Engineering immunomodulatory capacity on lymph node-targeted drug delivery carriers

• Dylan Ngo, Human Physiology, University of Delaware

• Jason Gleghorn, Biomedical Engineering, University of Delaware

Dylan T. Ngo¹, Omkar S. Mhatre², Michael J. Donzanti², Jason P. Gleghorn²

College of Arts and Sciences¹ and Department of Biomedical Engineering², University of Delaware, Newark, DE 19717

Lymph nodes (LNs) are vital immunological organs that feature complex structures and functions. Transport into the LN is extremely selective, which means that small molecules are largely excluded from the lobules. This makes targeted delivery of oncology drugs and cancer vaccines into the LN challenging for treatment as well as prevention of further metastasis. Lymph node metastasis is an important part of disease progression as untreated cancer cells can condition LN resident lymphocytes to tolerate their presence, further priming distant tissues to become amenable to metastatic colonization. Thus, the ability to overcome the lymph node barrier to deliver an immunomodulatory agent to activate LN resident cells could help confer protective immunity in cancer. To enhance therapeutic penetration into the LN lobule, our lab has developed cell-mimetic vehicles called cryo-shocked T lymphocytes (CSTLs) that leverage a Trojan-horse strategy to deliver drugs to the lymph node. As an extension of our current platform, we investigated CSTL’s capacity as a local immunomodulatory agent in the LN. Through surface modifications and directed activation of CSTLs, our objective was to adapt the delivery vehicle itself for stimulation of target immune cells in the LN by inducing cell-cell interaction and maturation marker expression. Using a pro-inflammatory signaling molecule and flow cytometry analysis, we demonstrated the expression of a T cell surface maturation marker on primary splenocytes in vitro. This same stimulatory agent was used to pre-activate CSTLs. Pre-activated CSTLs induced enhanced cell-cell interactions and grouping with cultured T cells compared to untreated controls, leading to higher frequency, prolonged CSTL-cell contact. Finally, we demonstrated the capability of attaching functional molecules of interest to the CSTL surface using a liposome patching strategy. A vehicle that efficiently delivers payload and directs immune function would serve as a potent therapeutic agent in the lymph node.