Search

The anti-inflammatory effect of lanoconazole (LCZ) was investigated in vivo and in vitro. The effect of LCZ was evaluated on the inflammatory reactions elicited by intradermal injection of ovalbumin to ovalbumin-immunized rabbits, as an Arthus phenomenon. A one or two % cream preparation of LCZ was topically applied on the lesion daily after challenging injection until the inflamation had diminished. By macroscopic observation and measuring the diameter of edema, erythema, hemorrhage and necrosis, the effects of LCZ on the reactions were compared with the reactions of the sites administered withcream vehicle as reference agent. Two % LCZ showed an anti-hemorrhagic effect. The in vitro effect of LCZ on production and functions of an inflammatory cytokine, TNF was also examined. LCZ suppressed the production of TNF by murine peritoneal macrophages at 20 micro g/ml and the adhesion of neutrophils at 100 micro g/ml. Moreover, LCZ significantly suppressed the growth inhibitory activity of TNF against L929 fibroblasts at 0.5 micro g/ml. A very low concentration of LCZ might protect the fibroblasts from immunological cytotoxicity in vivo. These findings suggest that LCZ has a suppressive activity to inflammatory responses and this suppressive action may be due to its protective activity to cells like fibroblasts.

The present studies were undertaken to clarify the difference of the immunogenicity evoked in rabbits immunized with a low-molecular chemical antigen (TNBS) alone, by four administration routes. Rabbits were divided into four experimental groups: TNBS sc group, i.v. group, im group and ip group (3 times/week, 10 times in total), and immune responses to TNBS were estimated by the assay systems of humoral immunoreactions (HA.PCA.Arthus) and cellular immunoreaction (DTH). The highest evoking of the immunogenicity to TNBS was noted in the sc group in the case of humoral immune response, whereas the lowest evoking in the i.v. group. The intermediate degree of the immunogenicity was elicited in the im and ip groups. On the other hand, only the sc group displayed positive cellular immune response (DTH), while such immune response was never recognized in the i.v., im and ip groups. These results were indicated that the immunogenicity to the low-molecular antigen (TNBS) was evoked exceedingly in rabbits by sc sensitization route from view of humoral and cellular immune responses.

The present study was carried out using a low-molecular antigen to clarify which of the two IgG subclasses (IgG1 and IgG2) of guinea pigs is responsible for the immunological assay systems in ordinary antigenicity tests for the development of a novel drug, and to what extent such IgG subclass antibodies contribute to the assay systems in addition to IgM antibody. Guinea pigs were immunized with TNBS plus FCA (3 mg/body), at intervals of 10 days, 3 times in total. The anti-TNBS serum was fractionated into peak I (IgG2), peak II (IgG1) and peak IV (IgM) by DEAE-cellulose column chromatography, and the immunological activities as well as the functions of the above three peaks were estimated by HA, homologous PCA reaction, PSANA and P-Arthus. The IgM and two IgG peaks possessed immunological activities on HA and P-Arthus, and the following degree of activities was shown in both assay systems: peak II (IgG1) greater than peak IV (IgM) greater than peak I (IgG2). In the homologous PCA reaction and PSANA, immunological activities were seen only in peak II (IgG1). It is confirmed that the IgG1 subclass is a homocytotropic antibody involved in the PCA and PSANA assay systems in the case of low-molecular immunogens such as TNBS.

The 48-hour passive cutaneous anaphylaxis (PCA) and passive anaphylactic bronchoconstriction in rats induced by an IgE-like antibody against DNP-Ascaris were inhibited by intravenous treatment with traxanox sodium in a dose dependent manner. In both experiments, traxanox sodium was more potent than disodium cromoglycate (DSCG), especially as an inhibitor of bronchial anaphylaxis. In the PCA test of rats using a double sensitization technique according to the Orr's method, traxanox sodium was demonstrated not to inhibit antigen-antibody combination, but to inhibit the release of chemical mediators at a stage following antigen-antibody combination. Traxanox sodium inhibited the complement dependent immune hemolysis, but not the hypotonic hemolysis in vitro. However it failed to inhibit the Forssman anaphylaxis in the guinea pig in vivo. Traxanox sodium (50-250 mg/kg p.o.) showed an inhibitory effect on the direct passive Arthus reaction (DPAR) of the rats. Furthermore, it delayed the onset of the hyperacute form of experimental allergic encephalomyelitis (EAE) and reduced mortality in the rats. DSCG was less effective on DPAR and EAE. In conclusion, traxanox sodium is considered to have a wider spectrum of anti-allergic activity than DSCG since it has a suppressive effect not only on the type I allergic reaction, but also on the type III and IV allergic reactions.

Experimental arthritis was induced in rats by the use of horse ferritin as an antigen. The pathological findings were studied and the mechanisms of inflammation were analyzed. Arthritis was induced through various mode of actions: 1. Arthus reaction (antigen-induced arthritis). 2. Passive Arthus reaction. 3. Reverse passive Arthus reaction. 4. Arthritis induced by intra-articular injection of immune complex, by either single shot or repeated injections. 5. Arthritis induced by anamnestic reaction. Arthritis induced by repeated injections of immune complex or that induced by anamnestic reaction exhibited mild chronic synovitis characterized by round cell infiltration without detectable acute phase of inflammation. Arthritis induced through other immunological mechanisms revealed a sequential course of acute synovitis characterized by accumulation of polymorphonuclear leucocytes (PMN) followed by gradual disappearance of PMN and alternative increase of round cells or plasma cells and proliferation of synovial lining cells. All types of arthritis subsided with mild fibrosis of the synovial tissue without lasting over a significantly long time. Although the successful production of a long-lasting chronic arthritis has been reported in rabbits, it was impossible to induce such long standing arthritis through this experiment. The chronicity of the arthritis induced through immunological mechanism may not simply be due to the long-standing retention of immune complex within the involved joint but also due to other factors such as the species of the test animals, the nature of antigen used and local factors produced through acute joint inflammation.

The pathological examination of the rabbit knee joint with antigen-induced arthritis produced by heat-killed Streptococcus faecalis (Str. faec.) as antigen was carried out. Macroscopically, there were findings of acute inflammation about five hours after the injection. Histopathologically, very remarkable acute exudative inflammation was seen 48 hours later. This supported the picture of Arthus reaction. The Arthus reaction disappeared with time, and this supported the view of delayed-type hypersensitivity three weeks later. After that, an obvious chronic inflammation was admitted in 10 weeks. This resembled the histopathological feature of rheumatoid arthritis (RA) when these findings are summarized. It was suggested that arthritis produced by Str. faec. progresses from an acute condition to a one with time. As mentioned above, it thought that Arthus reaction of both immediate hypersensitivity and delayed-type hypersensitivity are necessary in the occurrence of this arthritis.