目的抗菌肽LL-37(LL-37)是人源性阳离子抗菌18 000多肽(h CAP18)的成熟形式,对哮喘的发病具有一定的调节作用,但具体机制尚不清楚。文中旨在研究LL-37在诱导人嗜酸性粒细胞释放炎性递质中的作用,并探讨其影响哮喘发病的可能机制。方法选...目的抗菌肽LL-37(LL-37)是人源性阳离子抗菌18 000多肽(h CAP18)的成熟形式,对哮喘的发病具有一定的调节作用,但具体机制尚不清楚。文中旨在研究LL-37在诱导人嗜酸性粒细胞释放炎性递质中的作用,并探讨其影响哮喘发病的可能机制。方法选取2015年1月至2016年1月攀枝花学院附属医院呼吸科就诊的16例轻度或中度过敏性哮喘患者,16名健康志愿者。从研究对象的外周血中提取原代嗜酸性粒细胞,将细胞分别分为哮喘组和对照组。依据干预嗜酸性粒细胞的方式将哮喘组和健康组细胞分别分为:哮喘a亚组、对照a亚组;哮喘血小板活化因子(PAF)亚组、对照PAF亚组;哮喘15μg/m L LL-37亚组、对照15μg/m L LL-37亚组;哮喘30μg/m L LL-37亚组、对照30μg/m L LL-37亚组;哮喘IL-5亚组、对照IL-5亚组;哮喘IL-5+15μg/m L LL-37亚组、对照IL-5+15μg/m L LL-37亚组;哮喘IL-5+30μg/m L LL-37亚组、对照IL-5+30μg/m L LL-37亚组。依据嗜酸性粒细胞中添加抑制剂种类及抗菌肽将哮喘组和健康组细胞分别分为:哮喘b亚组、对照b亚组;哮喘PTX亚组、对照PTX亚组;哮喘WRW4亚组、对照WRW4亚组;哮喘苏拉明亚组、对照苏拉明亚组;哮喘LL-37亚组、对照LL-37亚组;哮喘PTX+LL-37亚组、对照PTX+LL-37亚组;哮喘WRW4+LL-37亚组、对照WRW4+LL-37亚组;哮喘苏拉明+LL-37亚组、对照苏拉明+LL-37亚组。ELISA检测各组细胞上清的半胱氨酸白三烯(Cysteinyl leukotrienes,Cys-LTs)浓度;Western blot分析健康组细胞内加入PTX、WRW4后PLA2、磷酸化c PLA2、磷酸化ERK水平变化以及白三烯刺激后h CAP18水平。结果 LL-37诱导对照组嗜酸性粒细胞15、30 min后,对照30μg/m L LL-37亚组Cys-LTs表达量较对照15μg/m L LL-37亚组均显著升高[(54.02±7.15)pg/105vs(37.86±6.33)pg/105、(53.30±6.99)pg/105vs(36.27±6.46)pg/105,P<0.05]。对照组嗜酸粒细胞中Cys-LTs表达量的比较,对照IL-5+15μg/m L LL-37亚组[(59.97±6.83)pg/105]、对照IL展开更多
Antimicrobial peptides(AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms.Cathelicidins are an important family of AMPs that exclusively exist...Antimicrobial peptides(AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms.Cathelicidins are an important family of AMPs that exclusively exist in vertebrates. Many cathelicidins have been identified from mammals, birds, reptiles and fish. To date, however, cathelicidins from amphibians are poorly understood. In the present study, two novel cathelicidins(OL-CATH1 and 2) were identified and studied from the odorous frog Odorrana livida.Firstly, the cDNAs encoding the OL-CATHs(780 and735 bp in length, respectively) were successfully cloned from a lung cDNA library constructed for the frog. Multi-sequence alignment was carried out to analyze differences between the precursors of the OL-CATHs and other representative cathelicidins.Mature peptide sequences of OL-CATH1 and 2 were predicted(33 amino acid residues) and their secondary structures were determined(OL-CATH1 showed a random-coil conformation and OL-CATH2 demonstrated α-helical conformation). Furthermore,OL-CATH1 and 2 were chemically synthesized and their in vitro functions were determined. Antimicrobial and bacterial killing kinetic analyses indicated that OL-CATH2 demonstrated relatively moderate and rapid antimicrobial potency and exhibited strong anti-inflammatory activity. At very low concentrations(10 μg/mL), OL-CATH2 significantly inhibited the lipopolysaccharide(LPS)-induced transcription and production of pro-inflammatory cytokines TNF-α, IL-1βand IL-6 in mouse peritoneal macrophages. In contrast, OL-CATH1 did not exhibit any detectableantimicrobial or anti-inflammatory activities. Overall,identification of these OL-CATHs from O. livida enriches our understanding of the functions of cathelicidins in the amphibian immune system. The potent antimicrobial and anti-inflammatory activities of OL-CATH2 highlight its potential as a novel candidate in anti-infective drug development.展开更多
Cathelicidins are a major family of antimicrobial peptides present in vertebrate animals with potent microbicidal and immunomodulatory activities. Four cathelicidins, namely fowlicidins 1 to 3 and cathelicidin B1, hav...Cathelicidins are a major family of antimicrobial peptides present in vertebrate animals with potent microbicidal and immunomodulatory activities. Four cathelicidins, namely fowlicidins 1 to 3 and cathelicidin B1, have been identified in chickens. As a first step to understand their role in early innate host defense of chickens, we examined the tissue and developmental expression patterns of all four cathelicidins. Real-time PCR revealed an abundant expression of four cathelicidins throughout the