Feline immunodeficiency viruses (FIVs) are naturally occurring lentiviruses (family Retroviridae) of felid species, including domestic and wild cats. Studies on FIVs are beneficial for understanding the host immune response associated with disease progression (e.g., domestic cat FIV) or the viral kinetics and molecular ecology associated with naturally occurring infections in wildlife (e.g., bobcat and mountain lion FIVs). Here we describe the development and validation of the following microsphere immunoassays (MIAs) for evaluating the cytokine and antibody response of domestic cats: i) the quantification of cytokines (interferon gamma (IFNγ), interleukin (IL)-10, and IL-12/IL-23) in cell culture supernatant, and ii) the quantification of these cytokines in plasma; iii) the quantification of total IgG and IgA in plasma, and iv) the detection of IgG and IgA antibodies to feline CD134 (the primary cell receptor for FIV), and FIV capsid (CA) and surface (SU) proteins in plasma. These assays were used to evaluate temporal cytokine and antibody responses of domestic cats experimentally infected with various FIV strains. To analyze viral RNA loads associated with naturally occurring FIV infections in bobcats or mountain lions, we are adapting existing quantitative PCR assays for use with plasma samples. The eight assays described here are/will be beneficial for addressing questions related to lentiviral immune response and viral kinetics.
|Commitee:||Avery, Paul, Hussey, Gisela, Zabel, Mark|
|School:||Colorado State University|
|Department:||Microbiology, Immunology, & Pathology|
|School Location:||United States -- Colorado|
|Source:||DAI-B 74/10(E), Dissertation Abstracts International|
|Subjects:||Microbiology, Virology, Immunology|
|Keywords:||Antibodies, Cytokines, Fiv/plv, Microsphere immunoassays, Plasma viremia|
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