The novel chelator lipid 3(nitrilotriacetic acid)-ditetradecylamine (NTA(3)-DTDA) promotes stable binding of His-tagged proteins to liposomal membranes: potent anti-tumor responses induced by simultaneously targeting antigen, cytokine and costimulatory signals to T cells.
Recent studies indicate that the chelator lipid nitrilotriacetic acid ditetradecylamine (NTA-DTDA) can be used to engraft T cell costimulatory molecules onto tumor cell membranes, potentially circumventing the need for genetic manipulation of the cells for development of cell- or membrane-based tumor vaccines. Here, we show that a related lipid 3(nitrilotriacetic acid)-ditetradecylamine (NTA(3)-DTDA, which has three NTA moieties in its headgroup instead of one) is several-fold more effective than NTA-DTDA at promoting stable His-tagged protein engraftment. IAsys biosensor studies show that binding of His-tagged B7.1 (B7.1-6H) to NTA(3)-DTDA-containing membranes, exhibit a faster on-rate and a slower off-rate, compared to membranes containing NTA-DTDA. Also, NTA(3)-DTDA-containing liposomes and plasma membrane vesicles (PMV) engrafted with B7.1-6H and CD40-6H exhibit greater binding to T cells, in vitro and in vivo. Engrafted NTA(3)-DTDA-containing PMV encapsulated cytokines such as IL-2, IL-12, GM-CSF and IFN-gamma, allowing targeted delivery of both antigen and cytokine to T cells, and stimulation of antigen-specific T cell proliferation and cytotoxicity. Importantly, use of B7.1-CD40-engrafted PMV containing IL-2 and IL-12 as a vaccine in DBA/2J mice induced protection against challenge with syngeneic tumor cells (P815 mammary mastocytoma), and regression of established tumors. The results show that stable protein engraftment onto liposomal membranes using NTA(3)-DTDA can be used to simultaneously target associated antigen, costimulatory molecules and cytokines to T cells in vivo, inducing strong anti-tumor responses and immunotherapeutic effect.
van Broekhoven
Christina L
CL
School of Biochemistry and Molecular Biology, Faculty of Science, The Australian National University, Canberra, ACT 0200, Australia.
Altin
Joseph G
JG
D000588:Amines / Q000737:chemistry*; D000818:Animals; D000941:Antigens / Q000378:metabolism; D018122:B7-1 Antigen / Q000737:chemistry; D015374:Biosensing Techniques; D019013:CD40 Antigens / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002614:Chelating Agents / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D006639:Histidine / Q000737:chemistry; D018664:Interleukin-12 / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D007700:Kinetics; D008055:Lipids; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D008956:Models, Chemical; D009368:Neoplasm Transplantation; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors; D014162:Transfection
D000588:Amines; D000941:Antigens; D018122:B7-1 Antigen; D019013:CD40 Antigens; D002614:Chelating Agents; D016207:Cytokines; D007376:Interleukin-2; D008055:Lipids; D008081:Liposomes; D011994:Recombinant Proteins; C405382:nitrilotriacetic acid ditetradecylamine; D018664:Interleukin-12; D006639:Histidine; D009571:Nitrilotriacetic Acid