• Title, Summary, Keyword: 비장세포

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시험관 내 면역 기술에 의한 항체 생산에 관한 연구

  • 전태훈;서동상
    • The Korean Journal of Zoology
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    • v.37 no.1
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    • pp.19-32
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    • 1994
  • 가장 효과적인 H-2b 항원에 대한 시험관내 항체 생산 조건을 찾기 위하여 근교계 생쥐인 C57BL/6BySnj의 비장세포를 UV로 불활성학 시킨 후 항원으로 사용하고, A/wySnJ$\times$Sm/J(ASmJF1, hybrid)의 비장세포를 항원 수용자 계통으로 하여 5-7일동안 배양기에서 항체 생산을 유도하였다. 본 실험에서 T 임파구 대식세포와 임파구 분화 촉진 인자인Concanavalin A Lipopolvsaccharide. Pokeweed mitogen 등을 사용하여 20가지 조건으로 실험을 수행하여, 항체 생성 여부는 보체 의존성 세포 장애 실험과 면역 효소법에 의해 조사하였다 그 결과 모든 조건에서 항체생산이 확인되었으며. 가장 좋은 시험관내 항체 생산 조건으로는 T 임파구와 대식세포를 함께 사용하여 면역시킨 것이 가장 효과적이었다. 이 방법을 이용하여 항체 생산을 유도한 후 5일째 면역된 비장세포를 Sp2/0-Ag 14와 세포 융합시켜 H-2b 마우스의 체포 표면 항원에 대한 단일군항체 생산을 시도하였다. 또한 생체내 면역 방법과 비교하기 위해 6주간 C57BL/6BySnJ의 비장세포를 복강내에 주사하여 같은 조건으로 세포융합을 시도하였다. 그 결과 H-2b 세포의 표면 항원에 대한 항체 생산을 하는 세포군은 시험관내 면역 방법에서 3개 생체내 면역 방법에서 4개부 확인되었다.

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Effects of Agaricus blazei Murill Water Extract on Immune Response in BALB/c Mice (신령버섯(Agaricus blazei Murill) 열수 추출물의 면역 활성에 미치는 영향)

  • Kang, In Soon;Kim, Rang Ie;Kim, Gwang Sub;Kim, Na Ri;Shin, Joong Yup;Kim, Chaekyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1629-1636
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    • 2015
  • The edible mushroom Agaricus blazei Murill is known to have many physiological functions, including antitumor, antiviral, and anti-inflammatory effects. Aqueous extracts were obtained by extracting A. blazei in water at $90^{\circ}C$ for 15 h, followed by spray-drying with dextran at a 70:30 ratio. In this study, we examined the immunomodulatory effect of A. blazei Murill water extract (ABM) in BALB/c mice. Mice were administered orally with 4, 20, and 100 mg/kg of ABM for 21 days. ABM-treated mice did not show significant differences in body and organ weights compare to saline-treated control mice. Splenocytes isolated from ABM-administered mice revealed similar levels of cellularity and proliferation compared to control mice, whereas they showed increased natural killer (NK) cell activity and decreased IL-4 and IL-12 production. Different from in vivo results, splenocytes isolated from normal mice showed increased proliferation and $INF-{\gamma}$ production following ABM treatment in vitro. In addition, ABM treatment enhanced macrophage proliferation and nitric oxide (NO) production in a dose-dependent manner. However, ABM had no effect on LPS-induced NO production. These results suggest that A. blazei modulates immune function by increasing NK cell activity and macrophage function.

Comparison Study of Immunomodulatory Activity of Polysaccharide and Ethanol Extracted from Sargassum fulvellum (참모자반 조다당 추출물과 에탄올 추출물의 대식세포 및 비장세포 활성 비교)

  • Byun, Eui-Hong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1621-1628
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    • 2015
  • The immune system plays an important role in maintaining and protecting human health. In the present study, comparison of immuno-modulatory activities between polysaccharides (SFP) and ethanol (SFE) extracts separated from Sargassum fulvellum in macrophages and murine splenocytes were investigated. Immuno-modulatory activities of macrophages were estimated based on cell proliferation, nitric oxide (NO), inducible NO synthase (iNOS), and cytokine production in RAW 264.7 macrophage cells, and lipopolysaccharide was used as a positive control. SFP and SFE treatment did not affect cytotoxicity in RAW 264.7 macrophage cells, and SFP treatment significantly increased NO and cytokine production ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$), whereas SFE did not contribute to the increase in NO and cytokine production. In the case of splenocytes, SFP treatment increased splenocyte proliferation and also highly increased production of Th-1 type cytokines (IL-2 and $IFN-{\gamma}$) than those of SFE. Through this study, we confirmed that immuno-modulatory activities of Sargassum fulvellum may be due to polysaccharide extracts and this can be a potential nutraceutical.

