Recently there is increasing tendency of the nosocomial infection, and Pseudomonas aeruginosa is one of the most important and common pathogens causing hospital opportunistic infections with rapid emergence of resistant strain especially in immunologically compromised patients. An experimental study for the effects of polyvalent Pseudomonas vaccine was performed in an animal model of Pseudomonas sepsis on a survival rates and histopathological points of view-using ICR inbred mice. The vaccine was prepared with heat killed whole cells of the 10 representative serotypes of Pseudomonas aeruginosa, which were isolated from the Department of Microbiology, College of Medicine, Kyung Hee University and Seoul National University, and they were devided into two polyvalent vaccine groups. The animal model of the Pseudomonas sepsis was deveoped by intravenous inoculation of Pseudomonas aeruginosa (serotype F, inoculum size 100 microliter, 109 cells/ml), immediately after cutaneous burns. The results were as follows. 1) The survival rate of the immune mice was 100% and that of non-immune mice was 60%. 2) The histologic findings of lung of the non-immune mice were severe congestion (18/18 mice), hemorrhage (18/18 mice), emphysematous change (18/18 mice), thrombosis (9/18 mice), infarction (9/18 mice) and inflammation (6/18 mice) and those of the immune mice were only congestion (6/20 mice) and focal emphysematous change (2/20) from the 3 day experimental group. 3) The histologic findings of the liver in the non-immune mice were severe congestion, Kupffer cell mobilization, focal necrosis, & portal inflammation in most of them, and from 7 day experimental group there were noted infiltrations of lagre histiocytic cells in sinusoids, and those in the immune mice were only reactive change of varying degree. 4) The histologic findings of the spleen in the non-immune mice were severe reactive hyperplasia in all and ischemic necrosis in about half of them, and those in the immune mice were only reactive change. 5) The histologic findings of the heart in the non-immune mice were severe congestion and inflammation in most and in the immune mice were only occasional nonspecific congestion. 6) The histologic findings of the kidney in the non-immune mice were severe congestion in all, interstitial inflammation, acute tubular necrosis and cortical necrosis in about half of them, and those in the immune mice were only mild congestion. With the above results, we can suspect there is a significant protective effects of the polyvalent pseudomonas vaccine on the pseudomonas sepsis in ICR mice.
Mice ; Animals

Viral myocarditis is considered an important cause of dilated cardiomyopathy. At preseent, two mechanisms are known to be involved in the pathogenesis of viral myocarditis and subse-quent cardiomyopathy: viral direct toxicity and immune mediated toxicity. Some authors have reported that IL-6 influences the immunologic mechanism and the virus-induced tissue damage in myocarditis. We injected encephalomyocarditis(EMC) virus to induce viral myocarditis in ICR mice. In order to study the lymphocyte subset and IL-6 expression to clarify the immune mechanism and to demonstrate the role of IL-6 in viral induced myocardial damage. The following results were obtained: 1) In virus inoculated mice, inflammation was severest at 10 days, and some serious complications developed, indicating a possible transition to dilated cardiomyopathy. 2) On analysis of the lymphocyte subset, CD4 cells were most prevalent at 5 days and CD8 cells were most prevalent at 10 and 20 days. 3) IL-6 was significantly increased and expression of IL-6 was constant, but its intensity was strongest at 5 days. In conclusion, IL-6, produced by inflammatory cells, fibroblasts, and endothelial cells, might play an important role in myocardial damage in experimentally induced EMC viral myocarditis by its direct cytotoxicity or cytokine mediated activation of cytotoxic cells.
Mice ; Animals

No abstract available.
Animals ; Mice*

Transglutaminase(TGase) is a calcium dependent enzyme that catalyse and acyl transfer reaction forming epsilon-(gamma-glutamyl)-lysine cross linkage. the major known effect of TGase is its important role in the programmed cell death manifested in the granular layer of the skin and acidophilic bodies in the viral hepatitis and neoplastic processes. The enzyme activity, immunohistochemical reaction using polyclonal antibodies against cytosolic TGase C, light and electron microscopic studies and TdT staining of the transplanted fibrosarcoma cells in C3H mouse with radiation therapy were done. The presence of TGase was detected immunohistochemically by avidin-biotin peroxidase complex (ABC) method Apoptosis were significantly induced after irradiation dependent with time factors and irradiation doses, resulted in marked and confluent tumor cell loss. Highest activity of the cytosolic form of TGase was noted at 24 hours and decrease after then while membrane bounded form of the TGase showed no significant changes. Immunohistochemical staining revealed strong positive reaction in the sarcoma cells in diffuse fasion and around the necrotic foci in the cytoplasm. Terminal dideoxynucleotidyl transferase(TdT) staining revealed increasing numbers of apotptic cells from two hours after irradiation. In the mechanism of decreasing tumor size and cell death in radiation therapy, apoptosis plays an important role and during that process transglutaminse might do some irreversible cross-linking effects of cytoplasmic proteins causing cell death in part.
Mice ; Animals

