• Korean Journal of Acupuncture
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• v.22 no.2
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• pp.107-125
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• 2005

Objectives : The aim of this study was to investigate the asthma-suppressive and immuno-regulatory effect of NR-HA(Notopterygii Rhizoma Herbal-acupuncture) at Pyesu(BL13) on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental sloops, except the Normal group and the NR-HA group, were sensitized and challenged with OVA. The mice in the NR-HA group and the OVA-NR-HA group were treated with NR-HA(1%) at Pyesu(BL13). The mice in the OVA-Saline group were injected with saline at Pyesu(BL13). The mice in the OVA-Needle-prick group were treated with a single prick with an injection needle at Pyesu(BL13). NR-HA, saline injection and needle prick were administered for 8 weeks, three times a week Results : in vitro 1. The populations of granulocytes, $CD3e^-/CCR3^+$cells, $CD69^+/CD3e^+$ cells, $CD4^+\;cells\;and\;CD23^+/B220^+$ cells in the OVA-induced asthmatic mouse lungs decreased significantly by NR-HAS(Notopterygii Rhizoma Herbal-acupuncture solution). 2. The lung weight and total cells in lung of the OVA-NR-HA group decreased significantly compared with those of the OVA-Control group. 3. Total leukocytes and eosinophils in BALF of the OVA-NR-HA group decreas ed significantly compared with those of the OVA-Control group. 4. The collagen accumulation in the lung sections of the OVA-NR-HA group decreased significantly compared with that of the OVA-control group. 5. The concentrations of IL-4, IL-5, IL-13, IgE in BALF and serum of the OVA-NR-HA group decreased significantly compared with those of the OVA- Control group. 6. The numbers of $Gr-1^+/CD11b^+,\;CCR3^+,\;CD3e^+, \;CD19^+,\;CD3e^+/CD69^+$ cells in the OVA-NR-HA group decreased significantly compared with those of the OVA-Control group. 7. The mRNA expressions of $TNF-{\alpha}$, IL-5, IL-4 and IL-13 in lung of the OVA-NR-HA group decreased significantly compared with those of the OVA- Control group. 8. The NR-HA group did not show my considerable difference from the Normal group. The OVA-saline group and the OVA-Needle prick group showed suppressive effects on OVA-induced asthma however they were not statistically significant. Conclusion : These results suggest that NR-HA at Pyesu(BL13) is considered to be effective in treating asthma and to be put to practical use in the future asthma clinic.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.2
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• pp.203-211
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• 2020

Objective: Staphylococcus aureus (S. aureus) is one of the major microorganisms responsible for subclinical mastitis in dairy cattle. The present study was designed with the aim to explore the DNA methylation patterns using the Fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) techniques in a S. aureus-infected mouse model. Methods: A total of 12 out-bred Institute of Cancer Research female mice ranging from 12 to 13 weeks-old were selected to construct a mastitis model. F-MSAP analysis was carried out to detect fluctuations of DNA methylation between control group and S. aureus mastitis group. Results: Visible changes were observed in white cell counts in milk, percentage of granulocytes, percentage of lymphocytes, CD⁴⁺/CD⁸⁺ ratio (CD⁴⁺/CD⁸⁺), and histopathology of mice pre- and post-challenge with S. aureus. These findings showed the suitability of the S. aureus-infected mouse model. A total of 369 fragments was amplified from udder tissue samples from the two groups (S. aureus-infected mastitis group and control group) using eight pairs of selective primers. Results indicated that the methylation level of mastitis mouse group was higher than that in the control group. In addition, NCK-associated protein 5 (Nckap5) and transposon MTD were identified to be differentially methylated through secondary polymerase chain reaction and sequencing in the mastitis group. These observations might play an important role in the development of S. aureus mastitis. Conclusion: Collectively, our study suggests that the methylation modification in Nckap5 and transposon MTD might be considered as epigenetic markers in resistance to S. aureus-infected mastitis and provided a new insight into S. aureus mastitis research in dairy industry and public health.

