Although the lymphatic system plays an integral role in regulating immune responses throughout the body, it has the potential to compromise a patient’s health by providing tumor cells with the optimal environment for metastasis. As such, lymph node removal has been the standard of care procedure for head and neck squamous cell carcinoma (HNSCC) patients to prevent further cancer spread. However, with growing applications of immunotherapies towards enhancing a patient’s immune system to fight cancer, lymphoablation has rendered a population of HNSCC patients incapable of receiving the full potential of these novel therapies. Given that lymphoablation will remain as the standard practice for this type of cancer, a tissue engineered artificial lymph node may compensate for the opposing effects of the standard of care against immunotherapy treatment and improve treatment outcomes for patients.

As such, we have engineered a functional artificial lymph node to replace those lost in surgery, with the aim to revitalize the immune system’s activity and restore normal lymphatic vasculature. Our experimental results indicate that decellularizing a lymph node to industry standard of less than 50 ng dsDNA / mg ECM while retaining its protein scaffold is possible, but can be variable depending on the size of lymph nodes used. Furthermore, the resulting decellularized scaffold can successfully be seeded with primed dendritic cells that remain functional and capable of activating antigen-specific cytotoxic T lymphocytes (p<0.0001). IMARIS 3D imaging software was utilized to elucidate changes in lymphatic vasculature between mice with and without tumors, as well as those that underwent lymphatic surgery, which will be essential in analyzing the scaffold’s ability to reconnect to native vasculature after implantation in future in vivo studies.

We would like to thank Dr. Robert Saddawi-Konefka, Dr. Karen Christman, Catherine Watson, and Dr. Bruce Wheeler for providing their expertise and guidance to us throughout the course of this project. We would also like to thank the staff at the Moores Cancer Center, Sanford Consortium for Regenerative Medicine, and La Jolla Institute for Immunology, for the use of their facilities and resources for this project.