• Title/Summary/Keyword: mouse spleen

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Isolation and Characterization of Single-Chain Fv Against Ductal Cells

  • Lee Myung-Hoon;Ryu Hye-Myung;Kim Sun-Zoo;Park Ji-Young;Uhm Ji-Hyun;Park Tae-In
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.211-217
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    • 2004
  • For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

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Immunostimulatory Activity of Hibiscus syriacus L. Leaves in Mouse Macrophages, RAW264.7 cells, and Immunosuppressed Mice

  • Na Gyeong Geum;Ju Hyeong Yu;So Jung Park;Min Yeong Choi;Jae Won Lee;Gwang Hun Park;Hae-Yun Kwon;Jin Boo Jeong
    • Korean Journal of Plant Resources
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    • v.35 no.6
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    • pp.697-703
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    • 2022
  • Under the COVID-19 pandemic, interest in immune enhancement is increasing. Although the immune-enhancing activity of plants of the genus Hibiscus has been reported, there is no study on the immune-enhancing activity of H. syriacus. Thus, in this study, we investigated the immune-enhancing activity of Hibiscus syriacus leaves (HSL) in mouse macrophages, RAW264.7 cells, and immunosuppressed mice. HSL increased the production of immunostimulatory factors such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) and activated the phagocytosis in RAW264.7 cells. The HSL-mediated production of immunostimulatory factors was dependent on toll-like receptor 4 (TLR4), p38, and c-Jun N-terminal kinase (JNK) in RAW264.7 cells. In the immunosuppressed mouse model, HSL increased the spleen index, the levels of the cytokines, and the numbers of lymphocytes, neutrophils, and monocytes. Taken together, HSL may be considered to have immune-enhancing activity and be expected to be used as a potential immune-enhancing agent.

Cytotoxicity of resident and Iymphokine-activated mouse peritoneal macrophage against yrichomonas vaginalis (질트리코모나스(Trichomonas waginazis)에 대한 마우스 복강 대식세포의 세포독성)

  • Yu, Jae-Suk;An, Myeong-Hui;Min, Deuk-Yeong
    • Parasites, Hosts and Diseases
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    • v.28 no.2
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    • pp.85-90
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    • 1990
  • This study was aimed to observe the direct and Iymphokine-activated cell mediated cytotoxic effects against Trichomenas waginalis by mouse peritoneal macrophages. Cytotoxicity was measured as release of 3H-thymidine from prelabeled protozoa, and tested in U-bottom microtiter plates. A 0.1 ml suspension of labeled protozoa (2{\times}10^5/ml$) was placed in each well, followed by 0.1 ml of a suspension containing increasing numbers of peritoneal cells. After a 24 hr incubation at $37^{\circ}C$, 0.1ml of the supernatant was collected and counted in liquid scintillation counter. Mouse peritoneal macrophages had appreciable level of spontaneous cytotoxicity against T. maginalis at the effector to target cell ratios from 5 : 1 to 50 : 1, Treatment of macrophages with Iymphokine, produced by PHA-stimulated spleen cells, increased the cytotoxicity in comparison with resident macrophages against T. vaginalis. The degree of macrophage activation for the killing was not dependent upon the Iymphokine concentration. Peritoneal cells adherent to plastic displayed significant levels of cytotoxicity against T. vaginalis. This study indicates that mouse peritoneal macrophages are spontaneously cytotoxic for T. waginalis and Iymphokine increases the cytotoxicity by activating macrophages to kill T. vaginalis.

