• Title/Summary/Keyword: Immune System

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Production and Breeding of Transgenic Cloned Pigs Expressing Human CD73

  • Lee, Seung-Chan;Lee, Haesun;Oh, Keon Bong;Hwang, In-Sul;Yang, Hyeon;Park, Mi-Ryung;Ock, Sun-A;Woo, Jae-Seok;Im, Gi-Sun;Hwang, Seongsoo
    • Development and Reproduction
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    • v.21 no.2
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    • pp.157-165
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    • 2017
  • One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-human-primate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were $127{\pm}18.9$. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as ${\alpha}-1,3-galactosyltransferase$ knock-out /hCD46 for xenotransplantation.

Functional Characteristics of Kefir as a Fermented Dairy Product: A Review (발효낙농유제품인 Kefir 다양한 기능 및 특성: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Song, Kwang-Young;Kim, Dong-Hyeon;Kim, Hong-Seok;Yim, Jin-Hyeok;Choi, Dasom;Hwang, Dae-Geun;Kim, Young-Ji;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.99-108
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    • 2013
  • Kefir is a unique fermented dairy product resulting from combined lactic acid and alcoholic fermentation of lactose in milk. Kefir is produced by the microbial activity of kefir grains. Kefir has numerous health benefits such as the reduction of cholesterol levels, stimulation of the immune system, antimutagenic and anticarcinogenic properties, and improvements of lactose intolerance. Furthermore, kefir is excellent as both a dietetic beverage and for protection against various diseases in small babies. Therefore, kefir has recently been regarded as an important functional dairy food. To date, the majority of research on kefir has focused on the applications of functional kefir using advanced biotechnology methods. The purpose of this review article is to facilitate the recognition of kefir as a novel functional food.

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The Radiation Protection effect of Tabebuia Avellanedae Extract on the Prostate in Male Rats (수컷 쥐 전립선에 대한 타히보 추출물의 방사선 방호효과 연구)

  • Jeon, Chan-hee;Kim, Jang-Oh;Lee, Yoon-Ji;Lee, Ji-Eun;Lee, Chang-Ho;Min, Byung-In
    • Journal of the Korean Society of Radiology
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    • v.14 no.6
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    • pp.755-762
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    • 2020
  • This study is desinged to examine the effects of Taheebo(Tabebuia avellanedae) extract on the prostate of male rats as a natural radiation protection agent. Taheebo extract is well known to inhibit cell growth for the cell lines of breast and prostate cancer. In this study, the X-ray 7 Gy was irradiated in the prostate of male rat to identify radiation protection effects by Taheebo Extracts, 1, 7, and 21 Days later, hematological changes, external toxicity assessments(LDH), antioxidant enzyme(SOD) activity changes and tissue change were observed. IR+TH group showed greater lymphocyte levels than the irradiation group, which is believed to affect the hematopoietic immune system's resilience. As a results of the external toxicity assessment, Taheebo extract's toxicity is maximum 18.128±5.16%, minimum 13.6945±4.43%. Taheebo is considered to be of little toxicity. The composition of prostate cell nuclei and cytoplasm in Control and TH group was honogeneous, whereas the cell nucleus cohesion in the prostate in irradiation group and inflammatory reactions in cytoplasm were shown. IR+TH group showed less inflammatory reactions of cytoplasm in the prostate than in the radiation irradiation group, but showed a cohesive phenomenon of cell nuclei. It is judged that Taheebo extract has radiation protection against prostate cells.

Glatiramer acetate inhibits the activation of NFκB in the CNS of experimental autoimmune encephalomyelitis (Glatiramer acetate 투여에 의한 자가면역성 뇌척수염 마우스의 중추신경계에서의 NFκB 활성 억제)

  • Hwang, Insun;Ha, Danbee;Kim, Dae Seung;Joo, Haejin;Jee, Youngheun
    • Korean Journal of Veterinary Research
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    • v.51 no.3
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    • pp.217-225
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    • 2011
  • Glatiramer acetate (GA; Copaxone) has been shown to be effective in preventing and suppressing experimental autoimmune encephalomyelitis (EAE), the animal model of multiple sclerosis (MS). It has been recently shown that GA-reactive T cells migrate through the blood-brain barrier, accumulate in the central nervous system (CNS), secrete antiinflammatory cytokines and suppress production of proinflammatory cytokines of EAE and MS. Development of EAE requires coordinated expression of a number of genes involved in the activation and effector functions of inflammatory cells. Activation of inflammatory cells is regulated at the transcriptional level by several families of transcription factors. One of these is the nuclear factor kappa B ($NF{\kappa}B$) family which is present in a variety of cell types and involved in the activation of immune-relative genes during inflammatory process. Since it is highly activated at site of inflammation, $NF{\kappa}B$ activation is also implicated in the pathogenesis of EAE. In this study, we examined whether the inhibition of $NF{\kappa}B$ activation induced by GA can have suppressive therapeutic effects in EAE mice. We observed the expression of $NF{\kappa}B$ and phospho-$I{\kappa}B$ proteins increased in GA-treated EAE mice compared to EAE control groups. The immunoreactivity in inflammatory cells and glial cells of $NF{\kappa}B$ and phospho-$I{\kappa}B$ significantly decreased at the GA-treated EAE mice. These results suggest that treatment of GA in EAE inhibits the activation of $NF{\kappa}B$ and phophorylation of $I{\kappa}B$ in the CNS. Subsequently, the inhibition of $NF{\kappa}B$ activation and $I{\kappa}B$ phosphorylation leads to the anti-inflammatory effects thereby to reduce the progression and severity of EAE.