gastrointestinal, respiratory, and urogenital tracts as well as in all primary and secondary immune organs of chickens. Fowlicidins 1 to 3 exhibited a similar tissue expression pattern with the highest expression in the bone marrow and lung, while cathelicidin B1 was synthesized most abundantly in the bursa of Fabricius. Additionally, a tissue-specific regulatory pattern was evident for all four cathelicidins during the first 28 days after hatching. The expression of fowlicidins 1 to 3 showed an age-dependent increase both in the cecal tonsil and lung, whereas all four cathelicidins were peaked in the bursa on day 4 after hatching, with a gradual decline by day 28. An abrupt augmentation in the expression of fowlicidins 1 to 3 was also observed in the cecum on day 28, while the highest expression of cathelicidin B1 was seen in both the lung and cecal tonsil on day 14. Collectively, the presence of cathelicidins in a broad range of tissues and their largely enhanced expression during development are suggestive of their potential important role in early host defense and disease resistance of chickens.展开更多
Natural antimicrobial peptides (AMPs) are small cationic molecules that display antimicrobial activity against a wide range of bacteria, fungi and viruses. AMPs are multifunctional molecules that have an essential act...Natural antimicrobial peptides (AMPs) are small cationic molecules that display antimicrobial activity against a wide range of bacteria, fungi and viruses. AMPs are multifunctional molecules that have an essential activity in infection and inflammation: they play an important role in the innate immune response, not only as antimicrobial agents, but also as immunomodulating molecules and as an important link between the innate and adaptive immune response. In this article, we will discuss the antimicrobial activity, together with the novel properties of some of these molecules as immune modulators on the innate and adaptive immune response.展开更多
文摘目的抗菌肽LL-37(LL-37)是人源性阳离子抗菌18 000多肽(h CAP18)的成熟形式,对哮喘的发病具有一定的调节作用,但具体机制尚不清楚。文中旨在研究LL-37在诱导人嗜酸性粒细胞释放炎性递质中的作用,并探讨其影响哮喘发病的可能机制。方法选取2015年1月至2016年1月攀枝花学院附属医院呼吸科就诊的16例轻度或中度过敏性哮喘患者,16名健康志愿者。从研究对象的外周血中提取原代嗜酸性粒细胞,将细胞分别分为哮喘组和对照组。依据干预嗜酸性粒细胞的方式将哮喘组和健康组细胞分别分为:哮喘a亚组、对照a亚组;哮喘血小板活化因子(PAF)亚组、对照PAF亚组;哮喘15μg/m L LL-37亚组、对照15μg/m L LL-37亚组;哮喘30μg/m L LL-37亚组、对照30μg/m L LL-37亚组;哮喘IL-5亚组、对照IL-5亚组;哮喘IL-5+15μg/m L LL-37亚组、对照IL-5+15μg/m L LL-37亚组;哮喘IL-5+30μg/m L LL-37亚组、对照IL-5+30μg/m L LL-37亚组。依据嗜酸性粒细胞中添加抑制剂种类及抗菌肽将哮喘组和健康组细胞分别分为:哮喘b亚组、对照b亚组;哮喘PTX亚组、对照PTX亚组;哮喘WRW4亚组、对照WRW4亚组;哮喘苏拉明亚组、对照苏拉明亚组;哮喘LL-37亚组、对照LL-37亚组;哮喘PTX+LL-37亚组、对照PTX+LL-37亚组;哮喘WRW4+LL-37亚组、对照WRW4+LL-37亚组;哮喘苏拉明+LL-37亚组、对照苏拉明+LL-37亚组。ELISA检测各组细胞上清的半胱氨酸白三烯(Cysteinyl leukotrienes,Cys-LTs)浓度;Western blot分析健康组细胞内加入PTX、WRW4后PLA2、磷酸化c PLA2、磷酸化ERK水平变化以及白三烯刺激后h CAP18水平。结果 LL-37诱导对照组嗜酸性粒细胞15、30 min后,对照30μg/m L LL-37亚组Cys-LTs表达量较对照15μg/m L LL-37亚组均显著升高[(54.02±7.15)pg/105vs(37.86±6.33)pg/105、(53.30±6.99)pg/105vs(36.27±6.46)pg/105,P<0.05]。对照组嗜酸粒细胞中Cys-LTs表达量的比较,对照IL-5+15μg/m L LL-37亚组[(59.97±6.83)pg/105]、对照IL
基金supported by grants from the Jiangsu Students' Innovation and Entrepreneurship Training Program(2017suda098)the National Natural Science Foundation of China(31772455)+2 种基金Natural Science Foundation of Jiangsu Province(BK20160336 and BK20171214)Natural Science Foundation of College in Jiangsu Province(16KJB350004)Suzhou Science and Technology Development Project(SYN201504 and SNG2017045)
