Establishment of Hybridoma Colonies Secreting Monoclonal Antibodies Specific for Tanapox and Swinepox Early Secretory Proteins
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Authors
Utarnachitt, Richard B.
Issue Date
1996
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Poxviruses employ proteins called "virokines" to help overcome the host's immune
system. Virokines share significant homology with certain cytokine receptors. This
allows virokines to bind to cytokines and prevent their interaction with the native receptors.
Neutralizing this interaction between cytokines and their native receptors helps disable the
host's immune system. These virokines are usually classified as early proteins because
they appear before the replication of the viral genome (Fields and Knipe, 1991). Studying
the biological mechanism of these early proteins would serve to expand our understanding
of how viruses evade the immune system. Tanapox virus (TPV) and swinepox virus
(SPV) are two poxviruses used to study the activity of these proteins.
TPV produces an early secretory protein with a mass of approximately 38 kDa
which binds to IL-2, IL-5 and IFN-y (Essani et al., 1994) Neutralization of these cytokines
would allow TPV to incapacitate the TH I cell mediated and TH2 associated immune
response, crippling specific anti-viral immunity. Spy produces an uncharacterized early
secretory protein with a mass of approximately 34 kDa which may be involved in the
mimicry of IFN-y receptors and G-protein coupled receptors. Interfering with the function
of these proteins would inhibit normal immune responses including antibody production, T
cell function, expression of cell surface antigens, regulation of natural killer cells,
macrophage function and G protein function (Mas sung et al., 1993).
The first step in elucidating the function of these viral secretory proteins is to isolate
pure protein from the media or supernatant surrounding virus infected cells. The
monospecific binding capability of monoclonal antibodies have proven to be useful in
isolation of a variety of antigens. In this experiment, monoclonal antibodies were
generated against the 38 kDa protein produced by SPY and the 34 kDa protein produced
by TPV. Hybridoma were produced by fusion of spleen cells from immunized Balb/c mice
with myeloma cells (p63Ag8.653). The clones were screened for reactivity to viral antigen
using indirect ELISA. The results of these tests were scored visually. Due to time
constraints, only preliminary results were obtained from this study: Three TPV hybridoma
colonies and four SPY hybridoma colonies reacted to viral antigen. Although it could not
be determined whether the reactive colonies were producing antibody which recognized the
two proteins of interest, the hybridoma developed are some of the first monoclonal
antibodies to recognize antigenic determinants for TPV and SPV.
Description
vi, 24 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.