Ayush Kumar

 

My name is Ayush Kumar and I was born in the Fiji Islands and immigrated to the United States when I was five to the state of Oregon. In order to broaden my scope of awareness in research, I applied and was accepted to the NSF REU “Exploring the Anthropocene” at the Great Rivers Field Station and received mentorship from Dr. Colaninno-Meeks and Dr. Chick. The goal of the research was to use ecological and archaeological datasets to understand how anthropogenic factors affect water quality and fish communities. I spent the majority of my time on large fishing boats on the Mississippi River electrofishing and collecting water quality samples. My goal was to analyze and relate species richness to various chemical concentrations in locations along the river. Ultimately, we wanted to gauge how population distribution changed over time due to pollution, hydrologic dams, and levees. This experience was formative for me because I gained experience with population health and wildlife management and also field and laboratory work, including working with live wild organisms and performing quantitative analysis with large data sets. I was also able to present my research at the Mississippi River Research Consortium on how water quality has affected fish communities since the Clean Water Act, further developing my public speaking and communication skills. As a biology major at a small liberal arts college, I had limited options for research on campus. To gain research experience, I interned at the Oregon National Primate Center (ONPRC) in the Vaccine and Gene Therapy Institute (VGTI) where I developed my technical skills and virology knowledge. Working alongside postdoctoral associate, Dr. Travis Whitmer, I learned about HIV’s ability to evade detection by T-Helper Cells, which typically detect invaders using their MHC Class I proteins. The main goal of the project was to use HCMV as a vector to express foreign antigens (HIV/TB), which generates novel T cell responses that are protective against their corresponding diseases. We were able to look at the closely related rhesus macaque CMV and test these different constructs in vivo to see how the primate immune system responds. Thus, this would ideally provide us with an idea of how a human test subject would respond to HCMV. I was responsible for the molecular portion of the project: cloning, conducting RT PCR, examining DNA segments virtually, and checking the accuracy of the clones for further use in future experiments. My role in the project allowed me to develop technical skills in sequencing and cloning. As a result, I became interested in virology and disease ecology due to my mentorship with Dr. Whitmer, which further initiated my desire for graduate school. Education: Concordia University, Honors Program, Major: Biology, Minor(s): Psychology and Spanish Description: In August 2018, I started working at the UGA Poultry Diagnostic and Research Center in the Rajao/Perez Lab. In the beginning I was trained in basic lab techniques and I was able to work with a graduate student, Brittany Seibert and my mentor Dr. Cardenas Garcia. With the exceptional mentorship, I was able expand my knowledge about influenza A and B and learn techniques associated with Reverse Genetics and vaccine development. Influenza A (IAV) and Influenza B (IBV) are enveloped viruses with a negative-sense, segmented, single-stranded RNA genome, with eight viral RNA (vRNA) segments. For my project, I primarily worked with influenza B, which can cause outbreaks of seasonal flu and can be just as severe as Influenza A. Influenza B was only known to infect humans (mainly adolescents, school children, and geriatric patients) and comprises up to 20-30% of infections. Influenza B also evolves slower than Influenza A viruses. It has been reported that this virus can infect pigs (Ran, et al., 2015).
Research: In this project, I worked with Influenza B/Brisbane, which was isolated in Australia in 2008. So the major goal is to evaluate to which extent to use swine polymerase I promoter (spolI) with the B/Bris virus. Why do we want to test the spoll? The reason we want to test this is because pol I promoters are species specific and we want to determine if this reverse genetics systems carrying the spoll promoters will be more efficient than hpolI. We also want to assess the viability of a reverse genetics vector carrying the spolI promoter to rescue recombinant IBVs in swine and human origin cells.
We investigated the viability of reverse genetics plasmid vectors containing the swine pol I promoter (spoll) and B/Brisbane/60/2008 (B/Bris) influenza virus, and their capability for virus rescue in HEK293T/MDCK or PK-15/MDCK co-cultures. These viruses are derived from full length cDNA of the influenza viral gene segments. Therefore, with the ability to generate recombinant viruses that have mutations in the viral genome, we can identify the viral and host factors that contribute to influenza pathogenesis, transmissibility, host-range, host/pathogen interactions and restrictions, and virulence. First, we subcloned all eight gene segments from B/Bris into a reverse genetics vector containing the spoll, giving origin to eight plasmids, each one containing a gene segment. Then, we assessed the viability of the newly generated plasmids by transfecting HEK293T/MDCK co-cultures using the 7+1 method (seven plasmids known to function properly and one plasmid containing the gene of interest and the spoll). The project allowed us to understand the potential of spoll for in vivo IV reverse genetics in swine models.
Future: After completing the PREP@UGA program, I will be attending Ontario Veterinary College at the University of Guelph Doctor of Veterinary Medicine Program. My plan after receiving my DVM is to continue on in a PhD program and pursue my interest in Laboratory Animal Medicine. For me PREP@UGA was a steppingstone towards gaining experience in research and understanding the expectations that graduate schools have. Thus, PREP@UGA provided me with exceptional research opportunities, mentorship, and support for my future.