Immunomodulating Activity of Crude Polysaccharide from Inonotus obliquus Sclerotia by Fractionation including MeOH Reflux

  • Lee, Kyung-Haeng;Kim, Hoon;Oh, Sung-Hoon;Hwang, Jong-Hyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.30 no.1
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    • pp.96-104
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    • 2017
  • To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysac- charide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer's patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production ($10{\mu}g/mL$) in Peyer's patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.

Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen

  • Patel, Maulikkumar;Gandotra, Vinod K.;Cheema, Ranjna S.;Bansal, Amrit K.;Kumar, Ajeet
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.9
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    • pp.1247-1255
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    • 2016
  • Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.

In silico Design of Discontinuous Peptides Representative of B and T-cell Epitopes from HER2-ECD as Potential Novel Cancer Peptide Vaccines

  • Manijeh, Mahdavi;Mehrnaz, Keyhanfar;Violaine, Moreau;Hassan, Mohabatkar;Abbas, Jafarian;Mohammad, Rabbani
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5973-5981
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    • 2013
  • At present, the most common cause of cancer-related death in women is breast cancer. In a large proportion of breast cancers, there is the overexpression of human epidermal growth factor receptor 2 (HER2). This receptor is a 185 KDa growth factor glycoprotein, also known as the first tumor-associated antigen for different types of breast cancers. Moreover, HER2 is an appropriate cell-surface specific antigen for passive immunotherapy, which relies on the repeated application of monoclonal antibodies that are transferred to the patient. However, vaccination is preferable because it would stimulate a patient's own immune system to actively respond to a disease. In the current study, several bioinformatics tools were used for designing synthetic peptide vaccines. PEPOP was used to predict peptides from HER2 ECD subdomain III in the form of discontinuous-continuous B-cell epitopes. Then, T-cell epitope prediction web servers MHCPred, SYFPEITHI, HLA peptide motif search, Propred, and SVMHC were used to identify class-I and II MHC peptides. In this way, PEPOP selected 12 discontinuous peptides from the 3D structure of the HER2 ECD subdomain III. Furthermore, T-cell epitope prediction analyses identified four peptides containing the segments 77 (384-391) and 99 (495-503) for both B and T-cell epitopes. This work is the only study to our knowledge focusing on design of in silico potential novel cancer peptide vaccines of the HER2 ECD subdomain III that contain epitopes for both B and T-cells. These findings based on bioinformatics analyses may be used in vaccine design and cancer therapy; saving time and minimizing the number of tests needed to select the best possible epitopes.

Association of Polymorphism Harbored by Tumor Necrosis Factor Alpha Gene and Sex of Calf with Lactation Performance in Cattle

  • Yudin, N.S.;Aitnazarov, R.B.;Voevoda, M.I.;Gerlinskaya, L.A.;Moshkin, M.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1379-1387
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    • 2013
  • In a majority of mammals, male infants have heavier body mass and grow faster than female infants. Accordingly, male offspring nursing requires a much greater maternal energy contribution to lactation. It is possible that the maternal-fetal immunoendocrine dialog plays an important role in female preparation for lactation during pregnancy. Immune system genes are an integral part of gene regulatory networks in lactation and tumor necrosis factor alpha ($TNF{\alpha}$) is a proinflammatory cytokine that also plays an important role in normal mammary gland development. The aim of this study was to evaluate the influence of the sex of calf and/or the -824A/G polymorphism in the promoter region of $TNF{\alpha}$ gene on milk performance traits in Black Pied cattle over the course of lactation. We also studied the allele frequency differences of -824A/G variants across several cattle breeds, which were bred in different climatic conditions. The G allele frequency decreased gradually over the course of lactation events in the Black Pied dairy cattle because of a higher culling rate of cows with the G/G genotype (p<0.001). In contrast to the genotypes A/A and A/G, cows with G/G genotype showed significant variability of milk and milk fat yield subject to sex of delivered calf. Milk yield and milk fat yield were significantly higher in the case of birth of a bull calf than with a heifer calf (p<0.03). The G allele frequency varies from 48% to 58% in Grey Ukrainian and Black Pied cattle to 77% in aboriginal Yakut cattle. Our results suggest that the $TNF{\alpha}$-824A/G gene polymorphism may have an influence on the reproductive efforts of cows over the course of lactation events depending on the sex of progeny. Allocation of resources according to sex of the calf allows optimizing the energy cost of lactation. This may be a probable reason for high G allele frequency in Yakut cattle breeding in extreme environmental conditions. Similarly, the dramatic fall in milk production after birth of a heifer calf increases the probability of culling for the cows with the G/G genotype in animal husbandry.

