Infectious center assay of intracellular virus and infective virus titer for equine mononuclear cells infected in vivo and in vitro with equine herpesviruses.
Abstract: A novel, simple method of infectious center assay was developed to detect and quantitate the intracellular existence of equine herpesvirus 1 and equine herpesvirus 2 in peripheral blood mononuclear cells infected in vivo and in vitro with the viruses by cocultivation of these cells with a permissive equine cell culture. The infectious center titers were correlated with the infectious virus titers. In vivo equine herpesvirus 1-infected mononuclear cells obtained from ponies experimentally infected with the virus and equine herpesvirus 2-infected mononuclear cells obtained from selected naturally infected ponies with the virus gave by infectious center assay a mean value of 67 infectious center/2 x 10(6) cells as a peak titer on day 4 postinfection and 26 infectious center/2 x 10(6) cells for equine herpesvirus 1 and equine herpesvirus 2 respectively. The mononuclear cells, in both cases, did not contain detectable infectious virus, but the infectious virus was detected from the respective cells when they were cultured in the presence of mitogen. The equine herpesvirus 1 infected mononuclear cells in culture gave a mean count of 8.05 x 10(2) infectious center/2 x 10(6) cells/mL and contained 1.08 x 10(4) plague assay/mL of infectious virus. Similarly the equine herpesvirus 2 infected mononuclear cells in culture gave a mean count of 7.1 x 10(1) infectious center/2 x 10(6) cells/mL and contained <10(1) tissue culture infective dose(50)/mL of infectious virus. Mononuclear cells infected in vitro with equine herpesvirus 1 gave a mean count of 9.3 x 10(4) infectious center/2 x 10(6) cells/mL and contained 5.75 x 10(3) plaque assay/mL of infectius virus. Culturing these cells in the presence of mitogen gave a mean count of 5.5 x 10(3) infectious center/2 x 10(6) cells/mL and contained 9 x 10(3) plague assay/mL of infectious virus. A correlation between infectious center assay and infectious virus assay is discussed.
Publication Date: 1983-01-01 PubMed ID: 6299486PubMed Central: PMC1235886
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- Comparative Study
- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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The paper describes the development of a new, simple method of infectious center assay to detect and quantify equine herpesviruses 1 and 2 in horse blood cells. The researchers used these tests to assess cells infected in the lab and in live horses, in order to establish a correlation between the number of infected cells and the total infectious virus levels.
Research Methods & Observations
- The paper introduces a novel and simplified method of infectious center assay, a laboratory technique used for identifying and determining the amount of equine herpesvirus 1 and 2 inside the horse’s peripheral blood mononuclear cells.
- The cells for study were infected with the viruses either in a natural environment (in vivo) or in a lab setting (in vitro).
- The study focused on calculating infectious center titers (a measure of viral concentration) and comparing it with infectious virus titers (more direct count of the total infective viral particles).
- In the in vivo experiment, mononuclear cells obtained from ponies infected with equine herpesvirus 1 and 2 showed infectious center peaks four days post-infection. While these cells didn’t show any signs of infective virus, the virus was detectable when cells were grown in a culture in the presence of a mitogen, a substance that promotes cell division.
Findings & Conclusions
- Similarly, in the in vitro experiment, cells infected with the two viruses showed the presence of the virus when cultured with a mitogen.
- The in vitro-infected cells showed a higher count of infectious centers than the in vivo-infected cells.
- In all the scenarios, the virus was only detectable when the cells were cultured with a mitogen.
- A potential correlation between the infectious center assay and the infectious virus assay – implying that the number of cells harboring the virus is proportional to the total infectious virus levels – is suggested and is a topic for further discussion.
Cite This Article
APA
Dutta SK, Myrup AC.
(1983).
Infectious center assay of intracellular virus and infective virus titer for equine mononuclear cells infected in vivo and in vitro with equine herpesviruses.
Can J Comp Med, 47(1), 64-69.
Publication
Researcher Affiliations
MeSH Terms
- Animals
- Herpesviridae / isolation & purification
- Herpesviridae / pathogenicity
- Herpesviridae Infections / microbiology
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / isolation & purification
- Herpesvirus 1, Equid / pathogenicity
- Horse Diseases / microbiology
- Horses
- In Vitro Techniques
- Monocytes / microbiology
- Perissodactyla
- Virus Cultivation / methods
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Citations
This article has been cited 8 times.- Desai D, Londhe R, Chandane M, Kulkarni S. Altered HIV-1 Viral Copy Number and Gene Expression Profiles of Peripheral (CEM CCR5+) and Mucosal (A3R5.7) T Cell Lines Co-Infected with HSV-2 In Vitro. Viruses 2022 Aug 4;14(8).
- Jin YH, Kim CX, Huang J, Kim BS. Infection and Activation of B Cells by Theiler's Murine Encephalomyelitis Virus (TMEV) Leads to Autoantibody Production in an Infectious Model of Multiple Sclerosis. Cells 2020 Jul 27;9(8).
- Oladunni FS, Horohov DW, Chambers TM. EHV-1: A Constant Threat to the Horse Industry. Front Microbiol 2019;10:2668.
- Yoshikawa T, Hill TE, Yoshikawa N, Popov VL, Galindo CL, Garner HR, Peters CJ, Tseng CT. Dynamic innate immune responses of human bronchial epithelial cells to severe acute respiratory syndrome-associated coronavirus infection. PLoS One 2010 Jan 15;5(1):e8729.
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- Giessler KS, Goehring LS, Jacob SI, Davis A, Esser MM, Lee Y, Zarski LM, Weber PSD, Hussey GS. Impact of the host immune response on the development of equine herpesvirus myeloencephalopathy in horses. J Gen Virol 2024 May;105(5).
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