Publication Date
2015
Document Type
Thesis
Committee Members
Nancy Bigley (Committee Member), Cheryl Conley (Committee Member), Barbara Hull (Advisor)
Degree Name
Master of Science (MS)
Abstract
Major histocompatibility complex-Ι (MHC Ι) is an essential protein located on the plasma membrane of every nucleated cell; it contributes to the immune system's ability to defend against bacteria and virus. MHC Ι displays foreign antigens on the cell surface in a manner that encourages CD8+ T cytotoxic cells (CTLs) to respond against these antigens by killing the infected cells. MHC Ι is composed of two chains: a light chain (β-macroglobulin; β2m) and a heavy chain (α chain with 3 domains). The α chain genes have different haplotypes, dependent on the strain of mouse. H-2Kb/ H-2Db is the gene for the α chain in C57 BL/6 mouse.
In this study, we aimed to determine the expression level of MHC Ι in different types of tissues. Since the antibodies for detecting MHC Ι α chains are very expensive, the main goal of the study was to detect both MHC Ι chains in C57 BL/6 healthy mouse tissues and tissues from a mouse infected with parvovirus by using a less-expensive antibody against the β2m chain instead of a monoclonal antibody against the α chain. Moreover, the study aimed to determine the amount of MHC Ι normally expressed in different tissues of C57 BL/6 mouse using immunohistochemistry of paraffin (IHC-P) tissue, with the goal of identifying the organs / tissues that express MHC Ι most abundantly. The analyses of data were performed by using One-way ANOVA.
We found that the expression of both MHC Ι chains in different mouse tissues (spleen, stomach, small and large intestine) was equivalent and the statistical analysis showed that there was no significant difference between the expression of both antibodies (H2Kb/H-2Db and β2m) in healthy tissues or in tissue infected with MPV-1. Additionally, we observed that the highest MHC Ι expression level to be in the small intestine, with the next highest abundance in the large intestine, followed by the stomach, and spleen compared to other healthy tissues (brain, kidney, liver, and heart). Moreover, expression of neither MHC Ι chain could be detected in certain healthy mouse tissue (brain, kidney, liver, and heart). In general, the intensity of 3,3'-Diaminobenzidine (DAB) (brown coloration) of both MHC Ι chains was less in healthy tissues in comparison to the tissues infected with MPV-1, suggesting that healthy tissue expresses low levels of MHC Ι.
Page Count
67
Department or Program
Department of Biological Sciences
Year Degree Awarded
2015
Copyright
Copyright 2015, some rights reserved. My ETD may be copied and distributed only for non-commercial purposes and may not be modified. All use must give me credit as the original author.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
ORCID ID
http://orcid.org/0000-0003-4973-7209
