Studying the effectiveness of irradiation on bone-marrow, the numbers of leukocyte, erythrocyte, hemoglobin of mice (25 normal mice and 35 mice treated by gamma irradiation with the dose of 600 rad/(100rad/day) (60 Co) showed that: Gamma irradiation reduced total of leucocytes, the number of different leucocytic (lymphocyte, granulocyte, mono and natural killer cells), the ratio of reticulocyte, number of mature erythrocyte and hemoglobin: Total of leucocytes (3,14 ± 1,58 in comparison with 13,45 ± 4,6); monocytes (0,05 ± 0,03 in comparison with 0,26 ± 0,13), lymphocytes (1,66 ± 0,36 in comparison with 6,34 ± 2,84). After gamma irradiation, the number of reticulocyte was 55%, mature erythrocyte was 73% and hemoglobin was 82%
Therapeutics ; Mice

An experimental of Vibrio vulnificus infection has been performed with the intravenous, subcutaneous and oral inoculation of Vibrio vulnificus into ICR mice. The results are as follows: 1) The LD50 of the intravenous, subcutaneous and oral inoculation of Vibrio vulnificus were 1.6x10(7) cells/ml, 4.0x10(7) cells/ml, and 2.5x10(9) cells/ml, respectively. 2) In the experimental group without treatment with CC14, the survival rates for intravenous inoculation were 100% (1/2 LD50), 39.1% (LD50), and 8.3% (2 LD50). The survival rates for subcutaneous inoculation groups were 100% (1/2 LD50), 46.9% (LD50), and 18.8% (2 LD50). And the survival rates for oral inoculation groups were 100% (1/2 LD50), 53.1% (LD50), and 43.8% (2 LD50). 3) In those treated with CC14 0.05 ml, the survival rates for intravenous inoculation groups were 43.8% (1/2 LD50), 29.1% (LD50), 0% (2 LD50). The survival rates for subcutaneous inoculation groups were 59.4% (1/2 LD50), 40.6% (LD50), and 9.4% (2 LD50). The survival rates for oral inoculation groups were 68.8% (1/2 LD50), 46.9% (LD50), and 18.8% (2 LD50). In those treated with CC14 0.1 ml, the survival rates for intravenous inoculation groups were 25.0% (1/2 LD50), 10.4% (LD50), and 0% (2 LD50). The survival rates for subcutaneous inoculation groups were 43.8% (1/2 LD50), 21.9% (LD50), 0% (2 LD50). The survival rates for oral inoculation groups were 50.0% (1/2 LD50), 37.5% (LD50), and 0% (2 LD50). 4) Liver, lungs, meninges and brain, kidneys, heart, gastrointestinal tract and spleen showed septic inflammatory findings. Their degree of inflammation were different according to the severity of hepatic damage and the inoculum size.
Mice ; Animals

30 Swiss mice inoculated by the i.p injection of 106 Sarcoma TG. 180 ascitic cells per each one. 10 mice were injected (i.p) only one with 35mg/kg dosage of complex at the 10th day after tumor inoculation. Another 10 mice were got ascites out of them and the injected drugs as above mentioned. Our experiments have got the following results: average life span of control mice is 14 days. Nonoperative treated mice: 22 days (increase 57.1%), one mouse was survived. Postoperative treated mice: 40 days (increase 185.7%), two mice were survived.
neoplasms ; Mice

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

No abstract available.
Animals ; Mice*

No abstract available.
Animals ; Mice*

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

BACKGROUND: Hypertrophy of podocytes is observed in type 2 diabetic patients. Cellular hypertrophy requires combined effects of various mitogen- induced entry into the cell cycle and subsequent cell cycle arrest at the G1/S interphase. This cell cycle arrest is mediated by various cyclin-dependent kinase inhibitors (CKIs). We investigated the effect of angiotensin II receptor blocker (ARB) treatment on podocyte hypertrophy and CKIs expression in cultured podocytes stimulated by long-term high glucose. METHODS: Immortalized mouse podocytes were cultured in media containing 5.6 mM normal glucose (NG), 30 mM high glucose (HG), or NG+angiotensin II (AII, 10(-7)M) for 7 days with or without ARB (L-158,809, 10(-6)M). Cellular hypertrophy was assessed by measurement of cellular protein/cell counts, and CKIs mRNA and protein expression were assessed by reverse-transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. RESULTS: Cellular hypertrophy was induced in podocytes exposed to HG or AII compared to NG cells and this HG-induced cellular hypertrophy was inhibited with ARB treatment by 70% (p<0.05). In addition, there were 1.5-fold and 2.0 fold increases in p27Kip1 mRNA and protein expression, respectively, in HG-stimulated podocytes compared to NG- treated cells (p<0.05). p27Kip1 mRNA and protein expression were also increased in cultured podocytes stimulated by AII by 156% and 199%, respectively (p<0.05). ARB treatment ameliorated HG-induced increase in p27Kip1 mRNA by 75% and protein expression by 70% (p<0.05). In contrast, there were no significant changes in p21Cip1 and p57Kip2 protein expression in cultured podocytes exposed to HG or AII. CONCLUSION: High glucose induced significant cellular hypertrophy and increased p27Kip1 mRNA and protein expression in cultured mouse podocytes, and these changes were effectively inhibited by ARB treatment.
Mice ; Animals