Evaluating Innate Immune Response in Allergic Patients Using Temperature Controlled Basophil Activation Testing

Overview

Objectives/Goals This study aimed to understand the effect of temperature on basophilic activation, as well as the effect of environmental factors during an allergic reaction. Methods/Materials In order to assess the severity of an allergic reaction, the developed Basophil Activation Test Assay (BAT Assay) was implemented. Whole blood taken from peanut-allergic patients was incubated with RPMI (negative control, growth hormone), anti-IgE (positive control), and peanut extract. After incubation, degranulation is stopped and cells are stained with fluorescent antibodies. On basophils, CD63 and CD203c are surface proteins which only become expressed during activation/degranulation. Thus by using CD203c-PE and CD63-APC, we tag the activated basophils which can later be recognized and isolated by flow cytometry. Using the flow cytometer, the cell poplulation from the whole blood sample is narrowed down to only basophils. In this isolated population, the percentage of activated basophils indicates the severity of the reaction. By completing multiple BAT Assays at room temperature, 37, 38.6, and 42 degrees Celsius, the trend of reactivity over temperature can be accurately assessed. Results Looking at the averages of the compiled data, the negative control as well as the positive control at higher temperatures resulted in less activation. Meanwhile, the peanut extract prompted a stronger allergic response at higher temperatures. The samples completed at room temperature and 42˚C varied too much to determine the trends. Conclusions/Discussion Because the peanut extract samples were more reactive at higher temperatures, unlike the anti-IgE samples, this suggests that peanut allergies may be non-IgE mediated at these higher temperatures. These results are imply that allergic pathways may be triggered differently due to blood temperature. Through this, more effective methods of treatment can target certain pathways at certain temperatures. The hypothesis was partially supported; peanut extract was indeed more reactive at a higher temperature, whereas the anti-IgE and RPMI samples were not.

Awards (1)

• Honorable Mention

Competition history

• CSEF 2015 Mammalian Biology · Entry S1221

Resources

• CSEF project summary ↗

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