Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (18): 3909-3917.doi: 10.3864/j.issn.0578-1752.2011.18.023

• RESEARCH NOTES • Previous Articles     Next Articles

Antigenic Expression and Distribution of Chitosan/pcDNA-DPV-gC Gene in the Vaccinated Ducklings

SHEN  Fu-Xiao, JIANG  Jin-Feng, CHENG  An-Chun, WANG  Ming-Shu, LU  Li-Ting, JIA  Ren-Yong, ZHU  De-Kang, CHEN  Xiao-Yue, SUN  Tao   

  1. 1.四川农业大学预防兽医研究所
    2.四川农业大学动物医学院禽病防治研究中心
    3.四川农业大学动物疫病与人类健康四川省重点实验室
    4.山东省出入境检验检疫局技术中心
  • Received:2011-11-30 Online:2011-09-15 Published:2011-05-10

Abstract: 【Objective】 DNA-chitosan nanoparticles carrying duck plague virus (DPV) gC gene were constructed by using complex coacervation process to investigate its antigenic expression and distribution in the vaccinated ducklings. 【Method】 The 20-day-old DPV-free ducklings were respectively immunized with chitosan/pcDNA-DPV-gC gene vaccine via intramuscular injection, nasal administration and oral administration. At intervals of 4 h, 12 h, 1 d, 3 d, 5 d, 7 d, 2 w, 4 w, 6 w and 10 w post-vaccination (p.v.), two ducklings were randomly euthanatized and their organs (liver, spleen, lung, kidney, pancreas, brain, thymus, Harderian gland, bursa of Fabricius, esophagus, duodenum, caecum, and rectum) were collected. Meanwhile, an indirect immunohistochemical staining (IHC) was developed to detect antigenic expression and distribution of DPV gC antigens in the vaccinated ducklings. 【Result】 The DPV gC proteins were found in the liver, bursa of Fabricius, duodenum, caecum and rectum in the intramuscular injection group at 1 d post-vaccination (p.v.); Moreover, the immunogenicity were firstly found in the lung at 12 h p.v., and the DPV gC proteins were observed in the bursa of Fabricius and Harderian gland in the nasal administration group at 1 d p.v. Furthermore, the positive signals were firstly found in the esophagus at 12 h p.v., and the DPV gC proteins were observed in the bursa of Fabricius, duodenum, caecum and rectum in the oral administration group at 1 d p.v.. The DPV gC proteins were distributed in the liver, lung, bursa of Fabricius, Harderian gland, esophagus, duodenum, caecum and rectum, which were served as the principal sites for DPV gC antigens localization. The positive immunogenicity was mainly distributed in the parenchymal hepatic cells, epithelial cells of lung, lymphocytes of Harderian gland and bursa of Fabricius, epithelial cells of esophagus, epithelial cells and lamina propria mucosae cells of intestinal tract. According to the immunogenicity intensity and duration time via different immunization routes, the positive staining in all tissues was in the order of intramuscular injection group>nasal administration group>oral administration group. 【Conclusion】 The results demonstrated that chitosan as a polycationic gene carrier could promote the expression efficiency of DPV gC antigens in the vaccinated ducklings. Intramuscular injection was considered to be the best immunization routes to inoculated chitosan/pcDNA- DPV-gC gene vaccine.

Key words:

[1]Converse K A, Kidd G A. Duck plague epizootics in the United States, 1967-1995. Journal of Wildlife Diseases, 2001, 37 (2): 347-357.

[2]Guo Y F, Cheng A C, Wang M S, Zhou Y. Purification of anatid herpesvirus 1 particles by tangential-flow ultrafiltration and sucrose gradient ultracentrifugation. Journal of Virological Methods, 2009, 161 (1): 1-6.

[3]Lian B, Xu C, Cheng A C, Wang M S, Zhu D K, Luo Q H, Jia R Y, Bi F J, Chen Z L, Zhou Y, Yang Z X, Chen X Y. Identification and characterization of duck plague virus glycoprotein C gene and gene product. Virology Journal, 2011, 7:349.