OBJECTIVE: Indoleamine 2, 3-dioxygenase (IDO), an immuno suppression enzyme, is one of the initial and rate-limiting enzymes involved in the catabolism of the essential amino acid tryptophan. IDO inhibits T cell proliferation, induces T cell apoptosis, and plays a fundamental role in autoimmunity and allergy. We investigated which subtype of dendritic cells (DCs) is involved in IDO expression and the generation of regulatory T cells during the induction of oral tolerance in type II collagen-induced arthritis (CIA). METHODS: Type II Collagen was fed to DBA/1J mice before immunization. Changes in DC subtypes and induction of regulatory T cell in orally tolerized CIA mice were analyzed. Whether the effect of DC subtype was modulated by the IDO expression, was determined by flow cytometry (FACs) and confocal microscopy. RESULTS: IDO expression of CD11c+ DCs was higher in orally tolerized CIA mice than in non-tolerized CIA mice. CD11b+ DCs of the CD11c +DCs, subtype was higher in the induction of in IDO expression. Our data suggest that these IDO expressing DCs of oral tolerized mice suppressed type II collagen-specific T cell proliferation and favored the differentiation of naive CD4+ T cells into regulatory T cells. Especially, CD11c+CD11b+ DCs expressed IDO, which is known to be associated with regulatory T cell induction. CONCLUSION: We observed that oral tolerance induced the increase in IDO-expressing CD11c+CD11b+ DCs, which appeared to induce regulatory T cells. IDO-expressing CD11c+CD11b+ DCs are involved in oral tolerance, which may provide a new therapeutic approach for the treatment of rheumatoid arthritis.
Mice ; Animals

Bone resorption involves sequential stages of osteoclast precursor migration and differen-tiation of osteoclast precursors into multinucleated osteoclasts. Stromal cell derived factor (SDF)-1 is a chemotactic factor for osteoclast precursor migration. Matrix metalloproteinase (MMP)-9 is involved in migration of osteoclast precursors and activation of interleukin(IL)-1beta. Alveolar bone destruction is a characteristic feature of periodontal disease. Treponema lecithinolyticum is a oral spirochete isolated from the periodontal lesions. The effect of lipopolysaccharide(LPS) from T. lecithinolyticum on expression of SDF-1 and MMP-9 was examined in cocultures of bone marrow cells and osteblasts derived from mouse calvariae. T. lecithinolyticum LPS increased expression of MMP-9 in the coculture. Polymyxin B, an inhibitor of LPS, abolished the increase of MMP-9 mRNA expression by LPS. LPS did not increase the expression of SDF-1, IL-1beta and tumor necrosis factor(TNF)-alpha mRNA in cocultures. Prostaglandin E2(PGE2) up-regulated the expression of MMP-9 and NS398, an inhibitor of PGE2 synthesis, down-regulated the induction of MMP-9 expression by T. lecithinolyticm LPS. These results suggest that T. lecithinolytium LPS increases MMP-9 expression in bone cells via PGE2 and that the induction of MMP-9 expression by T. lecithinolyticum LPS is involved in alveolar bone destruction of periodontitis patients by the increase of osteoclast precursor migration and the activation of bone resorption-inducing cytokine.
Mice ; Animals

Bone resorption involves sequential stages of osteoclast precursor migration and differen-tiation of osteoclast precursors into multinucleated osteoclasts. Stromal cell derived factor (SDF)-1 is a chemotactic factor for osteoclast precursor migration. Matrix metalloproteinase (MMP)-9 is involved in migration of osteoclast precursors and activation of interleukin(IL)-1beta. Alveolar bone destruction is a characteristic feature of periodontal disease. Treponema lecithinolyticum is a oral spirochete isolated from the periodontal lesions. The effect of lipopolysaccharide(LPS) from T. lecithinolyticum on expression of SDF-1 and MMP-9 was examined in cocultures of bone marrow cells and osteblasts derived from mouse calvariae. T. lecithinolyticum LPS increased expression of MMP-9 in the coculture. Polymyxin B, an inhibitor of LPS, abolished the increase of MMP-9 mRNA expression by LPS. LPS did not increase the expression of SDF-1, IL-1beta and tumor necrosis factor(TNF)-alpha mRNA in cocultures. Prostaglandin E2(PGE2) up-regulated the expression of MMP-9 and NS398, an inhibitor of PGE2 synthesis, down-regulated the induction of MMP-9 expression by T. lecithinolyticm LPS. These results suggest that T. lecithinolytium LPS increases MMP-9 expression in bone cells via PGE2 and that the induction of MMP-9 expression by T. lecithinolyticum LPS is involved in alveolar bone destruction of periodontitis patients by the increase of osteoclast precursor migration and the activation of bone resorption-inducing cytokine.
Mice ; Animals

