent clinical studies have defined clinical applications for both virus and malignant disease. Interferons show limited but definite clinical effects. However, interferons will not be used as solitary agents but rather as adjuvants in conjunction with other treatment modalities. Limited supplies of interferons have prevented adequate pharmacologic studies of optimal dose, route, and schedule. Now, enough interferons will be available so that interferon therapy will be less expensive than now. All of these factors will be important in design of future clinical trials. Taking all these factors into considerations, randamized phase III investigations will be necessary to define the ultimate role of interferons in the treatment of viral and malignant

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A simple and rapid microtechnique using 10 ul of whole blood was developed for studying a luminol-dependent chemiluminescence (CL) response. This assay have been thought to be unreliable because of poor reproducibility. In fact, the CL response of this assay was intrinsically affected by contamination of red blood cells or other cellular components and plasma constitutents. More importantly, there were several serious problems to get a reliable and reproducible measurement, because this CL respons by itself was varied hours by hours and day by day and different machines gave us a significantly different measurement to the CL response of the same blood sample. We carefully studied the effect of red blood cells and serum on CL response and found that if 10 ul of whole blood were used instead of 100 ul, this assay can be successfully and reliably used for evaluating the phagocytic function of peripheral blood phagocytes, especially by a simultaneous measurement of CL respons against PMA and OZ immediately after bleeding at a certain fixed time of a patient.
This simultaneous microtechnique of whole blood CL response will offer a very important knowledge of cell surface functions (FcR and C_3bR) of phagocytic cells, involved in the pathological events of various diseases.

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Serosal mast cells were purified by bovine serum albumin density gradient centrifugation and prelabeled with ³²PO₄. Incorporation of ³²PO₄ into PPI, diphosphoinostide (DPI) and triphosphoinositide (TPI), was determined by thin-layer chromatography on oxalic acid impregnated silica gel plates. ³²PO₄ incorporation into DPI in mast cell supernates as well as pellets was increased by concanavalin A. As a control for possible artifactual formation of DPI in the supernates, supernates from labeled stimulated or unstimulated cells were incubated with intact or broken membrane granules from unlabeled cells isolated from sonicated mast cells on a Percoll gradient. No obvious ³²PO₄ incorporation into PPI in the granules was seen. The question of how much of the labeled PPI is present in the granule fraction was considered. Studies in unstimulated mast cells labeled with ³²PO₄, including controls for artifactual incorporation of label during granule isolation, suggest that at least 40% of the total cellular radiolabel in PPI is in the granules. The results of these studies indicate that changes in PPI metabolism may be a intrinsic part of the biochemical mechanisms that control mediator release from mast cells.

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In order to study on the elastolysis and fibrinolysis in inflammatory process, the level of granulocyte elastase (ELP) was assayed in inflammatory diseases. The assay was carried out in children suffering from inflammatory diseases such as Kawasaki disease (MCLS), pneumonia, allergic purpura and others.
ELP was extracted from granulocytes by 2mol/l KSCN and measured by using the artificial substrate, Suc-Ala-Tyr-Leu-Val-pNA. The biological activity was expressed as amidolytic activity per cell. The ELP activity of normal newborn babies was 9.0±3.1 nU/cell and it increased by age, attaining to the level of 16.1±5.8 nU/cell of normal adults. In MCLS, pneumonia and aseptic meningitis, the level of ELP decreased during the 2nd and 3rd weeks of illness, and it returned to the normal range up to the 4th or 5th week. On the other hand, the level of ELP was not reduced in upper respiratory diseases in which the inflammation was moderate. In allergic purpura, the level of ELP increased and returned to the normal range after the 5th week. Although the causes of the decreased level of ELP in inflammatory diseases are not clear, it can be suspected that ELP is released from granulocytes and consumed during inflammatory process.

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Dexamethasone 21-palmitate (DP) is a main ingredient of Liposteroid which is an injectable anti-inflammatory preparation, in a form of fat emulsion. DP is considered to be a pro-drug. Many steroid esters used therapeutically are known to be rapidly hydrolyzed in vivo. However, the hydrolysis rate of steroid esters vary with chain length. It is, therefore, very important to know the hydrolysis rate of DP and tissue distribution of hydrolyzing enzymes in relation to its pharmacology, toxicity and metabolism in vivo. The hydrolyzing activities were found in serum, liver and macrophages of rat and also in liver and monocytes of human. The affinity of DP to the enzymes is very low and DP was found to be more slowly hydrolyzed in vitro. DP is likely to be hydrolyzed by non-specific carboxylesterases. These results suggest that DP is hydrolyzed gradually and continuously to dexamethasone and exhibits a more prolonged activity in vivo.