• Applied Microscopy
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• v.32 no.3
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• pp.231-246
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• 2002

This study was carried out to investigate the effect of squalene (SQ) on the mouse hepatotoxicity induced by cadmium. ICR male mouse weighting about 30 gm were injected $CdCl_2$ (5.0 mg/kg) and SQ (180 mg/kg) into intraperitoneal. At the 1, 2, 3, 4, 5, 6, 7 days, livers were treated with superoxide dismutase (SOD) activity and transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows: SOD activity in the liver, Group A was higher than in normal. Group B was lower than in Group A. In the histological observation, nucleus of Group A showed irregular shape. Inner cavity of mitochondria swellen and development of cristae weakened. Swelling of Lamellae of rough endoplasmic reticulum (RER) showed. Nucleus of group B showed normal shape. Typical lamellae of RER were observed. These results described above treatment of SQ decreased the hepatotoxicity of the $CdCl_2$ and SOD activity in the mouse liver, and then it suggests SQ may be effective for the recovery of hepatic cell.

• Biomedical Science Letters
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• v.21 no.2
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• pp.60-68
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• 2015

NecroX-7 is a novel small compound of the NecroX series based on the indole moiety, which has potent cytoprotective and antioxidant properties. We previously detected potential immune regulatory effects of NecroX-7 in immune related diseases like Graft-versus-Host Disease. However, the function and the underlying mechanisms of immunological effects of NecroX-7 in the immune system have not been well established. In this study, we investigated the immune response characterization of differentially expressed genes of NecroX-7 administration in $CD4^+$ T cells by microarray analysis. $CD4^+$ T cells stimulated with NecroX-7 ($40{\mu}M$) or vehicle for 72 hours resulted in the identification of 337 differentially expressed genes (1.5 fold, P<0.05) by expression profiling analysis. Twenty eight of the explored NecroX-7-regulated genes were related to immune system processes. These genes were validated by quantitative real-time PCR. The most significant genes were glutathione reductase, eukaryotic translation elongation factor 1, lymphotoxin-alpha, heat shock protein 9 and chloride intracellular channel protein 4. These findings demonstrate the strongly immune response of NecroX-7 in $CD4^+$ T cells, suggesting that cytoprotection and immune regulation may underlie the critical aspects of NecroX-7 exposure.

• Laboraroty Animal Research
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• v.34 no.4
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• pp.302-310
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• 2018

CD47 (integrin-associated protein), a multi-spanning transmembrane protein expressed in all cells including red blood cells (RBCs) and leukocytes, interacts with signal regulatory protein ${\alpha}$ ($SIRP{\alpha}$) on macrophages and thereby inhibits phagocytosis of RBCs. Recently, we generated a novel C57BL/6J CD47 knockout ($CD47^{-/-}$ hereafter) mouse line by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease, and here report their hematological phenotypes. On monitoring their birth and development, $CD47^{-/-}$ mice were born viable with a natural male-to-female sex ratio and normally developed from birth through puberty to adulthood without noticeable changes in growth, food/water intake compared to their age and sex-matched wild-type littermates up to 26 weeks. Hematological analysis revealed a mild but significant reduction of RBC counts and hemoglobin in 16 week-old male $CD47^{-/-}$ mice which were aggravated at the age of 26 weeks with increased reticulocyte counts and mean corpuscular volume (MCV), suggesting hemolytic anemia. Interestingly, anemia in female $CD47^{-/-}$ mice became evident at 26 weeks, but splenomegaly was identified in both genders of $CD47^{-/-}$ mice from the age of 16 weeks, consistent with development of hemolytic anemia. Additionally, helper and cytotoxic T cell populations were considerably reduced in the spleen, but not in thymus, of $CD47^{-/-}$ mice, suggesting a crucial role of CD47 in proliferation of T cells. Collectively, these findings indicate that our $CD47^{-/-}$ mice have progressive hemolytic anemia and splenic depletion of mature T cell populations and therefore may be useful as an in vivo model to study the function of CD47.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.302-309
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• 2003

