Protocol for the characterization of autoproliferating B and T cells from persons with multiple sclerosis

Autoproliferation (AP) is the spontaneous in vitro activation and proliferation of B- and T cells in the absence of exogenous stimulation. Here, we present a protocol to examine AP using peripheral blood mononuclear cells (PBMCs) derived from persons with multiple sclerosis (pwMS). We describe steps for the isolation of autoproliferating (AP+) lymphocytes as well as the expansion of AP⁺ T cells for downstream analyses. We detail procedures for standard fluorescence-activated cell sorting (FACS) and cell culturing. For complete details on the use and execution of this protocol, please refer to Jelcic et al.^1,2.