Characterization of monoclonal antibodies against waterfowl parvoviruses VP3 protein

Virology Journal

Yun Zhang¹  Ming Liu¹  Yuhuan Chen¹  Xiaoying Wu¹  Hongyu Liu¹  Shuyi Tan²  Jinzhe Li¹  Youlan Gao¹  Shumei Zhang¹  Xiuchen Yin¹ 

[1] State Key Lab of State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin, 150001, China;Hainan Key Lab of Tropical Animal Reproduction & Breeding and Epidemic Disease Research, Haikou, 571100, China

关键词: VP3;    MDPV;    GPV;    Monoclonal Antibody;   

Others  :  1152953
DOI  :  10.1186/1743-422X-9-288

 received in 2012-03-23, accepted in 2012-10-30,  发布年份 2012

【 摘 要 】

Background

The VP3 protein of goose parvovirus (GPV) or Muscovy duck parvovirus (MDPV), a major structural protein, can induce neutralizing antibodies in geese and ducks, but monoclonal antibodies (MAbs) against VP3 protein has never been characterized.

Results

Three hybridoma cell lines secreting anti-GPV VP3 MAbs were obtained and designated 4A8, 4E2, and 2D5. Immunoglobulin subclass tests differentiated them as IgG2b (4A8 and 4E2) and IgG2a (2D5). Dot blotting assays showed that three MAbs reacted with His-VP3 protein in a conformation-independent manner. A competitive binding assay indicated that the MAbs delineated two epitopes, A and B of VP3. Immunofluorescence assay showed that MAbs 4A8, 4E2, and 2D5 could specifically bind to goose embryo fibroblast cells (GEF) or duck fibroblast cells (DEF) infected with GPV and MDPV. Dot blotting also showed that the MAbs recognized both nature GPV and MDPV antigen. Western blotting confirmed that the MAbs recognized VP3 proteins derived from purified GPV and MDPV particles. The MAbs 4A8 and 2D5 had universal reactivity to heterologous GPV and MDPV tested in an antigen-capture enzyme-linked immunosorbent assay.

Conclusions

Preparation and characterization of these the MAbs suggests that they may be useful for the development of a MAb-capture ELISA for rapid detection of both GPV and MDPV. Virus isolation and PCR are reliable for detecting GPV and MDPV infection, but these procedures are laborious, time-consuming, and requiring instruments. These diagnosis problems highlight the ongoing demand for rapid, reproducible, and automatic methods for the sensitive detection of both GPV and MDPV infection.