사료 중 크릴 분말이 살모넬라 LPS로 자극한 브로일러의 비장세포와 PBMC 증식에 미치는 영향

  • 임진택;박인경;김재환;고태송
    • Proceedings of the Korea Society of Poultry Science Conference
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    • pp.94-95
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    • 2002
  • In order to evaluate the effect of dietary krill meal on immunity of broilers, the proliferation of splenocyte and PBMC (pheripheral blood mononuclear cell) from broilers fed experimental diets containing 0.0, 0.5, 1.0 and 2.0 o/o krill meal, respectively, and injected the Salmonella typhymurium lipopolysaccharide (LPS) were assayed. The proliferation of splenocyte was increased with the dietary krill levels, but was decreased with the LPS immunlogical stress. Con A addition in the medium increased the proliferation of the splenocytes from birds fed dietary krill or stimulated by LPS. In 21 day old broilers, dietary krill meal and addition of Con A decreased the proliferation of PBMC while enhanced proliferation of PBMC was shown in birds stressed by the LPS during 2nd week of age. The results indicated dietary krill meal affected immune response in broiler.

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Mitigation effects of red Platycodon grandiflorum extract on lipopolysaccharide-induced inflammation in splenocytes isolated from mice (홍도라지 추출물이 마우스에서 분리한 비장세포에서 lipopolysaccharide로 유도된 염증에 미치는 영향)

  • Park, Eun-Jung;Lee, You-Suk;Jeong, Hyun Cheol;Lee, Sung-Hyen;Lee, Hae-Jeung
    • Journal of Nutrition and Health
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    • v.52 no.3
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    • pp.243-249
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    • 2019
  • Purpose: Platycodon grandiflorum (PG) is known to have effective antimicrobial and anticancer activity. The main bioactive components of PG are saponins, and these could contribute to anti-inflammatory activity. However, little is known about the anti-inflammatory effect of PG. In this study, we aim to assess the anti-inflammatory response to Red PG Extract (RPGE) in splenocytes under ex vivo conditions. Methods: The cell viability of isolated splenocytes taken from mice was analyzed by performing a Cell Counting Kit-8 assay. The productions of nitric oxide (NO) and cytokines (specifically interleukin-6 (IL-6) and interleukin-10 (IL-10)) were measured utilizing Griess reagent and ELISA, respectively. Results: We found that co-treatment with RPGE and Lipopolysaccharide (LPS) decreased isolated splenocyte proliferation as compared with that of the LPS-stimulated control. We also observed that RPGE markedly suppressed NO synthesis and IL-6 production that was induced by LPS. There were no significant differences of IL-10 production between co-treatment with RPGE plus LPS and treatment with LPS alone. Conclusion: When taken together, our data has shown that RPGE mitigates LPS-induced inflammation in splenocytes isolated from mice. Further research is surely needed to confirm the anti-inflammation effects of RPGE in an in vivo model.

Immunomodulatory Activities of Ethanol Extract of Cordyceps militaris in Immunocompromised Mice (밀리타리스 동충하초(Cordyceps militaris) 에탄올 추출물의 면역억제 마우스 면역활성에 미치는 영향)

  • Kim, Hye-Ju;Lee, Tae-Ho;Kwon, Yong-Sam;Son, Mi-Won;Kim, Chae-Kyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.4
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    • pp.494-500
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    • 2012
  • In order to determine the functional benefits of $Cordyceps$ $militaris$ in the immune system, we examined the immunomodulatory activities of $Cordyceps$ $militaris$ in an immunocompromised C57BL/6 mice model. Mice were injected intraperitoneally with an immunosuppressive drug, cyclophosphamide, and then administered orally with 3% hydroxypropylmethylcellulose or 30, 100, and 300 mg/kg of 50% ethanol extract of $Cordyceps$ $militaris$ (CM 30, CM 100, and CM 300, respectively) for 12 days. Mice treated with CM displayed significantly increased splenocyte proliferation and natural killer cell activity compared to immunosuppressed control mice (p<0.05). The spleen cells isolated from mice treated with CM also displayed increased production of Th1 cytokines, including IL-2, IL-12, IFN-${\gamma}$ and TNF-${\alpha}$, suggesting enhanced cellular immunity in response to CM. However, CM had no significant effect on the production of IL-4 and IL-10. These results indicate that $Cordyceps$ $militaris$ enhances immune function by promoting immune cell proliferation and Th1 cytokine production.

Ulmus Macrocarpa Water Extract Prolongs Splenocyte Life Span (Ulmus Macrocarpa 열수 추출물에 의한 비장세포 수명 연장)

