饲料营养 Feed science and technology

抗菌肽Sublancin增强小鼠获得性免疫的研究

展开
  • 中国农业大学动物科技学院, 北京 100193
张晓雅(1995-),女,河北邢台人,硕士研究生,从事猪营养研究。E-mail:zhangxiaoya201695@163.com

收稿日期: 2017-07-12

  网络出版日期: 2018-01-04

基金资助

国家重点研发计划课题"新型饲用抗生素替代品及其综合应用技术研究"(2016YFD0501308)

Study of Antimicrobial Peptide Sublancin Enhance Acquired Immunity of Mice

Expand
  • College of Animal Science and Technology, China Agricultural University, Beijing 100193, China

Received date: 2017-07-12

  Online published: 2018-01-04

摘要

本试验旨在研究抗菌肽Sublancin灌胃对卵清白蛋白(OVA)免疫小鼠获得性免疫反应的影响。试验选用6周龄雌性BALA/c小鼠60只,随机分为5个组,每组12只小鼠。空白对照组小鼠灌胃生理盐水,阳性对照组小鼠灌胃2.5 mg/kg体重的左旋咪唑溶液,抗菌肽Sublancin组小鼠灌胃0.5、1.0和2.0 mg/kg体重的抗菌肽Sublancin溶液。各组小鼠连续灌胃14 d。在灌胃结束24 h后用模式抗原OVA皮下注射对各组小鼠进行免疫,14 d后加强免疫1次,二次免疫7 d后采血和取脾脏,用间接酶联免疫吸附测定(ELISA)法对血清OVA特异性免疫球蛋白G (IgG)及其亚类IgG1、IgG2a和细胞因子含量进行检测。结果表明:1)与空白对照组相比,0.5、1.0和2.0 mg/kg抗菌肽Sublancin组和阳性对照组小鼠血清OVA特异性IgG含量均显著或极显著提高(P<0.05或P<0.01)。2)与空白对照组相比,1.0和2.0 mg/kg抗菌肽Sublancin组和阳性对照组小鼠血清OVA特异性IgG1含量显著或极显著提高(P<0.05或P<0.01),1.0 mg/kg抗菌肽Sublancin和阳性对照组小鼠血清OVA特异性IgG2a含量显著提高(P<0.05),1.0 mg/kg抗菌肽Sublancin组和阳性对照组的小鼠脾脏淋巴细胞的刺激指数显著提高(P<0.05)。3)与空白对照组相比,1.0 mg/kg抗菌肽Sublancin组和阳性对照组的小鼠脾脏淋巴细胞上清液中1型辅助性T细胞(Th1)细胞因子干扰素-γ(IFN-γ)和白细胞介素-2(IL-2)的含量均显著或极显著提高(P<0.05或P<0.01)。4)与空白对照组相比,0.5和1.0 mg/kg抗菌肽Sublancin组和阳性对照组的小鼠脾脏淋巴细胞上清液中2型辅助性T细胞(Th2)细胞因子白细胞介素-4(IL-4)和白细胞介素-10(IL-10)的含量均显著或极显提高(P<0.05或P<0.01)。5)0.5、1.0和2.0 mg/kg抗菌肽Sublancin组与阳性对照组之间的以上检测指标均没有显著差异(P>0.05)。本试验结果表明,抗菌肽Sublancin可以诱导OVA免疫小鼠产生Th1和Th2混合型免疫反应,增强其体液免疫和细胞免疫功能。

本文引用格式

张晓雅, 杨青, 王帅, 杨天任, 曾祥芳, 谯仕彦 . 抗菌肽Sublancin增强小鼠获得性免疫的研究[J]. 动物营养学报, 2018 , 30(1) : 236 -245 . DOI: 10.3969/j.issn.1006-267x.2018.01.029

