This summer I have the pleasure of working for the University of Hawaii John A. Burns School of Medicine Hawaii Center for AIDS located in downtown Honolulu. I am working in Dr. Bruce Shiramizu’s lab where we focus on clinical research with the aim to contribute to the advancement of HIV treatment and extension of patients’ lifespan. Based on the ongoing projects, the lab staff assisted me with creating a specific project to work on over the course of the summer. We decided that my project would investigate the effect of blood plasma on the integrity of an in vitro Blood-brain barrier (BBB).
In the past, a diagnosis of HIV was more or less a death sentence. Through educating at-risk groups and advancing technology, HIV became better managed at an epidemiological level. Even further, therapy today is very effective and patients are able to live longer lives. cART or combination antiretroviral therapy utilizes a number of different classes of drugs to thwart HIV proliferation throughout the body. However, the virus is not completely eradicated from the body because a reservoir of HIV remains hidden in host DNA within white blood cells. If ever activated, the virus can be spread throughout the body once again causing a number of negative consequences.
HIV is very harmful to many different organ systems within the body and having a reservoir of HIV can be harmful to the brain. There is a protective layer called the BBB that acts to protect the brain to keep what belongs in the brain in and what does not belong out of the brain. However, blood cells have the ability to cross through the BBB. This means that even HIV infected white blood cells can enter the brain and circumvent the protective measure of having a BBB. When infected blood cells get in the brain, the integrity of the BBB is disrupted ultimately causing a decrease in cognitive function.
As my project is focused on the BBB, some of my responsibilities include growing and maintaining cell cultures to be utilized in making the BBB and then keeping the barrier alive as well. Cell culturing is essential to my greater project to ensure viable endothelial cells and astrocytes are available for growing barriers. After a week, a number of barriers are used to measure how confluent the barrier is with a probe (TEER: transepithelial electric resistance) and then how “leaky” it is with a blue dye (EBA: Evan’s blue dye albumin conjugate). The rest of the barriers are used to perform the actual experiment where white blood cells and plasma (no plasma, HIV positive, HIV negative) are placed in an insert above the barrier to measure white blood cell transmigration through the barrier. In other words, if the barrier is not very confluent and very leaky, the white blood cells will pass through the barrier very easily. Research shows that when HIV is present even more white blood cells can get through. Therefore we predict that if HIV positive plasma is added to white blood cells, then there will be a decrease of barrier integrity determined by cell counts, TEER, EBA. But for now, this remains TBD.
Working in the lab has been a great learning opportunity and I have also been able to learn about HIV and meet other undergraduate students in the summer program. Once a week, we come together to have lunch and listen to a presentation given by one of the principal investigators from the Hawaii Center for AIDS (HICFA). The presentations provide supplementary information on HIV that contributes to enhancing our overall understanding of the virus and also provides a space to think critically and ask questions.
Conducting research at JABSOM has definitely been the highlight of my summer and it has been an amazing experience to meet and interact with students and professionals in this field. I am very thankful to be given independence in the lab and the responsibility of having my own project. My work in the lab pushes me to apply my biology background and develop more confidence in a hands-on environment—truly an invaluable experience.
Experiences like Kai’s are made possible by the Whitman Internship Grant, which provides funding for students to participate in unpaid internships at both for-profit and non-profit organizations. To learn how you could secure a Whitman Internship Grant or host a Whitman intern at your organization, click here or contact Assistant Director for Internship Programs Victoria Wolff.