[4]Zou Q, Sun K F, Cheng A C, Wang M S, Xu C, Zhu D K, Jia R Y, Luo Q H, Zhou Y, Chen Z L, Chen X Y. Detection of anatid herpesvirus 1 gC gene by TaqManTM fluorescent quantitative real-time PCR with specific primers and probe. Virology Journal, 2010, 7:37.

[5]Herold B C, WuDunn D, Soltys N, Spear P G. Glycoprotein C of herpes simplex virus type 1 plays a principal role in the adsorption of virus to cells and in infectivity. Journal of Virology, 1991, 65 (3):1090-1098.

[6]Azab W, Tsujimura K, Maeda K, Kobayashi K, Mohamed Y M, Kato K, Matsumura T, Akashi H. Glycoprotein C of equine herpesvirus 4 plays a role in viral binding to cell surface heparan sulfate. Virus Research, 2010, 151(1):1-9.

[7]Osorio Y, Cohen J, Ghiasi H. Improved protection from primary ocular HSV-1 infection and establishment of latency using multigenic DNA vaccines. Investigative Ophthalmology & Visual Science, 2004, 45 (2): 506-514.

[8]Xiao S B, Chen H C, Fang L R, Liu C S, Zhang H, Jiang Y B, Hong W Z. Comparison of immune responses and protective efficacy of suicidal DNA vaccine and conventional DNA vaccine encoding glycoprotein C of pseudorabies virus in mice. Vaccine, 2004, 22 (3-4): 345-351.

[9]Liu F Y, Ma B, Zhao Y, Zhang Y, Wu Y H, Liu X M, Wang J W. Characterization of the gene encoding glycoprotein C of duck enteritis virus. Virus Genes, 2008, 37 (3): 328-332.

[10]Sato T, Ishii T, Okahata Y. In vitro gene delivery mediated by chitosan. effect of pH, serum, and molecular mass of chitosan on the transfection efficiency. Biomaterials, 2001, 22 (15): 2075-2080.

[11]Lee J I, Ha K S, Yoo H S. Quantum-dot-assisted fluorescence resonance energy transfer approach for intracellular trafficking of chitosan/DNA complex. Acta Biomaterialia, 2008, 4 (4):791-798.

[12]Shen F X, Ma G P, Cheng A C, Wang M S, Li C F, Sun K F, Chang H, Zhu D K, Jia R Y, Chen X Y, Sun T. Development and application of an indirect immunohistochemical method for the detection of duck plague virus vaccine antigens in paraffin sections and localization in the vaccinated duckling tissues. Poultry Science, 2010, 89 (9): 1915-1923.

[13]黎  敏, 朱德康, 卢  菲, 韩新峰, 程安春, 汪铭书, 刘晓东, 车 茜, 陈孝跃. 不同剂量小鹅瘟病毒VP3基因疫苗在雏鹅体内的表达时相和分布规律. 中国兽医科学, 2008, 38 (6): 514-520.

Li M, Zhu D K, Lu F, Han X F, Cheng A C, Wang M S, Liu X D, Che Q, Chen X Y. Distribution and expression kinetics of different doses of GPV-VP3 DNA vaccine in gosling. Chinese Veterinary Science, 2008, 38 (6): 514-520. (in Chinese)

[14]徐  超, 程安春, 汪铭书, 文  明, 刘  菲, 韩晓英, 宋  涌, 廖永洪, 陈孝跃. 间接酶免疫组化检测DPV在感染鸭体内细胞定位的研究和应用. 中国兽医学报, 2007, 27 (5): 640-644.

Xu C, Cheng A C, Wang M S, Wen M, Liu F, Han X Y, Song Y, Liao Y H, Chen X Y. Study and application on cellular localization of DPV antigens in infected ducks by indirect immunoperoxidase technique. Chinese Journal of Veterinary Science, 2007, 27 (5): 640-644. (in Chinese)

[15]张文龙, 殷  喆, 刘霓虹, 杨  涛, 刘胜旺, 步志高, 王君伟, 吴东来. 鸡白细胞介素2(IL-2)DNA-壳聚糖纳米粒的制备及体外转染. 农业生物技术学报, 2009, 17 (4): 547-553.