【 授权许可】

   
2012 Yin et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Gough RE: Goose parvovirus infection. Ames. Iowa: Iowa State University Press; 1991:684-690. [Dis Poultry]
• [2]Kisary J: Derzsy’s disease of geese. Vol 4 edition. Amsterdam: Elsevier; 1985:157-162. [Virus Infections of Vertebrates]
• [3]Takehara K, Nishio T, Hayashi Y, Kanda J, Sasaki M, Abe N, Hiraizumi M, Saito S, Yamada T, Haritani M: An outbreak of goose parvovirus infection in Japan. J Vet Med Sci 1995, 57:777-779.
• [4]Gough RE: Goose parvovirus infection. In Dis Poultry. 11th edition. Edited by Saif YM, Barnes HJ, Glisson JR, Fadly AM, McDougal LR, Swayne DE. Ames, IA: Iowa State Univ. Press; 2003:367-374.
• [5]Jansson DS, Feinstein R, Kardi V, Mató T, Palya V: Epidemiologic investigation of an outbreak of goose parvovirus infection in Sweden. Avian Dis 2007, 51:609-613.
• [6]Zádori Z, Erdei J, Nagy J, Kisary J: Characteristics of the genome of goose parvovirus. Avian Pathol 1994, 23:359-364.
• [7]Zádori Z, Stefancsik R, Rauch T, Kisary J: Analysis of the complete nucleotide sequences of goose and muscovy duck parvoviruses indicates common ancestral origin with adeno-associated virus 2. Virology 1995, 212:562-573.
• [8]Tatar-Kis T, Mato T, Markos B, Palya V: Phylogenetic analysis of Hungarian goose parvovirus isolates and vaccine strains. Avian Pathol 2004, 33:438-444.
• [9]Chu CY, Pan MJ, Cheng JT: Genetic variation of the nucleocapsid genes of waterfowl parvovirus. J Vet Med Sci 2001, 63:1165-1170.
• [10]Jestin V, Le Bras MO, Cherbonnel M, Le Gall G, Bennejean G: Isolement de virus de la maladie de Derzsy très pathogens chez le canard de Barbarie. Recueil de Mèd Vètèinaaire 1991, 167:849-857.
• [11]Le Gall-Reculé G, Jestin V: Biochemical and genomic characterization of Muscovy duck parvovirus. Arch Virol 1994, 139:121-131.
• [12]Le Gall-Reculé G, Jestin V, Chagnaud P, Blanchard P, Jestin A: Expression of muscovy duck parvovirus capsid proteins (VP2 and VP3) in a baculovirus expression system and demonstration of immunity induced by the recombinant proteins. J Gen Virol 1996, 77:2159-2163.
• [13]Poonia B, Dunn PA, Lu H, Jarosinski KW, Schat KA: Isolation and molecular characterization of a new Muscovy duck parvovirus from Muscovy ducks in the USA. Avian Pathol 2006, 35:435-441.
• [14]Gough RE: Application of the agar gel precipitation and virus neutralization tests to the serological study of goose parvovirus. Avian Pathol 1984, 13:501-509.
• [15]Wang CY, Shieh HK, Shieh JH KOCY, Chang PC: Expression of capsid proteins and non-structural proteins of waterfowl parvoviruses in Escherichia coli and their use in serological assays. Avian Pathol 2005, 34:376-382.
• [16]Kardi V, Szegletes E: Use of ELISA procedures for the detection of Derzsy’s disease virus of geese and of antibodies produced against it. Avian Pathol 1996, 25:25-34.
• [17]Zhang Y, Li YF, Liu M, Zhang DB, Guo DC, Liu CG, Zhi HD, Wang XM, Li G, Li N, Liu SG, Xiang WH, Tong GZ: Development and evaluation of a VP3-ELISA for the detection of goose and Muscovy duck parvovirus antibodies. J Virol Meth 2010, 163:405-409.
• [18]Takehara K, Hyakutake K, Imamura T, Mutoh K, Yoshimura M: Isolation, identification, and plaque titration of parvovirus from Muscovy ducks in Japan. Avian Dis 1994, 38:810-815.
• [19]Takehara K, Nakata T, Takizawa K, Limn CK, Mutoh K, Nakamura M: Expression of goose parvovirus VP1 capsid protein by a baculovirus expression system and establishment of fluorescent antibody test to diagnose goose parvovirus infection. Arch Virol 1999, 144:1639-1645.
• [20]Limn CK, Yamada T, Nakamura M, Takehara K: Detection of goose parvovirus genome by polymerase chain reaction: distribution of goose parvovirus in Muscovy ducklings. Virus Res 1996, 42:167-172.
• [21]Woźniakowski G, Samorek-Salamonowicz E, Kozdruń W: Quantitative analysis of waterfowl parvovirus in geese and Muscovy ducks by real-time polymerase chain reaction: correlation between age, clinical symptoms and DNA copy number of waterfowl prvoviruses. BMC Vet Re 2012, 15:8-29.
• [22]Alexandrov M, Alexandrova R, Alexandrov I, Zacharieva S, Lasarova S, Doumanova L, Peshev R, Donev T: Fluorescent and electron-microscopy immunoassays employing polyclonal and monoclonal antibodies for detection of goose parvovirus infection. J Virol Meth 1999, 79:21-32.
• [23]Ju HY, Wei N, Wang Q, Wang CY, Jing ZQ, Guo L, Liu DP, Gao MC, Ma B, Wang JW: Goose parvovirus structural proteins expressed by recombinant baculoviruses self-assemble into virus-like particles with strong immunogenicity in goose. Biochem and Bioph Res Commu 2011, 409:131-136.
• [24]Le Gall-Recule G, Jestin V: Biochemical and genomic characterization of muscovy duck parvovirus. Arch Virol 1994, 139:121-131.
• [25]Kaufmann B, Simpson AA, Rossmann MG: The structure of human parvovirus B19. Proc Natl Acad Sci USA 2004, 101:11628-11633. 28
• [26]Chipman PR, Agbandje-McKenna M, Kajigaya S, Brown KE, Young NS, Baker TS, Rossmann MG: Cryo-electron microscopy studies of empty capsids of human parvovirus B19 complexed with its cellular receptor. Proc Natl Acad Sci USA 1996, 93:7502-7506.
• [27]Schettler CH: Isolation of a highly pathogenic virus from geese with hepatitis. Avian Dis 1970, 15:323-325.
• [28]Liu M, Chen XD, Wang Y, Zhang Y, Li YF, Wang YF, Shen N, Chen H: Characterization of monoclonal antibodies against Muscovy duck reovirus σB protein. Virology J 2010, 7:133. BioMed Central Full Text
• [29]Wilson MB, Nakane PK: Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. North Holland, New York: Elsevier; 1978:215-224. [In immunofluorescence and related staining techniques]
• [30]Hou HS, Su YP, Shieh HK, Lee LH: Monoclonal antibodies against different epitopes of nonstructural protein σNS of avian reovirus S1133. Virology 2001, 282:161-175.