• 대한한의학회지
• /
• 제31권4호
• /
• pp.63-78
• /
• 2010

Objectives: This study was carried out to know the effects of Gwan-Jul-Bang-7 (hereafter referred to GJB-7) on the inhibition of arthritis induced by collagen on the mouse. Methods: To assess the effects of GJB-7 on mouse with arthritis induced by collagen II, we conducted several experiments such as analysis of cytotoxicity, hepatotoxicity, arthritis index, total cell number of draining lymph nodes and paw joints, value of immunocyte in PBMC (peripheral blood mononuclear cell), DLN (draining lymph node) and paw joint, measurement of cytokine and anti-collagen II, observation of the histological changes of joint. Results: 1. Cytotoxicity against HFC (human fibroblast cells) was not observed in any concentration and hepatotoxicity was not observed in the GJB-7 treated group. 2. The incidence of arthritis significantly decreased. 3. Total cell number of draining lymph nodes significantly increased and total cell number of paw joints significantly decreased. 4. The percentage of $CD8^+$ cells in PBMC (peripheral blood mononuclear cell) significantly increased. The percentage of $CD3^+/CD69^+$, and $CD3^+/CD49b^+$ cells in PBMC significantly decreased. 5. The percentage of $CD19^+,\;CD3^+$, and $CD4^+/CD25^+$ cells in DLN (draining lymph nodes) significantly increased. The percentage of $B220^+/CD23^+$ cells in DLN significantly decreased. 6. The percentage of $CD3^+,\;CD4^+$, and $CD11b^+/Gr-1^+$ cells in paw joints significantly decreased. 7. The production of TNF-$\alpha$, IL-6, and MCP-1 significantly decreased. 8. Anti-collagen II in serum significantly decreased. 9. With the hematoxylin and eosin stain, inflammation of joint decreased. Under Masson's trichrome stain, the cartilage destruction and synovial cell proliferation and the expression of collagen fibers decreased. Conclusions: Comparison of the results for this study showed that GJB-7 had immunomodulatory effects. So we expect that GJB-7 could be used as an effective drug for not only rheumatoid arthritis but also another auto-immune diseases.

• 대한수의학회지
• /
• 제29권2호
• /
• pp.33-39
• /
• 1989

Pure separation of various leukocytes is required for the assessment of their roles in immunological and phisiological function. In this study, pure separation of monocytes from canine peripheral blood was attempted. At first, mononuclear cells (PBMC) were separated by ficoll-hypaque gradient method and then monocytes were recovered from PBMC suspensions in sucrose gradient Sol. (PBMC-Sucrose), autologous plasma (PBMC-Plasma) and autologous serum (PBMC-Serum) incubated at $37^{\circ}C$ for 2 hours. 1. In the separation of PBMC by ficoll-hypaque gradient method in canine blood, higher relative centrifugal force (RCF) was required, as high as more than 1,300xg RCF for 40 minutes, for clear formation of PBMC layer than that in human blood as usually used 400xg RCF for 40 minutes. 2. In monocytes-separation from three PBMC suspensions following PBMC separation, recovery-, purity- and viability-rate of monocytes showed better results in PBMC-Plasma and PBMC-Serum than in PBMC-Sucrose suspension, particulary showing better results from PBMC suspensions performed by centrifugation at 1,500xg RCF for 40 minutes.

• Tuberculosis and Respiratory Diseases
• /
• 제42권6호
• /
• pp.823-830
• /
• 1995

연구배경: 결핵감염시 세포매개성 면역반응이 관여하여 말초혈액내 조력 T 세포가 감소하는 것으로 알려저왔다. 그러나 말초혈액내 표지물질을 가진 세포의 수적 감소가 세포매개성 면연반응의 저하라고 할 수있지 여부가 앞으로 해결해야 될 문제인 것으로 지적되고 있다. 이에 저자들은 폐결핵 환자들의 말초혈액에서 활성화된, 또는 활성화 되고 있은 세포의 수적변화를 관찰하기 위하여 본 연구를 시행하였다. 방법: 객담 결핵균 양성인 폐결핵 환자 22명을 대상으로 말초혈액에서 IL-2R, VLA-1, TLiSAI의 활성화 표지에 대한 단세포군 항체를 면역조직 화학법으로 측정하였다. 결과: 1) 폐결핵 환자의 말포혈액에서 $T_1$(+) 세포 및 그 아형들의 단위 부피당 절대수는 $T_4$(+) 세포는 유의하게 감소하였고 $T_8$(+) 세포는 증가하였다(p<0.05). 2) 폐결핵 환자에서 전체 T임파구의 비율은 감소되어 있으며 $T_4$(+) 세포, $T_8$(+) 세포비율은 각각 유의하게 감소, 증가하였다. 또한 $T_4(+)/T_8(+)$ 비율도 유의한 감소가 있었다(p<0.05). 3) 폐결핵 환자에서 activated T cell의 단위 부피당 절대수는 대조군에 비해 모두 유의한 증가를 보였다(p<0.05). 4) 폐결핵 환자에서 activated T cell 비율은, IL-2R, VLA-1, TLiSAI, 각각 6.45+1.56%, 7.64+1.34%, 10.45+1.16%로 모두 대조군에 비해 모두 유의한 증가를 보이며 특히 TLiSAI 항체가 가장 많이 관찰 되었다. 결론: 폐결핵 감염시 말초혈액 임파구의 일부만이 활성화되어 세포매개성 면역반응에 참여하는 것으로 사료된다.

