Making a Full Circle back to ETSU
Center of Excellence in Inflammation, Infectious Disease and Immunity (CIIDI)
I began my career at Quillen College of Medicine and came full circle back to East Tennessee State University. I am currently a Clinical Laboratory Technician in the Department of Surgery. Our lab is affiliated with the ETSU Center of Excellence in Inflammation, Infectious Disease and Immunity (CIIDI). We study sepsis and aging of the immune system. I no longer work with laboratory animals.
My boss is a Doctor of Veterinary Medicine and has a Ph.D. in Biomedical Sciences. She is interested in the cellular and molecular mechanisms of sepsis as well as innate immune recognition of the fungal cell wall. We are currently investigating Candida albicans. Our main focus is to better understand why the healthy aging population and the septic patient are more vulnerable to fungal infection.
Candida albicans is the most common human fungal pathogen. It is normally a harmless commensal organism. However, it is a opportunistic pathogen for some immunologically weak and immunocompromised people. It is responsible for painful mucosal infections such as the vaginitis in women and oral-pharyngeal thrush in AIDS patients. In certain groups of vulnerable patients it causes severe, life-threatening bloodstream infections and subsequent infections in the internal organs. - Candida albicans, a major human fungal pathogen
Donors and Patients
We get whole blood from young (18-29), middle-aged (30-59), and elderly (60+) healthy donors, sepsis patients from the hospital and infectious disease patients from the clinic. We isolate immune cells from the blood. Neutrophils are isolated using immunomagnetic negative selection. Peripheral blood mononuclear cells (PBMCs) are isolated using a special CPT tube containing sodium citrate and Ficoll Hypaque solution.
Neutrophils are the most abundant type of white blood cells. In healthy adults, they typically constitute about 50 to 70 percent of white blood cells and function as the first line of defense against bacteria and other foreign organisms. - Neutrophils
Neutrophils are white blood cells with a multilobed nucleus
Human peripheral blood mononuclear cells (PBMCs) are isolated from peripheral blood and identified as any blood cell with a round nucleus (i.e. lymphocytes, monocytes, natural killer cells (NK cells) or dendritic cells). - Peripheral Blood Mononuclear Cells
Experiments
We perform several experiments on the isolated immune cells. We stain the cells with fluorescent antibodies to detect pattern recognition receptors on the cell surface by flow cytometry. There is a test to determine uptake efficiency of Candida albicans by neutrophils using flow cytometry as well. We also look at production of reactive oxygen species (ROS) by neutrophils.
We compare several cytokines produced by PBMCs. Cytokines are chemical messages that immune cells use to talk to each other. We test the growth media from PBMCs for these cytokines by enzyme-linked immunosorbent assay (ELISA).
We manage the Flow Cytometer for the college.
BD LSRFortessa X-20 Cell Analyzer at ETSU - Flow Cytometry Core Facility
photo taken with my cellphone
HL-60 Transfection Project
I am currently working to develop a stable transfection of HL-60 (Human Leukemia) cell line to overexpress a pattern recognition receptor that recognizes fungal elements. This protein is upregulated on septic neutrophils. Once I create a stable transfection, I will test these cells the same way I have tested the immune cells of healthy donors, sepsis and infectious disease patients. HL-60s are neutrophil-like in nature. These transfected HL-60 cells will be used to model septic neutrophils.
Summer Research Program at ETSU-QCOM
Each summer, several research labs take one or two medical students as part of a grant program to expose them to biomedical research. These students follow us around for 6-8 weeks to learn all about what we do in the lab. In 2019, we had two students who helped us design the ROS experiments. In 2020, students weren't allowed in the research labs, but we did have one student that learned about our research over Zoom. In 2021, we had one student, and she is my favorite so far. She helped me get all of my data organized for analysis. All the students were very eager to learn. I'm excited to have another medical student this summer!
This concludes my journey through biomedical research. I will probably retire from ETSU. For now, I'm just enjoying being back home in Tennessee. Eureka!
Peace be with you!!!
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