Phototruncation cell tracking with near-infrared photoimmunotherapy using heptamethine cyanine dye to visualise migratory dynamics of immune cells

Background Noninvasive in vivo cell tracking is valuable in understanding the mechanisms that enhance anti -cancer immunity. We have recently developed a new method called phototruncation-assisted cell tracking (PACT), that uses photoconvertible cell tracking technology to detect in vivo cell migration. This method has the advantages of not requiring genetic engineering of cells and employing tissue -penetrant near -infrared light. Methods We applied PACT to monitor the migration of immune cells between a tumour and its tumour -draining lymph node (TDLN) after near -infrared photoimmunotherapy (NIR-PIT). Findings PACT showed a significant increase in the migration of dendritic cells (DCs) and macrophages from the tumour to the TDLN immediately after NIR-PIT. This migration by NIR-PIT was abrogated by inhibiting the sphingosine-1-phosphate pathway or G alpha i signaling. These results were corroborated by intranodal immune cell profiles at two days post -treatment; NIR-PIT significantly induced DC maturation and increased and activated the CD8+ T cell population in the TDLN. Furthermore, PACT revealed that NIR-PIT significantly enhanced the migration of CD8+ T cells from the TDLN to the tumour four days post -treatment, which was consistent with the immunohistochemical assessment of tumour -infiltrating lymphocytes and tumour regression. Interpretation Immune cells dramatically migrated between the tumour and TDLN following NIR-PIT, indicating its potential as an immune -stimulating therapy. Also, PACT is potentially applicable to a wide range of immunological research.