Researchers Apply New Immune Technologies in Search of Mono Vaccine

Researchers Apply New Immune Technologies in Search of Mono Vaccine
  1. Researchers Apply New Immune Technologies in Search of Mono Vaccine
    Researchers are analyzing the genetics of human immune responses to develop the basis of an effective vaccine or drug therapy against Epstein-Barr Virus,…

A micrograph of the axisymmetric flow-focusing nozzle during emulsion generation.Image courtesy of the University of Kansas

LAWRENCE — A research lab at the University of Kansas School of Engineering and School of Pharmacy is analyzing the genetics of human immune responses to develop the basis of an effective vaccine or drug therapy against Epstein-Barr Virus, or EBV, the pathogen that causes mono and infects around 90-percent of adults worldwide.

The virus, which commonly spreads during teen years and young adulthood via kissing, has hit-or-miss health consequences. In some people, EBV causes infectious mononucleosis and can induce cancers like gastric carcinoma or nasopharyngeal carcinoma in later years—while in others the virus is successfully suppressed by the immune system. No EBV vaccine has been approved by the FDA, and the mechanisms of immune protection from EBV-associated diseases are still unclear.

“We are applying recently developed technologies to address an age-old problem,” said Brandon DeKosky, assistant professor of chemical and petroleum engineering and pharmaceutical chemistry at KU, who is leading the effort. “Why do some people infected with Epstein-Barr Virus have a terrible time and get infectious mononucleosis for months, while others are completely asymptomatic and don’t even know they have the virus?”

MOPC-21 immortalized B cellsMOPC-21 immortalized B cells encapsulated in emulsion droplets. The outer aqueous stream that normally contains cell lysis buffer (Fig. 1a, gray solution) was replaced with 0.4% Trypan blue in PBS to examine cell viability throughout the flow focusing and emulsification process. Emulsified cell viability was approximately 90% and cell viability did not differ substantially from non-emulsified controls.Image courtesy of the University of KansasWith a recent $1.8 million, five-year grant from the National Institutes of Health, DeKosky’s lab is using high-throughput techniques to analyze the behavior of B cells in the body. A vital part of our immune system, B cells are white blood cells specialized to bind to specific antigens, such as EBV, and produce antibodies that neutralize foreign agents.

“Each of us has around 100 billion B cells, and each one is playing its own unique role in protecting us from disease,” DeKosky ?said. “My lab looks at single B cells en masse to figure out which one is doing what—and we try to use that molecular-scale information to address larger questions of why one person is protected, why another person might feel sick, and what’s happening with the immune system in both situations.”

The KU researcher first isolates B cells that are reactive to EBV, which is a complicated task due to the sheer numbers of cells involved. DeKosky’s team has honed an innovation that mixes millions of B cells into an emulsion to pinpoint those tailored to fight EBV.

“Instead of taking single B cells and putting one per tube, which is the traditional and more time-consuming method, we put cells into emulsion droplets, which are droplets of water separated from each other by oil—just like salad dressing,” DeKosky said. “Imagine you have a million B cells isolated from a patient, and if you put them in shaken-up salad dressing you now have a million cells isolated into their own individual compartments. Then we take genetic material from each of those single cells in each of those droplets—you can call them micro-reactors. Just one person in one afternoon can screen millions of B cells at the single-cell level using this technology.”

The project will track “longitudinal samples” of B cells, meaning they follow participants’ immune systems over time to see differences across different people, and they also study how a person’s immune system changes as they progress from initial acute infection to long-term immune suppression of the virus.

Related Article: Virus Hydrophobicity Can Help Purify Vaccines

“This class of viruses infects people for life, so it has an initial infectious stage and eventually goes latent but can reactivate in some cases, for example if a person is ever immunocompromised like after a transplant or similar…

A micrograph of the axisymmetric flow-focusing nozzle during emulsion generation
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