Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Room: Exhibit Hall
Ambria Manuel - University of Missouri - Columbia
Co-Author(s): Deborah Anderson, University of Missouri, Columbia, MO
SARS-CoV-2 is the causative pathogen of Coronavirus Disease 2019 (COVID-19). The severity of COVID-19 varies and is determined, in part, by the immune response of the body. The immune response is primarily controlled by neutrophils and macrophages. Studies show that the human body will produce a high number of neutrophils and macrophages when infected with SARS-CoV-2. Typically, the human body produces immature immune cells when presented with an infection. Neutrophils and macrophages are recruited to the site of infection; the recruitment will primarily happen in organs like the lungs, spleen, liver, and brain. The recruited neutrophils that have infiltrated the lungs break down the inner lining of the lungs. To further understand whereby the virus affects the cells within the human body, an effective mouse model is required to evaluate the severity of the disease. Within the humanized mouse model that expresses the human ACE-2 receptor, the disease has been exhibited to impact the lungs producing additional immature immunosuppressive neutrophils. We have developed an immunohistochemistry assay, in which formalin-fixed tissues are stained with antibodies CD68 (macrophages) and myeloperoxidase (neutrophils). The antibodies assist in assessing the recruitment of the immune cells within the liver, spleen, and lungs following infection. To comprehend if the stain is effective, mice infected with Yersinia pestis bacteria were utilized to assess the functionality of the staining procedure. We have developed the protocol for sectioning and staining tissues. Our findings indicate that infection results in an increased number of immature neutrophils and macrophages, particularly abnormally shaped. In addition to the lungs, infected spleen and liver were examined. Infection of the cells is determined by the affectivity of the staining, observing the morphology of the macrophages and neutrophils. In future studies involving immunohistochemistry, the implementation of the staining protocol is intended to be used on COVID-19-infected tissues. The histology slides will be evaluated based on infectivity, collected in a time-course experiment will be quantified for the recruitment of neutrophils and macrophages to disease lesions.
Funder Acknowledgement(s): NIHMARC/IMSD
Faculty Advisor: Deborah Anderson, firstname.lastname@example.org
Role: I solely did the entirety of this experiment with the help of my PI. So I would do the cutting of the mouse infected tissue and put it on histology slides so that they would be stained. I created an effective staining protocol that took multiple trials to get right and I also analyze the slides after the staining.