Fig. 3

Exosome production from cultured PBMCs. a Normal donor (ND) or HAM/TSP (HAM) PBMCs were cultured in exosome free media and measured over 2–5 days for ³H-thymidine uptake in counts per minute (CPM). NDs were left unstimulated by IL-2 or CD3 (ND unstim). b Carboxyfluorescein succinimidyl ester (CFSE) signal by HAM/TSP PBMCs was measured after 5 days in culture. One representative graph is pictured. c ELISAs were performed over 5 days for CD81 from nanotrapped (NT80 + 82) exosomes from ND and HAM PBMCs. ND PBMCs were maintained with IL-2 to induce proliferation. d Exosomes were nanotrapped (NT80 + 82) after 5 days culture of HAM or ND PBMCs (n = 5), followed by ELISA for CD81. ND PBMCs were maintained with IL-2 to induce proliferation. e A CD81 ELISA was performed from nanotrapped (NT80 + 82) exosomes isolated from HAM 1 and HAM 17 patient samples over 5 days in culture. f Vesicles isolated from HAM PBMC (HAM 4, 19, 21, 25, and 30) short-term cultures by NT pulldown were measured for AchE activity. g ND PBMCs were sorted immediately after thawing (IL-2 only) or after activation for 1 day with anti-CD3 100 ng/mL and 100 IU/mL IL-2 (activated). h HAM/TSP PBMCs were cultured for 24 h prior to FACS sorting into CD4⁺CD25⁻, CD4⁺CD25⁺, CD8⁺CD25⁻, and CD8⁺CD25⁻ T cells (n = 4). Statistical analysis was performed by unpaired t-test. *p-value < 0.05; **p-value < 0.01