Supachart PANNEUM, Wilairat CHUMSING, Theera RUKKWAMSUK

Cocultivation of caprine arthritis encephalitis virus-infected macrophages with primary goat synovial cells

Primary goat synovial cells were developed to be cocultivated with caprine arthritis encephalitis virus (CAEV)-infected monocytes. Two CAEV-seropositive goats and one CAEV-seronegative goat were confirmed for viral particles in the blood using the PCR method. Thereafter, heparinized blood samples from the three goats were collected and isolated for monocytes, which were further cultivated to differentiate into macrophages. Cultivated macrophages of 7-10 days old were cocultivated with the primary goat synovial cells for 7 days. Microscopic examination was performed every 2 days for multinucleated syncytial formation, and immunofluorescence assay was applied using monoclonal antibody against CAEV on day 7 to detect viral particles. Results showed that the primary goat synovial cells cultivated with CAEV-infected macrophages from the two seropositive goats had multinucleated syncytial formation with positive immunofluorescence, while the cocultured macrophages from the seronegative goats showed normal appearance with negative immunofluorescence. The multinucleated syncytial cells were prepared for transmission electron microscope examination and the results indicated that CAEV particles were clearly identified. In conclusion, our developed primary goat synovial cells and the cultivating system were proven to be an appropriate isolation assay for CAEV, which was the first report of cultivation and isolation of CAEV in goats in Thailand.

Keywords:

Caprine arthritis encephalitis virus cocultivation, goat, macrophage,

PDF

___

Adams DS, Crawford TB, Klevjer-Anderson P. A pathogenetic study of the early connective tissue lesions of viral caprine arthritis-encephalitis Am J Pathol 1980; 99: 257-278.
Peterhans E, Greenland T, Badiola J, Harkiss G, Bertoni G, Amorena B, Eliaszewicz M, Juste RA, Krassnig R, Lafonti JP et al. Routes of transmission and consequences of small ruminant lentiviruses (SRLVs) infection and eradication schemes. Vet Res 2004; 35: 257-274.
de Andres D, Klein D, Watt NJ, Berriatua E, Torsteinsdottir S, Blacklaws BA, Harkiss GD. Diagnostic tests for small ruminant lentiviruses. Vet Microbiol 2005; 107: 49-62.
Adams DS, Klevjer-Anderson P, Carlson JL, McGuire TC, Gorham JR. Transmission and control of caprine arthritis- encephalitis virus. Am J Vet Res 1983; 44: 1670-1675.
Jolly PE, Narayan O. Evidence for interference, coinfections, and intertypic virus enhancement of infection by ovine- caprine lentiviruses. J Virol 1989; 63: 4682-4688.
Konishi M, Tsuduka S, Haritani M, Murakami K, Tsuboi T, Kobayachi T, Yosikawa K, Kimura KM, Sentsui S. An epidemic of caprine encephalitis virus: isolation of the virus. J Vet Med Sci 2004; 66: 911-917.
Lin TN. A serological survey and study on the effect of caprine arthritis-encephalitis virus infection on reproductive performance of goats in the western part of Thailand. MSc, Chulalongkorn University, Bangkok, Thailand, 2011.
Ratanapob N, Rukkwamsuk T. Culture and identification of goat synovial membrane cells for caprine arthritis encephalitis virus diagnosis. J Anim Vet Adv 2012; 11: 1827-1830.
Narayan O, Clement JE, Strandberg JD, Cork LC, Griffin DE. Biological characterization of the virus causing leukoencephalitis and arthritis in goats. J Gen Virol 1980; 50: 69-79.
Clavijo A, Thorsen J. Application of polymerase chain reaction for the diagnosis of caprine arthritis-encephalitis. Small Rum Res 1996; 22: 69-77.
Milhau N, Renson P, Dreesen I ,Greenland T, Bellaton C, Guiguen F, Mornex JF, Le Jan C. Viral expression and leukocyte adhesion after in vitro infection of goat mammary gland cells with caprine arthritis-encephalitis virus. Vet Immunol Immunopathol 2005; 103: 93-99.
Chantakot S, Watthanakul M. A study of carprine arthritis encephalitis virus antibodies in goats at a farm in Ratchaburi province. Pamphlet of Cattle Region 2005; 7-9: 58-69.
Ratanapob N. Caprine arthritis encephalitisvirus infection in goats raised in the central and western part of Thailand. MSc, Kasetsart University, Bangkok, Thailand, 2010.
Narayan O, Griffin DE, Silverstein AM. Slow virus infection: Replication and mechanisms of persistence of visna virus in sheep. J Infect Dis 1977; 135: 800-806.
Chebloune Y, Sheffer D, Karr BM, Stephens E, Narayan O. Restrictive type of replication of ovine/caprine lentiviruses in ovine fibroblast cell cultures. Virology 1996; 222: 21-30.
Narayan O, Cork LC. Caprine arthritis-encephalitis virus. In: Dinter Z, Morein B, editors. Virus Infections of Ruminants. 1st ed. Amsterdam, the Netherlands: Elsevier; 1990. pp. 441-452.
L’Homme Y, Ouardani M, Lévesque V, Bertoni G, Simard C, Pisoni G. Molecular characterization and phylogenetic analysis of small ruminant lentiviruses isolated from Canadian sheep and goats. Virol J 2011; 8: 271.
Pisoni G, Bertoni G, Puricelli M, Maccalli M, Moroni1 P. Demonstration of coinfection with and recombination by caprine arthritis-encephalitis virus and maedi-visna virus in naturally infected goats. J Virol 2007; 8: 4948-4955.
Petursson G, Nathanson N, Georgsson G, Panitch H, Palsson PA. Pathogenesis of visna. I. Sequential virologic, serologic and pathologic studies. Lab Invest 1976; 35: 402-412.
Querat G, Barban V, Sauze N, Filippi P, Vigne, Russo P, Vitu C. Highly lytic and persistent lentiviruses naturally present in sheep with progressive pneumonia are genetically distinct. J Virol 1984; 52: 672-679.
Lechat E, Milhau N, Brun P, Bellaton C, Greenland T, Mornex JF, Le Jan C. Goat endothelial cells may be infected in vitro by transmigration of caprine arthritis-encephalitis virus infected leucocytes. Vet Immunol Immunopathol 2005; 104: 257-263.
Rea-Boutois A, Pontini, Greenland T, Mehlen P, Chebloune Y, Verdier G, Legras-Lachuer C. Caprine arthritis–encephalitis virus induces apoptosis in infected cells in vitro through the intrinsic pathway. Virology 2008; 375: 452-463
Brassfield AL, Adams DS, Crawford TB, McGuire TC. Ultrastructure of arthritis induced by caprine retrovirus. Arthritis Rheum 1982; 25: 930-936.
Dahlberg JE, Gaskin JK, Perk K. Morphological and immunological comparison of caprine arthritis encephalitis and ovine progressive pneumonia viruses. J Virol 1981; 39: 914-919.
Tantaswasdi U, Wattanavijarn W, Pinyochon W, Malayaman A, Supcharoen A, Pachariyanon S. Caprine arthritis-encephalitis like virus infection in Saanen goats. In: Proceedings of the 12th Annual Veterinary Conference; Bangkok, Thailand; 1985. pp. 376-377.