The increased susceptibility in patients of chronic renal failure to infection has been reported to be attributed to defects in granulocyte and lymphocyte function and proliferative activity of hematopoietic cells. The definite cause of the frequent infection in uremic patients, however, is still controversial. The effect of uremic plasma on the aspect of the hematopoietic cells has been scarcely been studied. In the present study, mouse bone marrow was cultured with uremic plasma, to evaluate the effect of uremic plasma on the proliferative activity and morphological features of CFU-GM. The results obtained were as follows. 1) The number of colonies in group co-cultured with uremic plasma was more reduced than that of normal plasma group. 2) There was no difference between the group cultured with predialytic uremic plasma and that of postdialytic plasma in number of colonies, macroclusters and microclusters. 3) The forms of colony were granulocytic and monocytic forms at 5 day of culture. Electron microscopically, granulocytes disclosed electron dense azurophilic granules and electrolucent specific granules in the cytoplasm, and monocyte showed numerous vesicles and vacuoles in the cytoplasm which had finger-like projections. 4) The molecular weight of inhibitory factor in the uremic plasma was supposed to be less than 50,000 daltons.
Mice ; Animals

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

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

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

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

Alveolar bone destruction is a characteristic of periodontal disease. Treponema denticola are found in significantly increased numbers in the sites affected with periodontal disease. In order to clarify the role of T. denticola in destruction of alveolar bone in periodontal disease, this study was undertaken to determine the effect of sonicated extract of T. denticola on osteoclast differentiation in co-culture system of mouse bone marrow cells and calvaria cells. The ability of osteoclast formation was estimated by counting the number of tartrate resistant acid phosphatase(TRAP) positive cells. Sonicated extract of this bacteria stimulated osteoclast formation in a dose dependent manner(p<0.05). Indomathacin, an inhibitor of prostaglandin synthesis, decreased osteoclast formation induced by sonicated extract of this bacteria(p<0.05). Extract-induced osteoclast formation was decreased, when sonicated extract of bacteria was heated(p<0.05). These findings suggest that T. denticola induces osteoclast differentiation, and protein component of this bacteria and PGE2 may play an important role in this process.
Mice ; Animals

The mechanism of cytolysis by complement attack of nucleated cells(NC) is of special interest in comparison to that of red blood cells. It is known that NC death by membrane attack comples, C5b-9, is caused by many factors, i.e., efficiency of complex assembly, activation of intrinsic metabolic pathway by signal transduction, cytotoxic effect of the channel itself and natural repair ability. These factors suggest that colloid osmotic lysis, known in red blood cells, does not fully explain the complement-mediated cell death of NC. In this study, the authors investigated correlation between biochemical and morphological changes to prove "Ca2+-mediated metabolic death"8~13) representing a mechanism of NC death caused by C5b-9 attack. The L1210 cells, mouse leukemic cell line carrying small complement channel(TAC5b-91) were used in the experiments. The amounts of intracellular adenine nucleotides to extracellular Ca2+, ouabain, KC1 and dextran were analyzed by bioluminescence method using luminometer. Cell viability was checked by 0.4% trypan blue dye and LDH release. Morphological observation of TAC5b-91 was done by immunocytochemical staining and electron microscope. The results were as follows: 1) The release of ATP, ADP and AMP followed by cell death was rapid and progressive along the incubation time at 37 degrees C and it was accelerated in 1.5 mM of [Ca2+]0. 2) There was no evidence of ATP repairment in the TAC5b-91. 3) Extracellular KC1(150 mM), dextran(0.66 mM) and ATP supplement(0.2 microM) could not effectively inhibit ATP depletion and cell death. Ouabain(27 and 100 microM) enhanced cell death and could not completely prevent ATP loss. 4) Most of the mitochondria showed swelling, loss of cristae and Ca2+ deposit in matrix in the electron microscopic observation. Rapid, sustained and irreversible depletion of adenine nucleotides was due to Ca2+ deposit with destruction of mitochondria and also the leakage through transmembrane channels. Moreover this energy depletion was accelerated by high extracellular Ca2+ concentration. These results indicate that Ca2+-mediated, energy exhaustion is one of the mechanisms of the metabolic cell death by C5b-9 attack of NC.
Mice ; Animals

No abstract available.
Animals ; 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

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