Abstract

This experiment was designed to investigate the effects of intragastric administration of antimicrobial peptide Sublancin on acquired immunity of mice immunized with ovalbumin (OVA). Sixty BALB/c mice were randomly assigned into 5 groups with 12 mice per group. Mice in the blank control group received normal saline by intragastric administration, mice in the positive control group received 2.5 mg/kg body weight (BW) levamisole by intragastric administration, and mice in the antimicrobial peptide Sublancin groups received 0.5, 1.0 and 2.0 mg/kg BW antimicrobial peptide Sublancin by intragastric administration, respectively. All the mice were administrated intragastric daily for 14 days. Mice were immunized with model antigen OVA by subcutaneous infection after the intragastric administration 24 h, and enhanced immunization 1 time after 14 days, took blood and spleen after the second immunization 7 days, the contents of OVA-specific immunoglobulin (IgG) and its ubtype IgG1, IgG2a and cytokine in serum were detected by indirect enzyme linked immunosorbent assay (ELISA) method. The results showed as follows:1) compared with the blank control group, the serum OVA-specific IgG content of mice in 0.5, 1.0 and 2.0 mg/kg antimicrobial peptide Sublancin groups and positive control group was significantly increased (P<0.05 or P<0.01). 2) Compared with the blank control group, the serum OVA-specific IgG1 content of mice in 1.0 and 2.0 mg/kg antimicrobial peptide Sublancin groups and positive control group was significantly increased (P<0.05 or P<0.01), the serum OVA-specific IgG2a content of mice in 1.0 mg/kg antimicrobial peptide Sublancin group and positive control group was significantly increased (P<0.05), the spleen lymphocytes stimulation index of mice in 1.0 mg/kg antimicrobial peptide Sublancin group and positive control group was significantly increased (P<0.05). 3) Compared with the blank control group, the contents of T helper type 1 cell (Th1) interferon-γ(IFN-γ) and interleukin-2 (IL-2) in spleen lymphocyte supernatant liquid of mice in 1.0 mg/kg antimicrobial peptide Sublancin group and positive control group were significantly increased (P<0.05 or P<0.01). 4) Compared with the blank control group, the contents of T helper type 2 cell (Th2) interleukin-4 (IL-4) and interleukin-10 (IL-10) in spleen lymphocyte supernatant liquid of mice in 0.5 and 1.0 mg/kg antimicrobial peptide Sublancin groups and positive control group were significantly increased (P<0.05 or P<0.01). 5) There were no significant difference on the above testing indices among 0.5, 1.0 and 2.0 mg/kg antimicrobial peptide Sublancin groups and positive control group (P>0.05). This study indicated that antimicrobial peptide Sublancin can induce the mixed immune responses of Th1 and Th2 of OVA immunized mice, and enhance the humoral immunity and cellular immunity function.

参考文献

[1] ZASLOFF M.Antimicrobial peptides of multicellular organisms[J].Nature,2002,415(6870):389-395.  

[2] LAI Y P,GALLO R L.AMPed up immunity:how antimicrobial peptides have multiple roles in immune defense[J].Trends in Immunology,2009,30(3):131-141.  

[3] NGUYEN L T,HANEY E F,VOGEL H J.The expanding scope of antimicrobial peptide structures and their modes of action[J].Trends in Biotechnoloy,2011,29(9):464-472.  

[4] ZHANG L J,GALLO R L.Antimicrobial peptides[J].Current Biology,2016,26:R14-R19.

[5] BALTZER S A,BROWN M H.Antimicrobial peptides-promising alternatives to conventional antibiotics[J].Journal of Molecular Microbiology and Biotechnology,2011,20(4):228-235.  

[6] MANSOUR S C,PENA O M,HANCOCK R E W.Host defense peptides:front-line immunomodulators[J].Trends Immunology,2014,35(9):443-450.  

[7] FRITZ J H,BRUNNER S,BIRNSTIEL M L,et al.The artificial antimicrobial peptide KLKLLLLLKLK induces predominantly a TH2-type immune response to co-injected antigens[J].Vaccine,2004,22(25/26):3274-3284.

[8] ZHANG H H,YANG X M,XIE Q M,et al.The Potent adjuvant effects of chicken β-defensin-1 when genetically fused with infectious bursal disease virus VP2 gene[J].Veterinary Immunology and Immunopathology,2010,136(1/2):92-97.

[9] ACOSTA J,CARPIO Y,VALDES I,et al.Co-administration of tilapia alpha-helical antimicrobial peptides with subunit antigens boost immunogenicity in mice and tilapia (Oreochromis niloticus)[J].Vaccine,2014,32(2):223-229.  

[10] OMAN T J,BOETTCHER J M,WANG H,et al.Sublancin is not a lantibiotic but an S-linked glycopeptides[J].Nature Chemical Biology,2011,7(2):78-80.  

[11] STEPPER J,SHASTRI S,LOO T S,et al.Cysteine S-glycosylation,a new post-translational modification found in glycopeptide bacteriocins[J].FEBS Letters,2011,585(4):645-650.  

[12] JI S Y,LI W L,BALOCH A R,et al.Improved production of sublancin via introduction of three characteristic promoters into operon clusters responsible for this novel distinct glycopeptides biosynthesis[J].Microbial Cell Factries,2015,14:17.

[13] PAIK S H,CHAKICHERLA A,HANSEN J N.Identification and characterization of the structural and transporter genes for,and the chemical and biological properties of Sublancin 168,a novel lantibiotic produced by Bacillus subtilis 168[J].The Journal of Biological Chemistry,1998,273(36):23134-23142.  

[14] WANG Q W,ZENG X F,S WANG S,et al.The bacteriocin Sublancin attenuates intestinal injury in young mice infected with Staphylococcus aureus[J].Anatomical Record,2014,297(8):1454-1461.  