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Survey on environmental condition and health state of laboratory mouse in conventional facility (일반시설에서 사육되는 마우스의 품질향상을 위한 기초조사 연구)

  • Lee, Heungshik S.;Seung, Rho Hyun;Kim, Kyungjin;Kim, Chul-kyu
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.611-625
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    • 2000
  • For the improvement of quality control of laboratory mouse, we investigated the environmental condition, histopathological findings and serological test using ELISA to mouse hepatitis virus(MHV), Mycoplasma pulmonis(MP), Clostridium piliforme(TZ) and Sendai virus (HVJ) of ICR, C57BL/6, CBA and C3H/He mice that were supplied from conventional laboratory animal facility. 1. The ammonia concentration of facility was below the recommended concentration, 15ppm, by the KNIH, and the room temperature($21{\sim}23^{\circ}C$) and relative humidity(40~60%) was optimum range recommend by the Ministry of Health and Welfare, respectively. 2. The incidence rate of inapparent disease was 86.6% and the major findings in the liver were vacuolar degeneration with nucleic pleomorphism. The lung was shown the thickening of alveolar wall and interstitial pneumonia with congestion. The kidney and spleen were observed the mild congestion and extramedullary hematopoiesis, respectively. 3. The positive reaction rates against MHV and MP in serological test was 97.9% and 37.5%, respectively but HVJ and TZ were negative. These results suggest that laboratory mice could be infected with MHV and MP under conventional environments. Therefore we recommend to select thoroughly inapparent infected mice and to convert conventional system into SPF facility as soon as possible.

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Antitumor Activity of the Korean Mistletoe Lectin is Attributed to Activation of Macrophages and NK Cells

  • Yoon, Tae-Joon;Yoo, Yung-Choon;Kang, Tae-Bong;Song, Seong-Kyu;Lee, Kyung-Bok;Her, Erk;Song, Kyung-Sik;Kim, Jong-Bae
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.861-867
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    • 2003
  • Inhibitory effect of the lectins (KML-C) isolated from Korean mistletoe (KM; Viscum album coloratum) on tumor metastases produced by murine tumor cells (B16-BL6 melanoma, colon 26M3.1 carcinoma and L5178Y-ML25 lymphoma cells) was investigated in syngeneic mice. An intravenous (i.v.) administration of KML-C (20-50 ng/mouse) 2 days before tumor inoculation significantly inhibited lung metastases of both B16-BL6 and colon 26-M3.1 cells. The prophylactic effect of 50 ng/mouse of KML-C on lung metastasis was almost the same with that of 100 $\mu$ g/mouse of KM. Treatment with KML-C 1 day after tumor inoculation induced a significant inhibition of not only the experimental lung metastasis induced by B16-BL6 and colon 26M3.1 cells but also the liver and spleen metastasis of L5178Y-ML25 cells. Furthermore, multiple administration of KML-C given at 3 day-intervals after tumor inoculation led to a significant reduction of lung metastasis and suppression of the growth of B16-BL6 melanoma cells in a spontaneous metastasis model. In an assay for natural killer (NK) cell activity. i.v. administration of KML-C (50 ng/mouse) significantly augmented NK cytotoxicity against Yac-1 tumor cells 2 days after KML-C treatment. In addition, treatment with KML-C (50 ng/mouse) induced tumoricidal activity of peritoneal macrophages against B16-BL6 and 3LL cells. These results suggest that KML-C has an immunomodulating activity to enhance the host defense system against tumors, and that its prophylactic and therapeutic effect on tumor metastasis is associated with the activation of NK cells and macrophages.

C1qa deficiency in mice increases susceptibility to mouse hepatitis virus A59 infection

  • Kim, Han-Woong;Seo, Sun-Min;Kim, Jun-Young;Lee, Jae Hoon;Lee, Han-Woong;Choi, Yang-Kyu
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.36.1-36.12
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    • 2021
  • Background: Mouse hepatitis virus (MHV) A59 is a highly infectious pathogen and starts in the respiratory tract and progresses to systemic infection in laboratory mice. The complement system is an important part of the host immune response to viral infection. It is not clear the role of the classical complement pathway in MHV infection. Objectives: The purpose of this study was to determine the importance of the classical pathway in coronavirus pathogenesis by comparing C1qa KO mice and wild-type mice. Methods: We generated a C1qa KO mouse using CRISPR/Cas9 technology and compared the susceptibility to MHV A59 infection between C1qa KO and wild-type mice. Histopathological and immunohistochemical changes, viral loads, and chemokine expressions in both mice were measured. Results: MHV A59-infected C1qa KO mice showed severe histopathological changes, such as hepatocellular necrosis and interstitial pneumonia, compared to MHV A59-infected wild-type mice. Virus copy numbers in the olfactory bulb, liver, and lungs of C1qa KO mice were significantly higher than those of wild-type mice. The increase in viral copy numbers in C1qa KO mice was consistent with the histopathologic changes in organs. These results indicate that C1qa deficiency enhances susceptibility to MHV A59 systemic infection in mice. In addition, this enhanced susceptibility effect is associated with dramatic elevations in spleen IFN-γ, MIP-1 α, and MCP-1 in C1qa KO mice. Conclusions: These data suggest that C1qa deficiency enhances susceptibility to MHV A59 systemic infection, and activation of the classical complement pathway may be important for protecting the host against MHV A59 infection.

