Caty Tylan measuring anole footpad thickness as a indicator of immune function.
A major challenge for organisms is to protect themselves from pathogens, things in the environment, including other organisms or toxins, that can cause disease and harm them. Animals, including anoles, have several different types of immune responses by which they can respond to pathogens. Ecologists are particularly interested in how these immune responses work in natural settings, how effective they are at protecting organisms, and how other aspects of an organism’s environment, including diet, stress, and reproduction, may positively or negatively impact immunity. However, immune systems are very complex, and measuring immune function, especially in the field, can be quite challenging!
Ecoimmunologists have developed various procedures to test different aspects of immune function, but ideally these procedures should be validated, or proved to be meaningful, in each organism they are used in. The phytohemagglutinin (PHA) skin test is one such assessment of immune function that is commonly used. This procedure involves injecting a small amount of phytohemagglutinin, a plant protein that provokes an immune response in animals, into the tissue of an organism of interest, waiting a given amount of time, and measuring the resulting swelling which is then used as an index of immune response. Researchers like this test because it is minimally invasive, works in almost any species, and is simple and easy to do in the field. Though this test has been used in multiple reptile species, it has never been validated in a reptile.
Enter Caty Tylan, a Ph.D. student and diploma-carry veterinarian from Penn State University, who addressed this issue in her talk “Local and systemic immune response to phytohemagglutinin: Validation of the PHA skin test in the green anole, Anolis carolinensis.” Caty chose to validate this test in green anoles (Anolis carolinensis), a model anole species, using two different types of PHA, PHA-L and PHA-P, which are available to researchers. She injected PHA into anole footpads and compared swelling in those feet over 48 hours post-injection. By comparing swelling in these feet to those which were only injected with sterile saline, Caty showed that PHA does induce a swelling response over 48 hrs, and that this response is the same for both types of PHA. She also examined the white blood cell counts in these anoles and found that PHA-L, a more purified and specific PHA, induced the stronger lymphocyte response, an immune measure that many ecoimmunologists look to quantify. In the future, Caty will examine histological sections of injected anole feet to examine the local immune response to PHA injections and fully validate this assay. This work should allow effective use of the PHA assay in future anole research and support investigations into how various environmental variables affect cell-mediated immune function in reptiles.
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