• Restorative Dentistry and Endodontics
• /
• 제25권2호
• /
• pp.212-218
• /
• 2000

The recruitment of leukocytes to a site of inflammation is dependent on a complex interplay of a number of cytokines. Monocyte chemoattractant protein-1 (MCP-1) is a potent chemoattractant for monocytes, whereas interleukin-8 (IL-8) has chemotactic activity for neutrophils, lymphocytes, and basophils. The purpose of this study was to determine the effects of several microbes found in infected root canal systems on the production of inflammatoy cytokines, interleukin 8 and monocyte chemoattractant protein-1 from human peripheral blood mononuclear cells (PBMC). Monocytes isolated from peripheral blood were stimulated by group A streptococci (GAS, ATCC 19615), Enterococcus faecalis (ATCC 29212), Streptococcus mutans (ATCC 10449), Streptococcus sanguis (clinical isolate), and Candida albicans (ATCC 90029) respectively. Each of these bacteria induced dose-dependent induction in IL-8 and MCP-1 determined by ELISA. IL-8 production by each bacteria was decreased in the range of the microbe-to-PBMC ratios of 0.1-1.0. Group A streptococci was the week inducer of MCP-1 production. These results suggest that different oral pathogens induce specific dose-dependent patterns of cytokine release. Such patterns may provide a means of control of the type of immune celles particularly with regard to inflammatory leukocyte recruitment.

• Restorative Dentistry and Endodontics
• /
• 제25권1호
• /
• pp.63-70
• /
• 2000

The properties of ideal retrograde filling materials include the ability to seal the root canal system in three dimensions and well tolerated by periradicular tissues. Biocompatibility testing has been done mainly with cytotoxicity tests using cell culture. Little attention has been paid to the potential adverse influence on the inflammatory and immune reaction in the periapical tissue. The purpose of this study was to investigate the effects of retrograde filling materials on human mononuclear cells in vitro. Freshly mixed and set specimens from six materials (Z100, Tetric Ceram, Fuji II, Fuji II LC, F2000, Compoglass Flow, and ZOE) were eluated with cell culture medium for 24 hours. Cytotoxic effects of these extracts were evaluated by determining cell viability and enzyme activity using MTT and lactate dehydrogenase (LD). The production of inflammatoy bone resorptive cytokine, TNF-${\alpha}$ was measured from human peripheral blood mononuclear cells (PBMC) exposed to the extracts by means of Endogen Human TNF-${\alpha}$ ELISA kit (Wobrun, MA, U.S.A.). Eluates and diluted (1 : 10) eluates with cell culture medium from freshly mixed Fuji IT had cytotoxic effects on mononuclear cells using MTT and LD. However, eluates from set Fuji II were not cytotoxic. Eluates form set ZOE exhibited cytotoxicity with LD test. TNF-${\alpha}$ levels were high in eluates from freshly mixed Fuji II and Z100. Diluted eluates from freshly mixed Z100 and F2000 stimulated the production of TNF-${\alpha}$. However, there were no significant difference in TNF-${\alpha}$ levels compared to controls. These results indicate that some materials could possibly stimulate bone resorption in the periapical tissue by means of the production of bone resorptive cytokine.

• 한국미생물·생명공학회지
• /
• 제42권3호
• /
• pp.312-315
• /
• 2014

인체 면역증강제로 활용 가능성을 조사하기 위해 R. aquatilis AY2000 균이 생산하는 당단백질인 AYS의 면역세포에 대한 독성과 면역증강 효과를 조사하였다. 그 결과 AYS는 적혈구에 대한 용혈작용과 hPBMC에 대한 증식 또는 사멸효과는 나타내지 않았으나, 배양 중인 hPBMC를 응집시키는 효과를 나타내었다. 또한 AYS는 in vitro에서 hPBMC에 작용하여 염증성 cytokine인 IL-6, IFN-${\gamma}$, TNF-${\alpha}$와 IL-5 생성을 유도하였고, 생쥐에서 AYS는 alum에 비해 항-BSA의 생산력을 더욱 증가시켰다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권23호
• /
• pp.10339-10343
• /
• 2015

