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

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

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

PURPOSE: Oral mucositis induced by radiotherapy to the head and neck area, is a common acute complication and is considered as the most severe symptom for cancer patients in the early stages of treatment. This study was proposed to establish the oral mucositis mouse model induced by a single dose of radiation for the facility of testing therapeutic candidates which can be used for the oral mucositis treatments. MATERIALS AND METHODS: Fifty-five BALB/c mice were divided into four groups: control, 16 Gy, 18 Gy, and 20 Gy. Oral mucositis was induced by a single dose of radiation to the head and neck using 6 MV x-Ray from linear accelerator. After irradiation, body weight and physical abnormalities were checked daily. Tongue tissues from all groups were taken on days 1, 2, 3, 5, 7, 9, and 14, respectively and H&E staining was conducted to examine morphological changes. RESULTS: Body weight dramatically decreased after day 5 in all irradiated mice. In the 16 Gy treatment group, body weight was recovered on day 14. The histology data showed that the thickness of the epithelial cell layer was decreased by the accumulated time after radiation treatment, up to day 9. Severe ulceration was revealed on day 9. CONCLUSION: A single dose of 16 Gy is sufficient dose to induce oral mucositis in Balb/C mice. Significant changes were observed in the Balb/C mice on days 7 and 9 after radiation. It is suggested that this mouse model might be a useful standard tool for studying oral mucositis induced by radiation.
Mice ; Animals

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

PURPOSE: To investigate synergistic effect of local cryoablation with systemic immunotherapy, the tumor control ability and immunologic responses of combining these two modalities was compared with that of cryoablation, surgical excision, and immunotherapy only group in a tumor re-challenge model. MATERIALS AND METHODS: Preliminary experiments were performed in two stages. The first stage consisted of 36 Balb/c mice with Renca bearing tumors imbedded in the right thigh, and was treated with interleukin-2 (IL-2) and interferon-alpha(IFN-alpha) to evaluate the efficacy of immunotherapy and to determine the adequate dosage. The second stage was performed on 10 mice, to evaluate histological changes and efficacy after cryoablation. The main experiment was performed on 48 mice, divided into 6 groups of control with tumor implantation, excision of tumor, excision combined with immunotherapy, cryoablation of tumor, cryoablation with immunotherapy and control without tumor. After treatment, tumor re-challenge was performed with Renca cell, then the growth pattern was evaluated with physical measurements, and immune response was investigated with fluorescent activated cell sorter and cytotoxicity assay. RESULTS: Preliminary studies on immunologic efficacy revealed that IL-2 and IFN-alpha have a dose dependent inhibition of tumor growth. The main experiment evaluating the efficacy of combination treatment revealed that cryoablation with immunotherapy proved to be most effective in terms of tumor recurrence and tumor growth inhibition, yet the difference was not statistically significant from monotherapy with cryoablation. However, cytotoxicity was significantly increased cryoablation with immunotherapy compared with other groups. CONCLUSIONS: Cryoablation on tumor re-challenge mice model showed advantages with immunotherapy most prominently in cytotoxicity.
Mice ; Animals

The interaction of radiation and 5-luorouracil (5-U) on mouse jejunal crypt cells was studied using the microcolony survival assay. 150mg/kg of 5-U was injected intraperitoneally 15 minutes before irradiation and 6 hours after irradiation. Jejunal crypt cells of mouse survived more when 5-U was given 15 minutes before irradiation than giving it 6 hours after irradiation. The mean lethal doses (Do) of each of irradiation alone group, 5-U injection group of 15 minutes preceding irradiation, and 5-U injection group of 6 hours post irradiation were. 135, 135, and 114 rad respectively. The dose effect factor (DEF) of each of 5-U injection groups of 15 minutes preceding irradiation and of 6 hours post irradiation were 1.13 and 1.27.
Animals ; Mice*