Multilevel human secondary lymphoid immune system compartmentalization revealed by complementary imaging approaches

Secondary human lymphoid tissue immune reactions take place in a highly coordinated environment with compartmentalization representing a fundamental feature of this organization. In situ profiling methodologies are indispensable for the understanding of this compartmentalization. Here, we propose a complementary experimental approach aiming to reveal different aspects of this process. The analysis of human tonsils, using a combination of single cell phenotypic analysis based on flow cytometry and multiplex imaging and mass spectrometry based methodologies, revealed a compartmentalized organization at the cellular and molecular levels. More specifically, the skewed distribution of highly specialized immune cell subsets and relevant soluble mediators was accompanied by a compartmentalized localization of several lipids across different anatomical areas of the tonsillar tissue. The performance of such combinatorial experimental approaches could lead to the identification of novel in situ interactions and molecular targets for the in vivo manipulation of lymphoid organ, particularly the germinal center, immune reactions.

Automated summary

The secondary immune reactions in human lymphoid tissues require a highly coordinated and compartmentalized environment. Through a combination of flow cytometry, imaging, and mass spectrometry techniques, researchers have identified a compartmentalized organization at the cellular and molecular levels in human tonsils. This organization is characterized by the uneven distribution of specialized immune cell subsets, soluble mediators, and lipids across different anatomical areas of the tonsillar tissue. The application of these experimental approaches could lead to the discovery of novel in situ interactions and molecular targets for manipulating lymphoid organ immune reactions, particularly in the germinal center.