Development of monoclonal antibodies against bovine herpesvirus-1 (BHV-1)

Gurpreet Kaur; P N Dwivedi; Ramneek Ramneek; N K Maiti

doi:10.56093/ijans.v82i7.21744

Authors

Gurpreet Kaur Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab141 004 India
P N Dwivedi Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab141 004 India
Ramneek Ramneek Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab141 004 India
N K Maiti Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab141 004 India

https://doi.org/10.56093/ijans.v82i7.21744

Keywords:

AGPT, Bovine Herpesvirus-1, ELISA, Hybridoma cell culture, Monoclonal antibody.

Abstract

The BHV-1 antigen was produced in bulk by propagating BHV-1 in Madin Darby bovine kidney cell line. The antigen purified by 20–40% sucrose density gradient centrifugation was confirmed for presence of virus. The BALB/C mice were immunized by purified virus via intraperitoneal route. The spleenic lymphocytes from immunized mice were fused with myeloma cells using polyethylene glycol as a fusion agent and suspended in HAT (hypoxanthine aminopterin thymidine) medium for differentiation of hybrids. A total of 11 clones were observed, out of which 7 were positive for antibody production. The supernatant of all the positive clones showed very low concentration of antibody, except the clone B6.1, where the concentration of antibody was higher than other positive clones. The culture supernatant of B6.1 hybrid clone was isotyped by enzyme linked immunosorbent assay (ELISA) based isotyping kit and isotype of monoclonal antibody was of IgM class. In western blotting, the IgM monoclonal antibody identified the 27 KD protein of BHV-1.

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References

Babuik L A, L’Italien J, VanDrunen Little-Van den Hurk S, Zamb T, Lawman M J, Hughes G and Gifford G A. 1987. Protection of cattle from bovine herpesvirus type-1 (BHV-1) infection by immunization with individual viral glycoproteins. Virology 159: 57–66.

Bolton D C, Zee Y C and Ardaus A A.1983. Identification of envelope and nucleocapsid proteins of infectious bovine rhinotracheitis virus by SDS-polyacrylamide gel electrophoresis. Veterinary Microbiology 8: 57–68.

Carter P B, Beegle K H and Gebhard D H. 1986. Monoclonal antibodies: critical uses and potential. Veterinary Clinics of North American Small Animal Practice 16: 1171–79.

Collins J K, Butchner A C and Reigel C A. 1984. Neutralising determinants defined by monoclonal antibodies on polypeptides specified by bovine herpesvirus-1. Journal of Virology 52: 403– 09.

Collins J K, Butchner A C and Reigel C A. 1985. Immune response to bovine herpesvirus type-1 infections: virus-specific antibodies in sera from infected animals. Journal of Clinical Microbiology 21: 546–52.

Durhams P J K, Hassard L E and Donkersgoed J Van. 1991. Serological studies of infectious bovine rhinotracheitis, parinfluenza-3, bovine viral diarrhea and bovine respiratory syncitial virus in calves following entry to a bull test station. Canadian Veterinary Journal 32: 427–29.

Florent C and Demarneffe C. 1986. Enzyme linked immunosorbent assay used to monitor serum antibodies to bovine respiratory disease viruses. Veterinary Microbiology 11: 309–14.

Gibbs EPJ and Rweyemamu M M. 1977. Bovine herpesvirus-1, Bovine Herpesvirus-1. Veterinary Bulletin 47: 317–43.

Gregersen J P, Pauli G and Ludwig H. 1985. Bovine herpesvirus-1. Differentiation of IBR and IPV viruses and identification and functional role of their major immunogenic components. Archives of Virology 84: 91–103.

Kida H, Brown L E and Webster R G. 1982. Biological activity of monoclonal antibodies to operationally defined antigenic regions on the haemagglutinin molecule of A/Seal?Massachusetts/1/80 (H7 N7) influenza virus. Immunology 122: 38–47.

Laemmli U K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Nature 227: 680–85. Lowry O H, Rosebrough N J, Farr A L and Randall R J. 1951.

Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry 193: 265–75.

Marshall R L, Rodriguez L L and Letchworth G J. 1986. Characterisation of envelope proteins of infectious bovine rhinotracheitis virus (bovine herpesvirus-1) by biochemical and immunological methods. Journal of Virology 57: 745.

Marshall R L, Israel B A and Letchworth III G J. 1988. Monoclonal antibody analysis of bovine herpesvirus-1 glycoprotein antigenic areas relevant to natural infection. Virology 165: 338–47.

Mayfield J E, Good P J, Van Oort H J, Campbell A R and Reed D E. 1983. Cloning and cleavage site mapping of DNA from BHV- 1 (Cooper strain). Journal of Virology 47: 259–64.

Mehrotra M L, Rajya B S and Kumar S. 1976. Infectious bovine rhinotracheitis (IBR), Keratoconjunctivitis in calves. Indian Journal of Veterinary Pathology 11: 70–72.

Misra V, Blumenthal R M and Babiuk L A. 1981. Proteins specified by bovine herpesvirus type-1 (infectious bovine rhinotracheitis virus). Journal of Virology 40: 367–78.

Mohanty S K, Kataria J M, Swain P and Verma K C. 2000. Development of hybridoma secreting monoclonal antibodies against a velogenic New Castle disease virus isolated from guinea fowl (Numida meleagris). Indian Journal of Comparative Microbiology Immunology and Infectious Disease 21 (1): 63–65.

Sreenivasa B P, Natrajan C, Gajendragad M R, Suryanarayana V V S and Rasool T J. 1998. Polypeptide analysis of Indian isolate of Bovine Herpesvirus-1 (BHV-1). The Indian Journal of Animal Sciences 68 (2): 230–33.

Srikumaran S, Onisk D V, Borca M V, Nataraj C and Zamb T J. 1990. Anti-idiotypic antibodies induce neutralizing antibodies to bovine herpesvirus-1. Immunology 70: 284–89.

Studdert M Z. 1989. Bovine encephalitis herpesvirus. Veterinary Record 125: 584.

Suresh K B, Sudarshan K J H and Rajasekhar M. 1999. Seroprevalence of infectious bovine rhinotracheitis in India. Indian Veterinary Journal 76: 5–9.

Van Drunen Little-VanDen Hurk S, VanDen Hurk J V, Gilchrist J E, Misra V and Babuik L A. 1984. Interactions of monoclonal antibodies and bovine herpesvirus type-1 (BHV-1) glycoproteins: Characterization of their biochemical and immunological properties. Virology 135: 466–79.

Van Drunen Little-VanDen Hurk S and Babiuk L A. 1986. Polypeptide specificity of the antibody response after primary and recurrent infection with bovine herpesvirus-1. Journal of Clinical Microbiology 23: 274–82.

Pastoret P P, Jelleur P, Aguilar-Setien A, Leroy P, Schwers A, Godart M and Schoenaers F. 1979. Effect de la dexamethasone sur le titre et la taille des plages du virus de la rhinotracheite infectieuse bovine (Bovid Herpesvirus 1, IBR). Annales de Médecine Vétérinaire 123: 397–402.