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Leukotriene B₄ (LTB₄) has chemotactic potency. Bovine lenses were incubated in Tyrode's solution and stimulated with calcium ionophore A 23187. LTB₄ released into the medium was identified by reversephase HPLC (RP-HPLC) . LTB₄ biosynthesized by bovine lenses was identified by its RPHPLC elution time and its chemotactic activity for neutrophils.
The result represents that LTB₄ synthesized in lenses and released into aq. humor under some condition could play an important role in inflammatory reaction.

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Recent study revealed that methyl B₁₂ had some immuno-modulating action. In this report we studied immuno-modulating action of methyl B₁₂ with serological change of synovial fluid to 10 RA patients. The effects were evaluated with pre and post administration of methyl B₁₂ on the Lansbury index, concentration of synovial immunoglobulin and complement.
Morning stiffness and joint index had significantly improved. In synovial fluid, uneffective group had more increased rate of immunoglobulin and effective group had a increased level of CH₅₀, C₃ but there were no statistical significance. In this study shows that methyl B₁₂ may have some immuno-modulanting action.

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Synovial fluids of patients with rheumatoid arthritis (RA) suppressed the proliferation of synovial cultured cells taken from patients with RA shown in reduction of the uptake of H³-thymidine into the cells.
Supernatants of cultured macrophages and polymorphonuclear leukocytes (PMN), but not lymphocytes in synovial fluids of RA patients could suppress the proliferation of the synovial cultured cells.
The culture supernatants of the macrophages in the synovial fluids treated with indomethacin could not suppress the proliferation of the synovial cultured cells and purified PGE₂ suppressed strongly the proliferation of the synovial cultured cells.
From these results, the suppressive effect of rheumatoid synovial fluids on the proliferation of synovial cultured cells might be to PGE₂ which was released from macrophages or PMN in the synovial fluids.

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Subsets of lymphocytes in synovial fluids of patients with rheumatoid arthritis (RASF-Ly) along with lymphocytes in peripheral bloods of normal subjects (Healthy-Ly) were studied.
Percent of OKT-3+cells (P<0.01), OKT-4+cells (P<0.01), OKT-8+cells (P<0.05) and OK-Ial+cells (P<0.01) of RASF-Ly were significantly higher than those of Healthy-Ly. Therefore, we examined the correlation these percentage of subsets. Positive correlation were observed in RASF-Ly between percentage of OKT-8+cells (P<0.01) or Leu-7+cells (p<0.01) and that of OK-Ial+cells and also between percentage of OKT-8+cells and that of Leu-7+cells of RAST-Ly (p<0.01) . Since OK-Ial represents a marker of activated T cells, both OKT-8+cells and Leu-7+cells in RASF-Ly seemed to be activated and Leu-7+cells seemed also to be immature. NK activity of RASF-Ly was significantly lower than that of Healthy-Ly (p<0.01), whereas ADCC activity of RASF-Ly was similar to that of Healthy-Ly.

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We administered liposteroid to rheumatoid patients and evaluated its usefulness. The liposteroid is dexamethasone palmitate incorporated into small lipid particles (lipid microspheres) and was developed for the purpose of maximizing therapeutic effects while minimizing side effects. In animal experiments liposteroid has shown a specific distribution in reticuloendothelial system and inflamed tissues, and a stronger anti-inflammatory activities than watersoluble dexamethasone.
In a double blind trial involving 36 RA patients a single dose of 2.5 mg liposteroid showed significantly (p<0.05) stronger and longer effects than 3.3 mg water-soluble dexamethasone. The incidence of side effects observed included headache, pain at the site of injection and itching.
Plasma cortisol levels recovered to pretreatment range within a week after injection of both materials. Long term observation of the patients using liposteroid revealed no lowering of plasma cortisol levels.
Based on above, we conclude that liposteroid is a useful drug for the treatment of rheumatoid arthritis.

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