Abstract

Multiplex Immunofluorescence for Evaluation of NK Cell Therapy in a Murine Model of Glioblastoma Multiforme

Submitted and written by Samantha Hicks, Texas A&M University

Hypothesis: We hypothesized that multiplex immunofluorescence could be utilized to evaluate NK cell infiltration in both tumor and adjacent brain tissue of immunodeficient NSG mice engrafted with glioblastoma multiforme (GBM) tumor cells. 

Significance: Presently, most adoptive cell transfer (ACT) therapies are inconsistently effective in solid tumors and are solely labeled for hematogenous neoplasia. One emerging ACT therapy utilizes NK cells, an immune cell that has a natural predilection for targeting and destroying tumor cells. NK cell therapy is showing significant promise for the treatment of solid tumors such as GBM, the most common primary brain tumor in human adults. There still exists a considerable need for improved immunotherapies for solid tumors and the development of novel ACT therapies with improved efficacy and toxicity profiles.

Methods: We evaluated genetically engineered NK cell therapy for persistence and expansion in the tumor microenvironment and its effects on normal brain and GBM tumor tissue. In this study, NSG mice were cerebrally implanted with GSC-20 patient-derived GBM cells transduced to express firefly luciferase and genetically engineered or non-transduced NK cells with one cohort given only tumor cells. Mice were monitored with in vivo bioluminescent imaging of tumor cells. Mice were then euthanized, and the engrafted tumors and brain tissue were collected, formalin-fixed, paraffin-embedded, and stained using H&E and Akoya Biosciences Opal 7-color kit for multiplex immunofluorescence (mIF). Antibodies for granzyme B, caspase 3, Ki-67, Iba-1, luciferase, and GFAP were utilized and evaluated. The mIF highlighted microglia, GBM cells, NK cells, astrocytes, apoptotic cells, and proliferating cells. Whole slides were scanned using an Aperio AT2 and Leica Versa 8. We used Halo v3.6.4 to quantify phenotypes of interest and determine spatial relationships of microglia and NK cells, which synergistically work to produce an inflammatory response in the brain. 

Conclusion: NK cells demonstrated extensive tumor infiltration and local brain inflammation. NK cell therapy shows strong promise for treating solid tumors and mIF may be a useful tool for evaluating NK cell persistence and proliferation in tumors and the tumor microenvironment.  

Cutest Pet

Submitted by Bei Li Martin, St. George's University

Externs on the Hill

Submitted by Cristina Blanco, University of Minnesota

Meeting with North Carolina Representative Don Davis to discuss agriculture.

As the American Veterinary Medical Association Government Relations Extern for Spring 2024, I have had the amazing opportunity to watch veterinary medicine and policy merge to work toward the betterment of animals and humans throughout our country.

So far, I have been able to meet many veterinarians who work on Capitol Hill and within a variety of federal agencies including the U.S. Department of Agriculture (USDA), U.S. Agency for International Development (USAID) and the U.S. Fish and Wildlife Service (USFWS). Many also had experience as fellows for the American Association for the Advancement of Science (AAAS). Each of these veterinarians had a unique perspective on how their medical training relates to important policy work. For example, I met with veterinarians at USAID who  utilize their training toward a multisystem approach to global health. Their veterinary background is incredibly important in addressing global food shortages. Each day, more than 800 million people across the world lack enough food to eat. Their work with Feed the Future Innovation Labs joins USAID with universities to address global agriculture and food security issues. By understanding the details and importance of food animal production systems, veterinarians play a pivotal role in providing the global community with a safe protein source. 

I also had the wonderful opportunity to watch the floor proceedings of the House of Representatives.  These proceedings included discussions involving a wide range of bills from national security to wildlife conservation.  Each representative spoke for their constituents, supporting or disagreeing with bills. Some bills, like the Explore Act that expands outdoor recreation opportunities, were supported by representatives from across the United States.  

With the policy experts at the AVMA, I was able to meet with congressional offices to discuss bills that are important to the veterinary profession. These include the Rural Veterinary Workforce Act (H.R. 4355/ S.2829), which allows for the recruitment and retention of rural veterinarians, and the Healthy Dog Importation Act (H.R. 1184/ S. 502), which strengthens guidelines for the safe importation of dogs to prevent infectious disease. I was also able to help advocate for federal funding that would increase veterinary services in underserved areas by aiding veterinarians with educational loan debt through the Veterinary Medicine Loan Repayment Program and Veterinary Services Grant Program. The AVMA is also working to secure additional funding for the National Wildlife Health Center in Wisconsin. 

By discussing these issues with congressional staff, we were able to communicate the veterinary perspective on public policy that directly impacts the profession. It has truly been an amazing experience to see how policy and veterinary medicine merge in the world of advocacy.  

Scholarship Opportunity

Applications Open for the 2024 WSAVA Michael J. Day Scholarship  

Veterinary students with an interest in companion animal infectious disease, clinical vaccinology or immunology encouraged to apply  

The Scholarship provides financial assistance towards a research project into an aspect of small companion animal infectious disease, clinical vaccinology or immunology. All veterinary students studying towards a registerable veterinary degree in an accredited veterinary establishment in a country in which the WSAVA has a member association are eligible to apply for the Scholarship. Full details and guidance on the application process, together with eligibility criteria are available here: https://wsava.org/resources/michael-j-day-scholarship/

The deadline for applications for the 2024 Michael J. Day Scholarship is July 31, 2024. The successful Scholar will be informed by the end of October 2024. Their research project is expected to be conducted between January and September 2025. Within three months of completion, the Scholar will be expected to provide a 1,500-word report on their findings. They will also be invited to present or publish their findings in English, as a poster or abstract, during WSAVA World Congress.

Student Experience

After getting into vet school they brought my experience to a whole new level as the doctors there begin to mentor me in beginning surgeries with shelter dogs. They have taught me how to perform these surgeries and given me the opportunity to further my knowledge inside and out. I have performed multiple surgeries now under their watch and the passion I have for vet med has only bloomed more for me. I guess in the beginning this was considered a job and turned into more of an externship. But regardless for me it has been the opportunity of a lifetime because it’s helped shape me into the person I am today. Someone that steps out of their comfort zone and into the light with a scalpel blade. Daunting, yes but I couldn’t have asked for a better experience that has only made me drive more towards the dream that I’ve always wanted. Not only that but along the way I have gained a second family that has been so supportive of me. I’ve learned to work under circumstances that are unimaginable. Some of these experiences I didn’t realize how important they were to me until this opportunity showed me that I am meant to be a veterinarian because that’s where my passion lies with no doubt. This has been a very meaningful part of my veterinary experience and one that I hope to continue in the future.

Click to read more ...