Dendritic cell (DC)s are protessional antigen presenting cells and they have been used for antitumor immunotherapy or cell vaccines. However therapy using DC is restricted because the number of DC available from tissue is very low. Flt3 ligand (FL) has been known as a hematopoietic growth factor that increases proliferation of hematopoietic stem cells and progenitor cells, and recently it showed inducibility of dendritic cell (DC)s and signiticant antitumor effects in vivo. Thus FL will be frequently used for DC induction and antitumor immunotherapy in future. Here we constructed FL plasmid and studied its in vivo effect. FL plasmids were made by cloning of partial FL cDNA into pcDNA3 plasmid, and gene expression and protein producibility of FL plasmid were confirmed in Renca cells transfected with FL plasmid. Mice were injected with FL plasmid (100ug/mouse) three times and 20 days later mouse spleens were harvested for staining and RT-PCR. There were lots of blastogenic cells in the spleen of mice treated with FL plasmid. FL plasmid also induced DEC205, IL-12 and GM-CSF gene expression in mouse splenocyte. All these data suggest FL plasmid may be used for induction of DC and antitumor therapy as DNA adjuvant.
Mice ; Animals

Regarding numerical changes of leukocytes involved in immune defects following thermal injury, a lot of controversial results have been reported. In this study, the changes of leukocyte number and distribution were examined and compared in spleen and lymph nodes of thermally injured mice. Mice (Balb/c) were anesthetized by intraqeritoneal injection of 2,2,2-tribromoethanol and thermally injured by immersion of hair-removed dorsal skin (15% total body surface) in a boiling water bath (96`C) for 7 seconds. Both of lymph node cell (LNC) and spleen cell (SPC) numbers decreased significantly at day 2 of injury and thereafter rebounded, but in a distinct pattern; 1) LNC numer returned to over normal level at day 6 and normalized again, whereas SPC number increased gradually over normal level and sustained until day 24 of injury. 2) Such increase of LNC and SPC number coincided with higher proportion of PMN and relative decline of lymphacytes, particularly CD3 T cells rather than slg' B cells, but such alteration was more significant in spleen. The changes of peripheral blood leukocyte (PBL) number was comparable to those of SPC. These data suggest that the cause of immune modulation in thermally injured mice acts systemically. In addition, it is noteworthy that reduction of lymphocyte and CD3 T cell proportions was due to relative increase of PMN number, not the decrease of absolute number of lymphocytes. Spontaneous recovery of injured mice in this study also implicates that increase of PMN number may be responsible for recovery from injury without infection. Finally, the CD4'/CD8' ratio of injured mice was lower only at day 2 ot injury, but not significantly, than that of control group. It is likely that contribution of Th/Ts ratio to immune defect after thermal injury should be determined together with other factors, such as injured body surface % and severity of injury.
Mice ; Animals

We have used BALB/c mice as an animal model for the study of anaphylactic hypersensitivity to the house dust mite. For the sensitization, BALB/c mice were injected with a single dose of extracts of Oermatophagoides farinae (D. pa) or Dermatophagoides pteronyssinus (D. pt) mixed with adjuvants (aluminum hydroxide and Bordetella pertussis) intraperitonealy. On days of 15, 30, and 60 after the sensitization, the mice received a challenge dose of the same allergen intravenously to induce anaphylactic shock. The hypersensitivity reactions were scored by anaphylactic shock. And various immunological parameters, including cytokines and immunoglobulin isotypes, were studied in relation with the shock. A high level of anaphylactic shock was produced in the mice by both of the allergens, D, fa and D, pt, at 15 and 30 days after sensitization. In vitro Ag specific proliferative reponses of spleen cells from D. pt treated mice (D. pt mice) was six times higher than those from O. fa treated mice (O. fa mice). Regardless the differences in antigens, the production of IFN-r by spleen cells from D. pt mice or O. fa mice was equally high at 15 days after sensitization. However, the ability to produce IFN-r by the spleen cells from D, pt mice was three times higher compared to that from D. fa mice. The production of IL-4 by the spleen cells was enhanced slightly but not significant in both groups. In studies of the allergen-specific immunoglobulin isotypes in the sera of the mice, the level of IgE in both groups was enhanced slightly but not significant. In contrast, the level of IgG subtypes were increased in both groups. When the levels of IgG were compared by subtypes, the level of IgG1 increased significantly on day 15 when the anaphylactic shock score was maximized in both groups. Increase in IgG2a level at the day was not significant, instead, asignificant increase in IgG2 levels was observed on day 60 after sensitization when the anaphylaxis was almost discontinued. Although a higher level of IgG3 was examined on day 15 and 30 in D. pt mice and on day 60 in D, fa mice, anaphylaxis was not appeared to be associated with the levels of IgG3 in this study. The IgG1, rather than IgE, was assumed to the major factor involved in the anaphylactic response observed in this experiment. In conclusion, BALB/c mice would be an animal model for the study of anaphylactic hypersensitivity to D. fa or D, pt., which might be an essential tool for the future development of immuno-therapeutic agents.
Mice ; Animals