Presenter: Ronald Germain, NIAID, NIH, USA
From the EMBO | EMBL Symposium: Seeing is Believing Imaging the Processes of Life
EMBL Advanced Training Centre Heidelberg, Germany 4 - 7 October 2017
Immune responses involve cell cell interactions within lymphoid tissues, trafficking of activated cells to sites of effector function, and the migration of effector cells within peripheral tissues. To gain insight into the relationships among cell movement, tissue architecture, and immune function, we have used intravital multiphoton microscopy and novel multiplex immunohistochemical methods we have developed called Histo cytometry and Ce3D. Observations: Innate immune (neutrophil) responses have been dissected at the molecular level and the role of cell death as a primary organizing factor in neutrophil swarming and secondary tissue damage uncovered, along with evidence of a macrophage dependent process of ‚Cloaking' that may protect tissues from cell injury under normal conditions. The role of cell localization in both innate and adaptive immunity has also been addressed using Histo cytometry in combination with a new clarification method called Ce3D. With an ability to use as many as 14 different colors and antibodies not only to surface markers but to phospho proteins and cytokines, and also to conduct imaging in large 3D volumes in a quantitative manner, our multiplex imaging technology facilitates analysis of the phenotype, number, location, signaling state, and function of immune cells and stromal elements in infected, inflamed, or tumor sites. Conclusion: This talk will illustrate the power of in situ imaging for the acquisition of a more accurate picture of the molecular, cellular, spatial, and temporal aspects of cell function and signaling events in host immune responses. This work was supported in part by the Intramural Research Program of the NIH, NIAID.