Background: CTLA4 (CD152), which is expressed on the surface of T cells following activation, has a much higher affinity for B7 molecules comparing to CD28, and is a negative regulator of T cell activation. In contrast to stimulating and agonistic capabilities of monoclonal antibodies specific to CTLA-4, CTLA4Ig fusion protein appears to act as CD28 antagonist and inhibits in vitro and in vivo T cell priming in variety of immunological conditions. We've set out to confirm whether inhibition of the CD28-B7 costimulatory response using a soluble form of human CTLA4Ig fusion protein would lead to persistent inhibition of alloreactive T cell activation. Methods: We have used CHO-$dhfr^-$ cell-line to produce CTLA4Ig fusion protein. After serum free culture of transfected cell line we purified this recombinant molecule by using protein A column. To confirm characterization of fusion protein, we carried out a series of Western blot, SDS-PAGE and silver staining analyses. We have also investigated the efficacy of CTLA4Ig in vitro such as mixed lymphocyte reaction (MLR) & cytotoxic T lymphocyte (CTL) response and in vivo such as experimental autoimmune encephalomyelitis (EAE), graft versus host disease (GVHD) and skin-graft whether this fusion protein could inhibit alloreactive T cell activation and lead to immunosuppression of activated T cell. Results: In vitro assay, CTLA4Ig fusion protein inhibited immune response in T cell-specific manner: 1) Human CTLA4Ig inhibited allogeneic stimulation in murine MLR; 2) CTLA4Ig prevented the specific killing activity of CTL. In vivo assay, human CTLA4Ig revealed the capacities to induce alloantigen-specific hyporesponsiveness in mouse model: 1) GVHD was efficiently blocked by dose-dependent manner; 2) Clinical score of EAE was significantly decreased compared to nomal control; 3) The time of skin-graft rejection was not different between CTLA4Ig treated and control group. Conclusion: Human CTLA4Ig suppress the T cell-mediated immune response and efficiently inhibit the EAE, GVHD in mouse model. The mechanism of T cell suppression by human CTLA4Ig fusion protein may be originated from the suppression of activity of cytotoxic T cell. Human CTLA4Ig could not suppress the rejection in mouse skin-graft, this finding suggests that other mechanism except the suppression of cytotoxic T cell may exist on the suppression of graft rejection.

• The Journal of Korean Medicine
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• v.23 no.4
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• pp.169-185
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• 2002

Objectives: This study was carried out to investigate 1he effects of Sasammaickmoondong-tang(SME) on colonic mucosal lesions induced by indomethacin in mouse. Methods: The normal group is 1hat no inflammation elicitated mouse. Control group is that gastro-inflammation elicitated mouse. Sample group is that SME administered mouse after gastro-inflammation elicitation. Results: In the common morphology and histochemical change, control group was observed various injury-mucous surface cell, micro-villi, paneth cell, surface epithelial cell, goblet cell-by hemorrhagic erosion, while sample group was as same as normal group. In the immunohistochemical change, 1he distributions of COX-1, Bcl-2, and BrdU treated with SME noticeably increased than control group(P<0.05). The distributions of TUNEL, $NF-{\kappa}B$, COX-2, $IL-2R-\alpha$, NK-1.1, ICAM-1, and CD11b/18 treated with SME noticeably decreased than control group(P<0.05). And the distribution of SBA was as same as normal group. Conclusions: According to the above results, it is supposed that Sasammaickmoondong-tang is applicable to colonic mucosal lesions.

• Preventive Nutrition and Food Science
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• v.8 no.1
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• pp.7-12
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• 2003

This study examined the effect of pine needle distillate (Pinus densiflora Sieb. et Zucc) on the immune system and hematological parameters. C57BL/6 male mice weighing 20 ~21 g were divided into 3 groups and intraperitonially injected with either 200 $\mu$L of saline (control), 50% diluted (P50) or 100% pine needle distillate (P100) once a day for 24 days. At the end of the experiment, the mice were anesthetized by ether and peripheral blood was collected from the femoral artery and the spleen was excised. Spleen weight decreased significantly (p<0.001) in the pine needle groups compared to the control group. The blood was used for a complete blood count and flow cytometrical analysis after immunofluorescence staining. The pine needle distillate dose-dependently decreased the CD4$^{＋}$/CD8 sup ＋/ ratio (p <0.05), and showed a tendency to increase the mean FSC (forward scatter) values of the CD8$^{＋}$T cells, while decrease the values of the CD4$^{＋}$T cells. There were no significant differences in WBC, RBC and platelet counts among the three groups, but hemoglobin and hemoglobin-related parameters and platelet volume increased and red blood cell volumes decreased with the administration of the pine needle distillate. These results suggest that the pine needle distillate may have immunosuppressive effects.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.18 no.1
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• pp.27-49
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• 2005

