Production and Characterization of Respiratory Syncytial Virus Specific T Cell Hybridomas Using FG, a Novel Chimeric Glycoprotein
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Authors
Pomaranski, Mark R.
Issue Date
1989
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Respiratory syncytial virus has expressed on its surface two
glycoproteins, F and G, which are considered to be primary targets
for host immune responses. Through the use of a baculovirus
expression vector system, a novel chimeric glycoprotein, FG, which
contains the extracellular regions of both F and G glycoproteins, has
been expressed in an insect derived cell line. To gain a more comprehensive
understanding of the role of glycoprotein presentation in T cell
stimulation, multiple T cell hybridomas specific for the chimeric FG
glycoprotein were produced. Initially popliteal lymph node cells
(PLNC) from BALB/c mice that had been immunized with FG were
restimulated in vitro to determine optimal time and concentration of
FG for maximum responsiveness to the glycoprotein. PLNC were then
fused with BW5147 thymoma cells using polyethylene glycol (PEG).
Resulting T cell hyridomas were then screened for specific reactivity
to FG by measuring the release of IL-2 after stimulation with FG and
appropriate antigen presenting cells (APC). Approximately 4% of the
cell fusion hybridomas reacted specifically with FG, one of which
(106) was chosen for a detailed characterization owing to its stability.
Using monoclonal antibody (MAb) blocking and mouse L cells
transfected with class II Major Histocompatibility Complex (MHC)
genes, the hybridoma 106 was tested to determine which class II
MHC molecules were presenting FG. These characterization studies
have shown that the lAd determinant is the class II MHC molecule
responsible for FG presentation to this particular hybridoma. The
continued testing of this hybridoma will allow more complete
characterization and mapping of immunological sites on the two
major glycoproteins of RS virus.
Description
vii, 31 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.