Comparing Antibody-Coated Immune Beads with Flow Cytometry to Measure ß-2-Microglubulin+ Murine Spleen Cells
Nancy Bigley (Advisor), Barbara Hull (Committee Member), Dawn Wooley (Committee Member)
Master of Science (MS)
In diseases like AIDS, Multiple Myeloma, Multiple Sclerosis (MS), Type 1 Diabetes, and Systemic Lupus Erythromatosus the detection of major histocompatibility complex type 1 molecules (MHC 1) can be a helpful component of disease diagnosis and prognosis. Most somatic mammalian cells display to varying degrees major histocompatibility complex (MHC) class 1 antigen on the cell surface. MHC1 molecules consist of a polymorphic alpha a chain and a monomorphic beta chain, beta 2 microglubulin (ß2m). The ß2M composition is fairly constant within a species whereas the alpha chain is not. The polymorphic alpha chain is encoded by an MHC complex gene that is specific for each member of the species. The consistency of the ß2M protein within a species makes it a superior target to the variable a chains when attempting to quantify MHC1 molecules. Currently, there are no reagents available to identify the MHC1 antigen within an outbred population such as human or laboratory animals other than defined in bred mouse strains. In this study, the presence of MHC1 molecules on spleen cells from an outbred mouse strain (ICR strain) was evaluated by looking for monomorphic ß2m molecules. The two methods, which were used to obtain the numbers of ß2m+ mouse spleen cells, were flow cytometry and the antibody-latex bead method. Similar numbers of ß2m- positive cells were obtained using these two methods (7-9%). These results demonstrate that the antibody-coated beads are a suitable, less expensive and time saving alternative to determine the numbers of ß2m+ positive cells when compared to flow cytometry.
Department or Program
Microbiology and Immunology
Year Degree Awarded
Copyright 2016, all rights reserved. My ETD will be available under the "Fair Use" terms of copyright law.