당귀음자가감방(當歸飮子加減方)과 외치방(外治方) 병용의 아토피 치료 기전을 규명하고자 NC/Nga 생쥐의 동물 병태 모델을 이용하여 다양한 면역 반응을 관찰하였던 바, 다음과 같은 결론을 얻었다. 1. NC/Nga 생쥐의 피부손상 정도 16주와 20부에 대조군에 비해 36.0%, 37.8% 감소하다. 2. NC/Nga 생쥐의 혈중(血中) IgE, IL-4, IL-5, IL-6, IgM, IgG1 및 IgG2a 수준은 대조군에 비하여 유의성 있게 감소하였고, IL-13 수준은 대조군에 비하여 감소하였으나 유의성을 나타내지 않았다. 반면, $IFN-{\gamma}$ 수준은 유의성 있게 증가하다. 3. NC/Nga 생쥐의 비장 무게는 대조군에 비하여 유의성있게 감소하였다. 4. NC/Nga 생쥐의 lymph node에서 B/f ratio는 증가된 대조군에 비하여 감소하였으며, $CD4^+$와 $CD8^+$ 세포 발현은 대조군에 비하여 증가하였고, $CD4^+$는 유의성있는 감소를, $CD8^+$는 유의성 없는 약간의 증가를 나타내었다. $CD69^+$, CD11a 세포 발현은 대조군에 비하여 유의성있게 감소하였다. 5. NC/Nga 생쥐의 피부조직배양에서 IL-4 IL-5, CCR3 유전자 발현은 대조군에 비하여 현저히 감소하였고, IL-6, IL-13, $CD69^+/CD3{\varepsilon}^+,{\;}CD19^+/CD44^+$ 발현량은 유의성있게 감소하였으며, $IFN-{\gamma}$의 유전자 발현은 대조군에 비하여 증가하였다. 6. NC/Nga 생쥐 귀, 목의 피부 조직 변화에서는 표피와 진피의 염증 정도와 침윤된 염증 면역세포 등이 대조군에 비하여 현저하게 감소되었다. 7. Lymphokine assay에서 IL-4 발현량은 대조군에 비하여 유의성 있게 감소(減少)하였고, $IFN-{\gamma}$의 발현량은 유의성 있게 증가하였다.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.3815-3819
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• 2012

The CD4+CD25+ regulatory T cell (Treg) is a special kind of T cell subset. Studies have showed that Treg cells are involved in a number of physiological processes and pathologic conditions such as autoimmune diseases, transplantation tolerance and cancer. Tregs with unique capacity for immune inhibition can impair anti-tumour immunity and help tumor cells to escape from immune surveillance. The aim of our study was to investigate whether Tregs are involved in hepatocellular carcinoma (HCC). A BABL/C mouse with HCC in situ model was established to evaluate the Treg existence in carcinoma tissues and the changes of Tregs in spleen using flow cytometry and immunohistochemistry methods. Granzyme B expression in carcinoma tissues was analyzed by immunohistochemistry to investigate the tumor local immune status.The proportion of CD4+CD25+/CD4+ spleen lymphocytes of tumor bearing mice ($18.8%{\pm}1.26%$) was found to be significantly higher than that in normal mice ($9.99%{\pm}1.90%$) (P<0.01 ). Immunohistochemistry of spleen tissue also confirmed that there was an increase in Treg in tumor-bearing mice, while in carcinomas it showed Treg cells to be present in tumor infiltrating lymphocyte areas while Granzyme B was rarely observed. Anti-tumour immunity was suppressed, and this might be associated with the increase of Tregs. Our observations suggest that the CD4+CD25+Treg/CD4+ proportion in spleen lymphocytes can be a sensitive index to evaluate the change of Tregs in hepatocellular carcinoma mice and the Treg may be a promising therapeutic target for cancer.