Investigating the Role of Fc?Rlla on Neutrophil Ability to Perform Trogocytosis on Trichomonas vaginalis

Trichomoniasis is a sexually transmitted infection caused by a protozoan pathogen named Trichomonas vaginalis (Tv). Tv affects more than 300 million men and women worldwide. It has been shown that neutrophils use a process called trogocytosis to kill Tv. Trogocytosis is a process in which a cell, such as a neutrophil, takes "bites" out of its neighboring target cell. It has been shown that the neutrophil trogocytosis may be regulated by antibody opsonization of its target cell. Antibody opsonization is the coating of a pathogen with antibodies to crosslink cells to the pathogen using Fe surface receptors. Research has shown that Fc?Rlla (CD32a) is important for the trogocytosis mechanism against cancer cells. We hypothesize that CD32a is also important for neutrophils to trogocytose and kill Tv. To test this, we will knock out the CD32a gene using the CRISPR/Cas9 system. We will generate CD32a knockout neutrophil-like cells (NLCs) and perform several assays that will show whether trogocytosis was affected by that CD32a knock-out. We have generated a plasmid that will mediate CD32a knockout and confirmed the presence of CD32 in NLCs through antibody staining and flow cytometry. We will also perform cytolysis assays under different inhibiting and blocking conditions of the CD32a receptor to further investigate the role of CD32a in neutrophil trogocytosis of Tv. We hope that our findings will give us more insight into the initiation process of trogocytosis of Tv by neutrophils that can inform prevention treatments in the future.


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