Coformulation with tattoo ink for immunological assessment of vaccine immunogenicity in the draining lymph node

Barber-Axthelm, Isaac M; Kelly, Hannah G; Esterbauer, Robyn; Wragg, Kathleen; Gibbon, Anne; Lee, Wen Shi; Wheatley, Adam K; Kent, Stephen J; Tan, Hyon-Xhi; Juno, Jennifer A

Characterisation of germinal centre B and T cell responses yields critical insights into vaccine immunogenicity. Non-human primates are a key pre-clinical animal model for human vaccine development, allowing both lymph node and circulating immune responses to be longitudinally sampled for correlates of vaccine efficacy. However, patterns of vaccine antigen drainage via the lymphatics after intramuscular immunisation can be stochastic, driving uneven deposition between lymphoid sites, and between individual lymph nodes within larger clusters. In order to improve the accurate isolation of antigen-exposed lymph nodes during biopsies and necropsies, we developed and validated a method for co-formulating candidate vaccines with tattoo ink, which allows for direct visual identification of vaccine-draining lymph nodes and evaluation of relevant antigen-specific B and T cell responses by flow cytometry. This approach improves the assessment of vaccine-induced immunity in highly relevant non-human primate models.