旋毛虫重组二肽基肽酶1在体外对大鼠外周血单个核细胞增殖、吞噬、凋亡等的影响

doi:10.11843/j.issn.0366-6964.2020.019

Abstract

Abstract: This research aimed to investigate the effect of Trichinella spiralis dipeptidyl-peptidase 1 (Dpp1) on rat peripheral blood mononuclear cell (PBMC) proliferation, migration, apoptosis, nitric oxide secretion and phagocytosis in vitro. A pair of primers were designed based on Dpp1 gene nucleotides in GenBank, Dpp1 gene was cloned by RT-PCR and the expression vector was constructed to obtain the recombinant protein (rDpp1). The reactogenicity of rDpp1 was analyzed by Western blot. PBMCs were separated from rats and co-incubated with rDpp1 in vitro,binding of rDpp1 with rat PBMCs was confirmed by immunofluorescence assay (IFA), and the effects of different concentration of rDpp1 (10, 20, 40 μg·mL^-1) on cell proliferation,migration,nitric oxide production,phagocytosis and apoptosis were determined. The results showed that rDpp1 at a concentration of 40 μg·mL^-1 significantly promoted the proliferation of PBMCs; rDpp1 at 10 μg·mL^-1 could promote cell migration, while 20 and 40 μg·mL^-1 concentrations showed a significant promotion; rDpp1 with 40 μg·mL^-1 concentration significantly promoted NO secretion; rDpp1 with 10 μg·mL^-1 promoted cell phagocytosis, and 20 and 40 μg·mL^-1 concentrations showed very significant promotion; each concentration of proteins could significantly promote the apoptosis of PBMCs. The functions of rat PBMCs were influenced by T. spiralis dipeptidyl-peptidase 1 via different pathways.

Key words: Trichinella spiralis, dipeptidyl-peptidase 1, peripheral blood mononuclear cells, phagocytosis, apoptosis

CLC Number:

LIAO Shuyi, CHU Wen, XU Lixin, SONG Xiaokai, LI Xiangrui, YAN Ruofeng. Effect of Trichinella spiralis Recombinant Dipeptidyl-peptidase 1 on Proliferation, Phagocytosis, and Apoptosis of Rat PBMCs in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1362-1370.