Study on Antitumor Effects of Sambonggangyongbaneo-tang and Gagamsojeokbaekchoo-san (삼봉강룡반어탕(三蓬薑龍半魚湯)과 가감소적백출산(加減消積白朮散)의 항암효과(抗癌效果)에 대(對)한 비교연구(比較硏究))

  • Kim, Dong-Woo;Ko, Seung-Hi;Choi, You-Kyung;Shim, Mun-Ki;Yeo, Eun-Kyung;Park, Se-Ki;Park, Chong-Hyeong;Ko, Jae-Chul;Jun, Chan-Yong;Han, Yang-Hee;Lee, Chung-Jung-Hye
    • The Journal of Internal Korean Medicine
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    • v.21 no.3
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    • pp.425-432
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    • 2000
  • Objective : It is well known that Gagamsojeokbackchool-san show antitumor effects and its activities are result from enhancement of immune function, we investigated the antitumor effects of Sambonggangyongbaneo-tang and its mechanism. Methods : We measured change of body weight, weight of immune organs (Spleen, Thymus), Liver, Kidney, tumor weight, cytotoxicity for investigation of antitumor effects of Sambonggangyongbaneo-tang. Results : 1. The body weight of mouse has no significant difference between control and sample groups. 2. The weights of immune organs (Spleen and Thymus) decreased significantly in sample groups. The weights of Liver and Kidney have no significant difference. 3. The tumor weights in mouse decreased significantly in sample groups and showed dose-dependent effect. 4. Cell viability of Sarcoma 180 has no significant difference in sample groups. 5. HeLa cell viability has no significant difference in low concentration, but it decreased significantly in high concentration. Conclusions : According to the above results, it could be suggested that Sambonggangyongbaneo-tang has prominant antitumor effects and cytotoxicity.

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Effect of Junsibaekchulsan on the humoral and cell-mediated immune responses in mouse (전씨백출산(錢氏白朮散)이 생쥐의 체액성(體液性) 면역반응(免疫反應)과 세포성(細胞性) 면역반응(免疫反應)에 미치는 효과(效果))

  • Sim Mun-Kyeoung;Park Eun-Jeang
    • The Journal of Pediatrics of Korean Medicine
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    • v.8 no.1
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    • pp.39-58
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    • 1994
  • Even though appropriate immune response is necessary for the survival of the individual, excessive or insufficient immune response might cause autoimmune or allergic disease respectively. So the immune response must be controlled to the degree that is beneficial for the well being of the individual. This study was undertaken to know the effects of Junsibaekchulsan(JB) on the immune system od the mouse. For the evalulation of the cell-mediated immunity(CMI), delayed-type hypersensitivity against dinitrofluorobenzene(DNFB) were measured, and humoral immunity, hemagglutinin and hemolysin titers against SRBCs(sheep red blood cells) were measured, and rosette formation of spleen cells with SRBCs were measured. For the evaluation of innate immunity, phagocytic activity of macrophages, natural killer cell activity, and reactive nitrogen and oxygen intermediates were measured. The results are as follows: 1. The administration of JB depressed the antibody formation (hemagglutinin and hemolysin) against SRBCs. 2. The administration of JB did not affect the delayed-type hypersensitivity against DNFB. 3. The administration of JB did not affect the cytotoxic activity of natural killer cells. 4. The administration of JB increased the phagocytic activity of macrophages. 5. The administration of JB increased the rosette formating cells of the spleen cells. 6. The exposure of JB induced the secretion of reactive nitrogen intermediates but administration of JB deperssed the production of reactive oxygen intermediates. Administration of JB selectively depressed the humoral immune response without affecting CMI and innate immunity. These results of JB on the immune system might be useful for the treatment of such.