[15] WANG G S,LI X,WANG Z.APD3.The antimicrobial peptide database as a tool for research and education[J].Nucleic Acids Research,2015,44:D1087-D1093.

[16] KOUWEN T R H M,TRIP E N,DENHAM E L,et al.The large mechanosensitive channel mscl determines bacterial susceptibility to the bacteriocin Sublancin 168[J].Antimicrobial Agents and Chemotherapy,2009,53(11):4702-4711.  

[17] YANG D,BIRAGYN A,HOOVER D M,et al.Multiple roles of antimicrobial defensins,cathelicidins,and eosinophil-derived neurotoxin in host defense[J].Annual Review of Immunology,2004,22(1):181-215.  

[18] NICHOLLS E F,MADERA L,HANCOCK R E W.Immunomodulators as adjuvants for vaccines and antimicrobial therapy[J].Annals of the New York Academy of Sciences,2010,1213:46-61.

[19] BOMMINENI Y R,PHAM G H,SUNKARA L T,et al.Immune regulatory activities of fowlicidin-1,a cathelicidin host defense peptide[J].Molecular Immunology,2014,59(1):55-63.  

[20] 曹雪涛.医学免疫学[M].6版.北京:人民卫生出版社,2013:49-110.

[21] 龚非力.医学免疫学[M].3版.北京:科学出版社,2009:167-168.

[22] FINKELMAN F D,HOLMES J,KATONA I M,et al.Lymphokine control of in vivo immunoglobulin isotype selection[J].Annual Review of Immunology,1990,8(1):303-333.  

[23] MOHAN T,SHARMA C,BHAT A A,et al.Modulation of HIV peptide antigen specific cellular immune response by synthetic α-and β-defensin peptides[J].Vaccine,2013,31(13):1707-1716.  

[24] KANG Y M,JIN H L,ZHENG G X,et al.The adjuvant effect of levamisole on killed viral vaccines[J].Vaccine,2005,23(48/49):5543-5550.

[25] MALIK F,SINGH J,KHAJURIA A,et al.A standardized root extract of Withania somnifera and its major constituent with an olide-A elicit humoral and cell-mediated immune responses by up regulation of Th1-dominant polarization in BALB/c mice[J].Life Sciences,2007,80(16):1525-1538.  

[26] GAUTAM M,SAHA S,BANI S,et al.Immunomodulatory activity of Asparagus racemosus on systemic Th1/Th2 immunity:implications for immunoadjuvant potential[J].Journal of Ethnopharmacology,2009,121(2):241-247.  

[27] ZHAI L J,LI Y T,WANG W Y,et al.Effect of oral administration of ginseng stem-and-leaf saponins(GSLS) on the immune responses to Newcastle disease vaccine in chickens[J].Vaccine,2011,29(31):5007-5014.  

[28] HANCOCK R E W,DIAMOND G.The role of cationic antimicrobial peptides in innate host defences[J].Trends in Microbiology,2000,8(9):402-410.  

[29] LILLARD J W,BOYAKA P N,CHERTOV O,et al.Mechanisms for induction of acquired host immunity by neutrophil peptide defensins[J].Proceedings of the National Academy of Sciences of the United States of America,1999,96(2):651-656.  

[30] TANI K,MURPHY W J,CHERTOV O,et al.Defensins act as potent adjuvants that promote cellular and humoral immune responses in mice to a lymphoma idiotype and carrier antigens[J].International Immunolog,2000,12(5):691-700.  

[31] AGGER E M,ROSENKRANDS I,OLSEN A W,et al.Protective immunity to tuberculosis with Ag85B-ESAT-6 in a synthetic cationic adjuvant system IC31[J].Vaccine,2006,24(26):5452-5460.  

[32] DE GONZALO C V G,ZHU L Y,OMAN T J,et al.NMR structure of the S-linked glycopeptides sublancin 168[J].ACS Chemical,Biology,2014,9(3):796-801.  

[33] BIRAGYN A,RUFFINI P A,LEIFER C A,et al.Toll-like receptor 4-dependent activation of dendritic cells by β-defensin 2[J].Science,2002,298(5595):1025-1029.  

[34] VAN DER DOES A M,JOOSTEN S A,VROOMANS E,et al.The antimicrobial peptide hLF1-11 drives monocyte-dendritic cell differentiation toward dendritic cells that promote antifungal responses and enhance Th17 polarization[J].Journal of Innate Immunity,2012,4(3):284-292.  

[35] PENA O M,AFACAN N,PISTOLIC J,et al.Synthetic cationic peptide IDR-1018 modulates human macrophage differentiation[J].PLoS One,2013,8(1):e52449.
文章导航

/