  • Kang, Kyung-Hwa;Hyun, Sook Kyung;Hwang, Hye Jin;Kim, Byoung Woo;Kim, Cheol Min;Chung, Kyung Tae;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1176-1183
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    • 2015
  • Ulmus macrocarpa has been used in Korean medicinal food material to physical disorder or tonic material. The purpose of the present study was to evaluate splenocyte life span expansion effects of Ulmus macrocarpa water extract (UMWE) in general cell culture condition. Splenocytes were handled in the presence of 100 μg/ml UMWE for several different time conditions. Live cells were detected with Hoechst 33,342 dye and cell survival molecules were identified through Western blot. Changes in level of cytokine synthesis were evaluated by ELISA. UMWE showed an effect on increased splenocyte survival. UMWE elevated slightly PI3K phosphorylation and ERK1/2 phosphorylation used at 48 hr and 96 hr. Moreover, Bcl-2 was elevated at 48 hr and 96 hr in UMWE-treated splenocytes. UMWE decreased caspase-3 level at 48 hr and 96 hr. ICAD protein increased at 48 hr culturing time. Hematopoietin IL-2 cytokine was down-regulated, however IL-4 hematopoietin cytokine was up-regulated in UMWE treated cell culture media. Increased IFN-γ levels were verified in the supernatant of UMWE-treated cells in all periods (48 hr and 96 hr). Increased patterns in the production of IL-12 cytokine occurred as compared with control after 48 and 96 hr in UMWE-treated-cell cultures. These results suggested that UMWE can prolong splenocyte life span by controlling various signal factors and cytokines.

Passive Immunity by Splenocyte Transfer against Amebic Meningoeneephalitis in Mice (세포에 의한 아메바성 수막뇌염에 대한 피동면역의 전달)

  • 임경일;유재숙
    • The Korean Journal of Parasitology
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    • v.26 no.3
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    • pp.169-174
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    • 1988
  • The role of passive cell-mediated transfer of immunity against primary amoebic meningoen- cephalitis(PAME) in mice was studied. Waegleria fowleri, ITMAP 359, were cultured in CGVS medium. The ICR mice used were six week-old males of average weight of 15 g. Immunization was done by three intraperitoneal injections of $1{\times}10^6$ N. fowleri trophozoites at the interval of one week. Splenocytes were obtained from normal and immune mice spleens, and Ix107 cells were administered intraperitoneally into mice 3 days before challenge infection. Mice were infected intranasally with $7{\times}10^4$ N. fowleri trophozoites in a $3{\;}{\mu}l$ suspension under secobarbiturate anesthesia. Transplants of normal or immune splenocytes seem to alter the pattern of the PAME level- opment. The splenocytcs transferred from immune mice reduced the mortality rate in the JV. fowleri infected mice, as compared with the mice transferred with the same number of normal splenocytes or without splenocyte, The blastogenic response of the splenocytes to both lipopoly- saccharide and concanavalin A was elevated on duty 7 after infection the mice transinoculated with immune splenocytes. The serum antibody titers in the mice transferred with immune spleno- cytes were increased gradually from day 7 up to day 20 after infections by mean of ELISA. It is suggested that the transfer of splenocytes from immuniged mice conferred immunity against N. fowleri infection.

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Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts (괭생이 모자반 추출물의 비장세포 면역활성 증강 효과)

  • Kim, Dong-Sub;Sung, Nak-Yun;Han, In-Jun;Lee, Byung-Soo;Park, Sang-Yun;Nho, Eun Young;Eom, Ji;Kim, Geon;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.52 no.6
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    • pp.515-528
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    • 2019
  • Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

Roles of Spleen Cells in the Regulation of Progesterone and IGF -I Secretion in the Hanwoo Luteal Cells (한우 황체세포의 Progesterone 및 IGF-I 분비에 대한 비장세포의 역할)

  • 성환후;민관식;박진기;박성재;양병철;이장형;장원경
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.105-111
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    • 1999
  • The effects of exogenous spleen cells on the progesterone and insulin like-growth factor-I (IGF-I) secretions in luteal cells were studied by using in vitro luteal cell culture system in the Hanwoo luteal cells. The corpora lutea(CL) were collected and pooled from the Korean native cattle(Hanwoo) ovaries from a local slaughter house. After enzymatic dissociation, combined large and small luteal cells(LLC and SLC)(1.0$\times$10$^{6}$ cells/$m\ell$) were incubated in D-MEM media containing antibiotics and 10% FCS. Spleen cells (1.0$\times$10$^{6}$ cells/$m\ell$) obtained from castrated adult male Hanwoo were added to luteal cells and co-cultured for 24 h in the absence or presence of luteinizing hormone (LH) (100 ng). Progesterone contents from luteal tissues were increased at CL-3 stage during each stage of estrous cycle. Progesterone secretion from luteal cell culture by the presence of LH (100 ng/$m\ell$) was positively stimulated compared with control. However, progesterone secretion was not changed by the addition of 5, 10 and 20% of spleen cells in the absence of LH. Co-culture of luteal cells with 10% of spleen cells in the presence of LH(l00ng/$m\ell$) significantly. enhanced after 24 h of culture. IGF-Isecretion from in vitro luteal cells co-culture by the addition of spleen cells (5%, 10% and 20%) was not significantly effected. Besides, in the presence of LH (100ng/$m\ell$), IGF-Isecretions from luteal cells by addition of spleen cells were higher than control media. However, LH alone significantly increased IGF-I secretion at 24 h of culture. These data provide the demonstrate that spleen cells can enhance LH action so as to stimulate progesterone secretion from Hanwoo luteal cells but have no effect to stimulate IGF-I secretion.

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