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Production of Monoclonal Antibody to Polychlorinated Biphenyl Induced Cytochrome P-450 LMII in Rat Liver (Polychlorinated Biphenyl에 의한 백서간 Cytochrome P-$450_{LMII}$에 대한 Monoclonal Antibody 생성에 관한 연구)

  • Kim, Jung-Hye;Kim, Jae-Ryong;Lee, Ki-Yung
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.103-110
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    • 1986
  • Cytochrome P-450(CP-450) is one of the three components of the liver microsomal enzyme system which hydroxylates fatty acids, hydrocarbons and a variety of drugs and other foreign compounds. Female Balb/c mice were immunized with purified polychlorinated bipheny(PCB)-induced CP-450 LMII. The spleen cells derived from immunized mice were fused with $SP^2$ myeloma cells using polyethylene glycol(PEG 3500). The hybrid cells were selected by hypoxanthine-aminopterine and thymidine(HAT) medium and the culture fluid were screened by enzyme-linked immunosorbent assay to CP450 LMII. The hybrid cess(${\times}10^7$) were innoculated into intraperitoneal cavity of Balb/c mice for the purpose of production of ascitic fluids. Monoclonal antibody(Mab) was purified from ascitic fluid by DEAE cellulose ion exchange chromatography and $I^{125}$-labeled Mab was also confirmed by autoradiography and SDS-polyacrylamide gel electrophoresis (MW : 55,000 and 110,000).

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The Effects of Antioxidant Vitamins Via Comet and Mitogenic Assay in Mice Exposed to $AFB_1$ ($AFB_1$에 노출된 마우스에서 Comet Assay와 Mitogenic Assay에 의한 항산화 비타민의 효과)

  • Park, Seon-Ja
    • Journal of Korean Biological Nursing Science
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    • v.3 no.2
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    • pp.21-33
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    • 2001
  • The objective of this study was to examine the effects of antioxidant vitamins on the cellular oxidant damage by observing the mitogenicity in the mouse spleen and the strand breaks of DNA in mouse blood induced by $AFB_2$. Intraperitoneal(i.p.) injections of vitamin C(VC) of 10 mg/kg and vitamin E(VE) of 63.8 mg/kg were repeatedly administered to male ICR mice of 6 weeks old at intervals of 4 times every 2 days. After one hour vitamin treatments, $AFB_1$ of 0.4 mg/kg was injected into the $AFB_2$ plus vitamin treated groups in the same way. On the other hands, into the $AFB_2$ only treated group, only $AFB_2$ was injected without vitamins in the same method as above. The results of the experiment are as follows ; as regard to comet assay, DNA strand breaks were clearly present and they formatted a typical comet tail in the mice blood of the $AFB_2$ only treated groups. However, comet tails apparently disappeared in $AFB_2$ plus antioxidant vitamins treated groups since oxidant damage was controlled in an almost similar level to the control group. Mitogenicity of the spleen also showed a similar tendency as before, and these differences were more remarkably observed in the reaction against Con-A, which is a T-cell mitogen. In these data, the statistical significance was p<0.01. The LDL and VLDL levels were 408.72, 504.47 mg/dl respectively in the $AFB_2$ only treated groups. Compared with the $AFB_1$ only treated groups, those of $AFB_2$ plus antioxidant vitamin treated groups decreased to 272.06(VC), 305.28 mg/dl(VE), respectively. On the other hand, HDL levels were diminished to 32.60, 29.60 mg/dl in $AFB_2$ only treated groups, compared to 42.23, 41.14 mg/dl in the $AFB_2$ plus antioxidant vitamins treated groups. But, blood glucose levels were not statistically significant.

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