文摘Antimicrobial peptides(AMPs) are a group of gene-encoded small peptides that play pivotal roles in the host immune system of multicellular organisms.Cathelicidins are an important family of AMPs that exclusively exist in vertebrates. Many cathelicidins have been identified from mammals, birds, reptiles and fish. To date, however, cathelicidins from amphibians are poorly understood. In the present study, two novel cathelicidins(OL-CATH1 and 2) were identified and studied from the odorous frog Odorrana livida.Firstly, the cDNAs encoding the OL-CATHs(780 and735 bp in length, respectively) were successfully cloned from a lung cDNA library constructed for the frog. Multi-sequence alignment was carried out to analyze differences between the precursors of the OL-CATHs and other representative cathelicidins.Mature peptide sequences of OL-CATH1 and 2 were predicted(33 amino acid residues) and their secondary structures were determined(OL-CATH1 showed a random-coil conformation and OL-CATH2 demonstrated α-helical conformation). Furthermore,OL-CATH1 and 2 were chemically synthesized and their in vitro functions were determined. Antimicrobial and bacterial killing kinetic analyses indicated that OL-CATH2 demonstrated relatively moderate and rapid antimicrobial potency and exhibited strong anti-inflammatory activity. At very low concentrations(10 μg/mL), OL-CATH2 significantly inhibited the lipopolysaccharide(LPS)-induced transcription and production of pro-inflammatory cytokines TNF-α, IL-1βand IL-6 in mouse peritoneal macrophages. In contrast, OL-CATH1 did not exhibit any detectableantimicrobial or anti-inflammatory activities. Overall,identification of these OL-CATHs from O. livida enriches our understanding of the functions of cathelicidins in the amphibian immune system. The potent antimicrobial and anti-inflammatory activities of OL-CATH2 highlight its potential as a novel candidate in anti-infective drug development.
基金supported by a United States Department of Agriculture grant2008-35204-04544Oklahoma Center for the Advancement of Science and Technology grants HR07-113, and AR07.2-087, and Oklahoma Agricultural Experiment Station project H-2811
文摘Cathelicidins are a major family of antimicrobial peptides present in vertebrate animals with potent microbicidal and immunomodulatory activities. Four cathelicidins, namely fowlicidins 1 to 3 and cathelicidin B1, have been identified in chickens. As a first step to understand their role in early innate host defense of chickens, we examined the tissue and developmental expression patterns of all four cathelicidins. Real-time PCR revealed an abundant expression of four cathelicidins throughout the gastrointestinal, respiratory, and urogenital tracts as well as in all primary and secondary immune organs of chickens. Fowlicidins 1 to 3 exhibited a similar tissue expression pattern with the highest expression in the bone marrow and lung, while cathelicidin B1 was synthesized most abundantly in the bursa of Fabricius. Additionally, a tissue-specific regulatory pattern was evident for all four cathelicidins during the first 28 days after hatching. The expression of fowlicidins 1 to 3 showed an age-dependent increase both in the cecal tonsil and lung, whereas all four cathelicidins were peaked in the bursa on day 4 after hatching, with a gradual decline by day 28. An abrupt augmentation in the expression of fowlicidins 1 to 3 was also observed in the cecum on day 28, while the highest expression of cathelicidin B1 was seen in both the lung and cecal tonsil on day 14. Collectively, the presence of cathelicidins in a broad range of tissues and their largely enhanced expression during development are suggestive of their potential important role in early host defense and disease resistance of chickens.
文摘Natural antimicrobial peptides (AMPs) are small cationic molecules that display antimicrobial activity against a wide range of bacteria, fungi and viruses. AMPs are multifunctional molecules that have an essential activity in infection and inflammation: they play an important role in the innate immune response, not only as antimicrobial agents, but also as immunomodulating molecules and as an important link between the innate and adaptive immune response. In this article, we will discuss the antimicrobial activity, together with the novel properties of some of these molecules as immune modulators on the innate and adaptive immune response.
