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Cytomegalovirus (CMV) is a fairly ubiquit ous virus that is capable of remaining unseen by the body’s immune system until the imm une system is compromised. The lack of immune response to CMV-infected cells is due to the virus’ ability to interfere with MHC class I and II. Cytotoxic T Lymphocytes (CTLs) kill cells presenting cognate peptide presented on MHC class I using pe rforin or granzyme . Though it is understood that these work as a part of the body’s ac quired immune system and respond to specific peptides presented on major histocompatibility complexes, little is understood about the exact mechanisms by which CTLs are able to ki ll virus-infected cells. In fact most studies done on CTL killing have been done on peptid e loaded, non –virus-infected cells. In order to explore the mechanisms of CTL killing of infected cells in vitro , we attempted to recreate and expand upon the resu lts of previous lab resear ch involving virus-infected mouse fibroblasts with a wild type (WT) and mutant strain of cytomegalovirus. In the mutant strain of CMV (TKO), we knocked out the three immune evasion genes that interfere with MHC class I presentation sp ecifically. Chromium release assays were performed using T-cells harvested from muri ne OT1 cells lines. Using the original protocol, initial results indicated simila r levels of killing among WT, TKO, and uninfected cells. When the protocol was alte red and T-cells were grown in different cytokine conditions it was shown that cytoki ne IL12 had a particularly strong effect on CTL killing efficacy. Additional chromium releas e assays will determine whether killing of all cells in increased when CTLs ar e grown in IL12. Unde rstanding the ideal environment for CTLs to kill target cells ma y provide a deeper understanding of what is important for an effective immune response.
Mice – Experiments, In Vitro techniques, Cytotoxic T-Lymphocytes, T cells, Cytomegalovirus infections, MHC Class I Genes, MHC Class II Genes, Fibroblasts, Spleen – Immunology, Mice as laboratory animals, Whitman College -- Dissertation collection 2011 -- Biology Department
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