Zhang W L, Yin Z, Liu N H, Liu T, Liu S W, Bu Z G, Wang J W, Wu D L. Preparation of chicken interleukin-2 (IL-2) DNA-chitosan nanoparticles and its transfection in vitro. Journal of Agricultural Biotechnology, 2009, 17 (4): 547-553. (in Chinese)

[16]Ishii T, Okahata Y, Sato T. Mechanism of cell transfection with plasmid/chitosan complexes. Biochimica et Biophysica Acta, 2001, 1514 (1): 51-64.

[17]Liu W G, Sun S J, Cao Z Q, Zhang X, Yao K D, Lu W W, Luk K K D. An investigation on the physicochemical properties of chitosan/  DNA polyelectrolyte complexes. Biomaterials, 2005, 26(15): 2705-2711.

[18]Zheng F, Shi X W, Yang G F, Gong L L, Yuan H Y, Cui Y J, Wang Y, Du Y M, Li Y. Chitosan nanoparticle as gene therapy vector via gastrointestinal mucosa administration: results of an in vitro and in vivo study. Life Sciences, 2007, 80 (4): 388-396.

[19]Zhang X L, Yu C L, Shi X, Zhang C, Tang T T, Dai K R. Direct chitosan-mediated gene delivery to the rabbit knee joints in vitro and in vivo. Biochemical and Biophysical Research Communications, 2006, 341 (1): 202-208.

[20]Koping-Hoggard M, Tubulekas I, Guan H, Edwards K, Nilsson M, Varum K M, Artursson P. Chitosan as a nonviral gene delivery system. Structure-property relationships and characteristics compared with polyethylenimine in vitro and after lung administration in vivo. Gene Therapy, 2001, 8 (14): 1108-1121.

[21]沈福晓, 程安春, 汪铭书, 蒋金凤, 贾仁勇, 朱德康, 陈孝跃,    孙  涛, 杨金龙. 不同剂量鸭瘟病毒gC基因疫苗在雏鸭体内的抗原表达时相和分布规律. 畜牧兽医学报, 2010, 41 (6): 726-734.

Shen F X, Cheng A C, Wang M S, Jiang J F, Jia R Y, Zhu D K, Chen X Y, Sun T and Chen J L. Antigenic distribution and expression regularity in the different doses of pcDNA-DPV-gC DNA vaccine in Tianfu ducklings. Acta Veterinaria et Zoontechnica Sinica, 2010, 41 (6):726-734. (in Chinese)

[22]Mansouri S, Lavigne P, Corsi K, Benderdour M, Beaumont E, Fernandes J C. Chitosan-DNA nanoparticles as non-viral vectors in gene therapy: strategies to improve transfection efficacy. European Journal of Pharmaceutics and Biopharmaceutics, 2004, 57 (1): 1-8.

[23]Chen J, Yang W L, Li G, Qian J, Xue J L, Fu S K, Lu D R. Transfection of mEpo gene to intestinal epithelium in vivo mediated by oral delivery of chitosan-DNA nanoparticles. World Journal of Gastroenterology, 2004, 10 (1): 112-116.

[24]Kumar M, Behera A K, Lockey R F, Zhang J, Bhullar G, De La Cruz C P, Chen L C, Leong K W, Huang S K, Mohapatra S S. Intranasal gene transfer by chitosan-DNA nanospheres protects BALB/c mice against acute respiratory syncytial virus infection. Human Gene Therapy, 2002, 13 (12): 1415-1425.

[25]Ozbas-Turan S, Aral C, Kabasakal L, Keyer-Uysal M, Akbuga J. Co-encapsulation of two plasmids in chitosan microspheres as a non-viral gene delivery vehicle. Journal of Pharmacy and Pharmaceutical Sciences, 2003, 6 (1): 27-32.
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