Case study: Solving an assay challenge at Trinity College

When PhD candidate Jeremy Aboagye began working in Professor Ed Lavelle’s Adjuvant Research Lab at Trinity College Dublin, he faced a difficult question: how to accurately measure the mucosal immune response needed for a vaccine against Burkholderia pseudomallei (Bp) – the soil-dwelling bacterium responsible for the often-fatal disease melioidosis.

Jeremy’s project aimed to design a vaccine by combining a Bp antigen with novel adjuvants and delivering it through different immunization routes. His current study focused on a “prime-pull” strategy: first immunizing mice parenterally, then giving an intranasal boost to “pull” protective T and B cells to the mucosa, where they can block infection at the point of entry.

But tracking B cells that secrete antigen-specific IgA – an essential marker for mucosal immunity – proved tricky. Traditional assays like flow cytometry (FACS) require large cell numbers and often involve tagging or modifying antigens. Jeremy needed a method that could handle scarce lung samples while preserving the antigen in its native form.

That’s where Mabtech’s IRIS reader and IgA ELISpot (HRP) Flex kit came in. Using the antigen-specific protocol, Jeremy could enumerate IgA-positive B cells from single-cell lung suspensions with a fraction of the cells FACS would require. Better yet, IRIS’s Relative Spot Volume (RSV) function gave him an estimate of the antibody amount each cell produced.

“The ability to use native antigen and work with ten times fewer cells was a game changer,” Jeremy explained. “The IRIS reader’s RSV data also very nicely complemented ELISAs done on respiratory mucosal samples.”

Earlier this year, Jeremy presented his findings at the Elsevier Vaccine conference in Kyoto, and right now he’s in the midst of finalizing his paper on the results. As for the future, Jeremy is already eyeing FluoroSpot to analyze pulmonary cytokine responses in upcoming experiments. His current findings are paving the way toward a viable Bp vaccine, and his next step – challenging vaccinated mice with the pathogen – will test the protection this novel adjuvant and vaccination strategy can deliver.

What began as an assay challenge has become a story of breakthrough data. With Mabtech’s tools, Jeremy is moving closer to a vaccine that could protect against one of the world’s deadliest bacterial infections.