Effects of alfalfa flavonoids on the production performance, immune system, and ruminal fermentation of dairy cows

  • Zhan, Jinshun;Liu, Mingmei;Su, Xiaoshuang;Zhan, Kang;Zhang, Chungang;Zhao, Guoqi
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.10
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    • pp.1416-1424
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    • 2017
  • Objective: The objective of this study was to examine the effects of alfalfa flavonoids on the production performance, immunity, and ruminal fermentation of dairy cows. Methods: The experiments employed four primiparous Holstein cows fitted with ruminal cannulas, and used a $4{\times}4$ Latin square design. Cattle were fed total mixed ration supplemented with 0 (control group, Con), 20, 60, or 100 mg of alfalfa flavonoids extract (AFE) per kg of dairy cow body weight (BW). Results: The feed intake of the group receiving 60 mg/kg BW of AFE were significantly higher (p<0.05) than that of the group receiving 100 mg/kg BW. Milk yields and the fat, protein and lactose of milk were unaffected by AFE, while the total solids content of milk reduced (p = 0.05) linearly as AFE supplementation was increased. The somatic cell count of milk in group receiving 60 mg/kg BW of AFE was significantly lower (p<0.05) than that of the control group. Apparent total-tract digestibility of neutral detergent fiber and crude protein showed a tendency to increase (0.05<$p{\leq}0.10$) with ingestion of AFE. Methane dicarboxylic aldehyde concentration decreased (p = 0.03) linearly, whereas superoxide dismutase activity showed a tendency to increase (p = 0.10) quadratically, with increasing levels of AFE supplementation. The lymphocyte count and the proportion of lymphocytes decreased (p = 0.03) linearly, whereas the proportion of neutrophil granulocytes increased (p = 0.01) linearly with increasing levels of dietary AFE supplementation. The valeric acid/total volatile fatty acid (TVFA) ratio was increased (p = 0.01) linearly with increasing of the level of AFE supplementation, the other ruminal fermentation parameters were not affected by AFE supplementation. Relative levels of the rumen microbe Ruminococcus flavefaciens tended to decrease (p = 0.09) quadratically, whereas those of Butyrivibrio fibrisolvens showed a tendency to increase (p = 0.07) quadratically in response to AFE supplementation. Conclusion: The results of this study demonstrate that AFE supplementation can alter composition of milk, and may also have an increase tendency of nutrient digestion by regulating populations of microbes in the rumen, improve antioxidant properties by increasing antioxidant enzyme activities, and affect immunity by altering the proportions of lymphocyte and neutrophil granulocytes in dairy cows. The addition of 60 mg/kg BW of AFE to the diet of dairy cows was shown to be beneficial in this study.

Effect of Supplementing the Diet of Olive Flounder Paralichthys olivaceus with Sea Mustard Undaria pinnatifida Glycoprotein on Growth and the Immune System (사료 내 미역(Undaria pinnatifida) 당단백질의 첨가가 넙치(Paralichthys olivaceus) 치어의 성장 및 면역 증강에 미치는 영향)

  • An, Cheul-Min;Kim, Kang-Woong;Kim, Kyoung-Duck;Kim, Young-Min;Kim, In-Hye;Park, Su-Jin;Choi, Youn Hee;Nam, Taek Jeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.5
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    • pp.423-429
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    • 2012
  • This study evaluated the effects of adding sea mustard Undaria pinnatifida glycoprotein to the diet of juvenile olive flounder Paralichthys olivaceus on its growth, and levels of insulin-like growth factor I (IGF-I), IGF binding proteins (IGFBPs), and interleukins. Three experimental diets (U0, U0.5, and U1.0) were formulated that contained different amounts of an extract of U. pinnatifida (0, 0.5, and 1.0%, respectively). Experimental groups were established in triplicate (30 fish/group) and fed for 12 weeks. The experimental group fed 1.0% added U. pinnatifida glycoprotein had the greatest rate of weight gain, which differed significantly from the other experimental groups. SDS-PAGE of the plasma IGF-I and muscle protein showed that the experimental groups taking U. pinnatifida glycoprotein had significantly more IGF-I and a ca. 200 kDa protein, as compared to the control group. In addition, the amount of IGFBP-3 at ca. 43 kDa increased in the group given the U. pinnatifida extract, as compared to the control group. The interluekin-2, -4, -6, and -12 levels paralleled the level of growth factor in the groups given the U. pinnatifida extract. In conclusion, supplementing the diet of olive flounder with U. pinnatifida glycroprotein improved its growth and immunity.