References 33

[1]	朱兴全, 龚广学, 薛富汉, 等. 旋毛虫病[M]. 郑州:河南科学技术出版社, 1993.ZHU X Q, GONG G X, XUE F H, et al. Trichinelliasis[M]. Zhengzhou:Henan Science and Technology Press, 1993. (in Chinese)
[2]	MURRELL K D, POZIO E. Worldwide occurrence and impact of human trichinellosis, 1986-2009[J]. Emerg Infect Dis, 2011, 17(12):2194-2202.
[3]	POZIO E. World distribution of Trichinella spp. infections in animals and humans[J]. Vet Parasitol, 2007, 149(1-2):3-21.
[4]	RAWLINGS N D, BARRETT A J. Evolutionary families of peptidases[J]. Biochem J, 1993, 290(1):205-218.
[5]	KURIBAYASHI M, YAMADA H, OHMORI T, et al. Endopeptidase activity of cathepsin C, dipeptidyl aminopeptidase I, from bovine spleen[J]. J Biochem, 1993, 113(4):441-449.
[6]	TURK D, JANJIĆ V, STERN I, et al. Structure of human dipeptidyl peptidase I (cathepsin C):exclusion domain added to an endopeptidase framework creates the machine for activation of granular serine proteases[J]. EMBO J, 2001, 20(23):6570-6582.
[7]	SAJID M, MCKERROW J H. Cysteine proteases of parasitic organisms[J]. Mol Biochem Parasitol, 2002, 120(1):1-21.
[8]	BROMME D, NALLASETH F S, TURK B. Production and activation of recombinant papain-like cysteine proteases[J]. Methods, 2004, 32(2):199-206.
[9]	韩月. 旋毛虫组织蛋白酶B的克隆表达及在幼虫侵入小鼠肠上皮细胞中的作用[D]. 郑州:郑州大学, 2019.HAN Y. Cloning and expression of cathepsin B of Trichinella spiralis and its role in larval invasion of mouse intestinal epithelial cells[D]. Zhengzhou:Zhengzhou University, 2019. (in Chinese)
[10]	REBERNIK M, LENARČIČ B, NOVINEC M. The catalytic domain of cathepsin C (dipeptidyl-peptidase I) alone is a fully functional endoprotease[J]. Prot Exp Purif, 2019, 157:21-27.
[11]	HAN Y L, ZHOU A H, LU G, et al. DNA vaccines encoding Toxoplasma gondii cathepsin C 1 induce protection against toxoplasmosis in mice[J]. Korean J Parasitol, 2017, 55(5):505-512.
[12]	张萌萌. 旋毛虫肌幼虫与新生幼虫、雌性成虫与雄性成虫比较蛋白质组学研究[D]. 南京:南京农业大学, 2015.ZHANG M M. Comparative proteomics between muscle larvae and newborn larvae, female and male adult of Trichinella spiralis[D]. Nanjing:Nanjing Agricultural University, 2015. (in Chinese)
[13]	WANG W, YUAN C, WANG S, et al. Transcriptional and proteomic analysis reveal recombinant galectins of Haemonchus contortus down-regulated functions of goat PBMC and modulation of several signaling cascades in vitro[J]. J Proteomics, 2014, 98:123-137.
[14]	LI Y, YUAN C, WANG L K, et al. Transmembrane protein 147(TMEM147):another partner protein of Haemonchus contortus galectin on the goat peripheral blood mononuclear cells (PBMC)[J]. Parasit Vectors, 2016, 9:355.
[15]	YUAN C, ZHANG H, WANG W, et al. Transmembrane protein 63A is a partner protein of Haemonchus contortus galectin in the regulation of goat peripheral blood mononuclear cells[J]. Parasit Vectors, 2015, 8(1):211.
[16]	GADAHI J A, EHSAN M, WANG S, et al. Recombinant protein of Haemonchus contortus 14-3-3 isoform 2(rHcftt-2) decreased the production of IL-4 and suppressed the proliferation of goat PBMCs in vitro[J]. Exp Parasitol, 2016, 171:57-66.
[17]	KIM M Y, KIM J H, CHO J Y. Cytochalasin B modulates macrophage-mediated inflammatory responses[J]. Biomol Ther, 2014, 22(4):295-300.
[18]	胡孟娟, 周丽娜, 牛延萍, 等. 捻转血矛线虫半胱氨酸蛋白酶对山羊PBMCs免疫功能的影响[J]. 畜牧兽医学报, 2018, 49(4):804-810.HU M J, ZHOU L N, NIU Y P, et al. Effects of Haemonchus contortus cysteine protease on immune functions of goat PBMCs[J]. Acta Veterinaria et Zootechnica Sinica, 2018, 49(4):804-810. (in Chinese)
[19]	FANG L, SUN L, YANG J, et al. Heat shock protein 70 from Trichinella spiralis induces protective immunity in BALB/c mice by activating dendritic cells[J]. Vaccine, 2014, 32(35):4412-4419.
[20]	崔晶, 王中全, 李雍龙. 旋毛虫病疫苗[J]. 国外医学寄生虫病分册, 2004, 31(5):216-222.CUI J, WANG Z Q, LI Y L. Trichinellosis vaccine[J]. Foreign Medical Sciences Parasitic Diseases, 2004, 31(5):216-222. (in Chinese)
[21]	BAI X, HU X X, LIU X L, et al. Current research of trichinellosis in China[J]. Front Microbiol, 2017, 8:1472.
[22]	SUN X K, LI Y, NAQVI M A U H, et al. Succinate Coenzyme A ligase beta-like protein from Trichinella spiralis suppresses the immune functions of rat PBMCs in vitro and inhibits the secretions of interleukin-17in vivo[J]. Vaccines, 2019, 7(4):167.
[23]	吴玲燕, 王玉俭, 温玉玲, 等. 捻转血矛线虫NADH:泛醌氧化还原酶结构域包含蛋白基因的克隆表达及功能分析[J]. 畜牧兽医学报, 2017, 48(4):722-730.WU L Y, WANG Y J, WEN Y L, et al. Cloning, expression and function analysis of NADH:ubiquinone oxidoreductase domain containing protein in Haemonchus contortus[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(4):722-730. (in Chinese)
[24]	MOLL H. Dendritic cells and host resistance to infection[J]. Cell Microbiol, 2010, 5(8):493-500.
[25]	JACOBS H J, ASHMAN K, MEEUSEN E. Humoral and cellular responses following local immunization with a surface antigen of the gastrointestinal parasite Haemonchus contortus[J]. Vet Immunol Immunopathol, 1995, 48(3-4):323-332.
[26]	GILL H S. Cell-mediated immunity in merino lambs with genetic resistance to Haemonchus contortus[J]. Int J Parasitol, 1994, 24(5):749-756.
[27]	OLEKHNOVITCH R, BOUSSO P. Induction, propagation, and activity of host nitric oxide:lessons from Leishmania infection[J]. Trends Parasitol, 2015, 31(12):653-664.
[28]	MUÑOZ-CARRILLO J L, MUÑOZ-ESCOBEDO J J, MALDONADO-TAPIA C H, et al. Resiniferatoxin lowers TNF-α, NO and PGE2 in the intestinal phase and the parasite burden in the muscular phase of Trichinella spiralis infection[J]. Parasite Immunol, 2017, 39(1):e12393.
[29]	WEN Y L, WANG Y J, WANG W J, et al. Recombinant Miro domain-containing protein of Haemonchus contortus (rMiro-1) activates goat peripheral blood mononuclear cells in vitro[J]. Vet Parasitol, 2017, 243:100-104.
[30]	DONSKOW-SCHMELTER K, DOLIGALSKA M. Apoptosis, a protective mechanism for pathogens and their hosts[J]. Wiad Parazytol, 2005, 51(4):271-280.
[31]	PIEKARSKA J, GORCZYKOWSKI M, SZCZYPKA M, et al. The influence of immunosuppression on apoptosis and necrosis during experimental trichinellosis in mice[J]. Cent Eur J Immunol, 2012, 3:204-208.
[32]	WANG X L, LIU M Y, SUN S M, et al. An anti-tumor protein produced by Trichinella spiralis induces apoptosis in human hepatoma H7402 cells[J]. Vet Parasitol, 2013, 194(2-4):186-188.
[33]	YU Y R, DENG M J, LU W W, et al. Endoplasmic reticulum stress-mediated apoptosis is activated in intestines of mice with Trichinella spiralis infection[J]. Exp Parasitol, 2014, 145:1-6.

Effect of Trichinella spiralis Recombinant Dipeptidyl-peptidase 1 on Proliferation, Phagocytosis, and Apoptosis of Rat PBMCs in vitro

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