Demyelinated and inflammatory lesions in the multiple sclerosis (MS) brain contain immune cells with variable phenotypes and functions. Standard approaches restrict the number of analytes one can examine in a single tissue section making it difficult to identify complex cell phenotypes and, simultaneously, their relation to tissue injury. 

Here, we used imaging mass cytometry (IMC) to enable the simultaneous imaging of 15+ proteins within staged MS lesions. To test the potential for IMC to discriminate between different types of lesions, we selected a case with severe rebound MS disease activity after natalizumab cessation.

With post-acquisition analysis pipelines we were able to: (1) Discriminate demyelinating macrophages from the resident microglial pool; (2) Determine which types of lymphocytes reside closest to blood vessels; (3) Identify multiple subsets of T and B cells, and (4) Ascertain dynamics of T cell phenotypes vis-à-vis lesion type and location.

We are currently using IMC to obtain a comprehensive analysis of single-cell phenotypes in relation to lesion morphometry in well-characterized brain tissue from a large cohort of MS patients.