Breast cancer is the most common cancer among females worldwide and a most prevalent malignancy in Iranian women. Chronic stress may make an important contribution to cancer, especially in the breast. Numerous studies showed roles of neurotransmitters in the occurrence and progression of cancers which are mediated by their various types of receptors. This study was conducted to evaluate alterations in the expression profile of dopamine receptor genes in peripheral blood mononuclear cells (PBMC) as stress factors in breast cancer patients and the human breast cancer cell line (MCF-7). Peripheral blood samples were obtained from 30 patients and 30 healthy individuals. Total mRNA was extracted from PBMC and MCF-7 cells and RT-PCR was performed to confirm the presence of five dopamine receptors (DRD1-DRD5). Expression changes of dopamine receptor genes were evaluated by real time PCR. We observed that DRD2-DRD4 in PBMCs of breast cancer patients were increased compared to healthy individuals. In addition, all dopamine receptor subtypes but DRD1 were expressed in MCF-7 cells. Therefore, alterations of these receptors as stress factors should be assessed for selecting appropriate drugs such as D2-like agonists for treatment of breast cancer after performing complimentary tests. Determining the expression profile of dopamine receptor genes thus seems promising.

• IMMUNE NETWORK
• /
• 제13권3호
• /
• pp.94-101
• /
• 2013

Amorphous silica particles, whose applications are increasing in many biomedical fields, are known to be less toxic than crystalline silica. In this study, the inflammatory effects of amorphous silica nanoparticles were investigated using 30-nm amorphous silica nanoparticles and human peripheral blood mononuclear cells (PBMCs) or purified monocytes. As a result, production of IL-$1{\beta}$ and IL-8 were increased. In addition, the mitochondrial reactive oxygen species (ROS) was detected, which may lead to mitochondrial membrane disruption. Most importantly, inflammasome formation was observed. Therefore, these results provide immunological information about amorphous silica nanoparticles and suggest that amorphous silica nanoparticles can evoke innate immune reactions in human monocytes through production of IL-$1{\beta}$ and IL-8.

• 약학회지
• /
• 제54권3호
• /
• pp.192-199
• /
• 2010

Colorectal cancer is one of the most common alimentary malignancies. In this study, the antitumor activity of activated human natural killer (NK) cells against human colorectal cancer was evaluated in vivo. Human NK cells are the key contributors of innate immune response and the effective functions of these cells are enhanced by cytokines. Human peripheral blood mononuclear cells (PBMC) were cultured with interleukin-2 (IL-2)-containing medium for 14 days and resulted in enriched NK cell population. The resulting populations of the cells comprised 7% $CD3^+CD4^+$ cells, 25% $CD3^+CD8^+$ cells, 13% $CD3^-CD8^+$ cells, 4% $CD3^+$CD16/$CD56^+$ cells, 39% $CD3^+$CD16/$CD56^-$ cells, and 52% $CD3^-$CD16/$CD56^+$ cells. Tumor necrosis factor alpha (TNF-$\alpha$), interferon gamma (IFN-$\gamma$), IL-2, IL-4, and IL-5 transcripts of the activated NK cells were confirmed by RT-PCR. In addition, activated NK cells at doses of 2.5, 5 and 10 million cells per mouse inhibited 10%, 34% and 47% of SW620-induced tumor growth in nude mouse xenograft assays, respectively. This study suggests that NK cell-based immunotherapy may be used as an adoptive immunotherapy for colorectal cancer patients.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권4호
• /
• pp.2637-2641
• /
• 2013

6,8-Dihydroxy-7-methoxy-1-methyl-azafluorenone (DMMA), a purified compound from Polyalthia cerasoides roots, is cytotoxic to various cancer cell lines. The aims of this study were to demonstrate the type of cancer cell death and the mechanism(s) involved. DMMA inhibited cell growth and induced apoptotic death in human leukemic cells (HL-60, U937, MOLT-4), human breast cancer MDA-MB231 cells and human hepatocellular carcinoma HepG2 cells in a dose dependent manner, with $IC_{50}$ values ranging between 20-55 ${\mu}M$. DMMA also decreased cell viability of human peripheral blood mononuclear cells. The morphology of cancer cells induced by the compound after staining with propidium iodide and examined under a fluorescence microscope was condensed nuclei and apoptotic bodies. Mitochondrial transmembrane potential (MTP) was decreased after 24h exposure in all five types of cancer cells. DMMA-induced caspase-3, -8, and -9 activity was strongly induced in human leukemic HL-60 and MOLT-4 cells, while in U937-, MDA-MB231- and HepG2-treated cells there was partial induction of caspase. In conclusion, DMMA-induced activation of caspase-8 and -9 resulted in execution of apoptotic cell death in human leukemic HL-60 and MOLT-4 cell lines via extrinsic and intrinsic pathways.