Table 6.

Functional role for EVs in drug-induced liver injury.

Model/EV source	Dose	Isolation method	EV marker	Result	References [#]
RPMI 8866 B lymphocytes Cell media	0.5 mg/mL Biotinylated amoxicillin/amoxicillin	200,000 g UC	None reported	AX-EVs contain haptenated HSP70, EF-2, actin, α-enolase. AX-EVs were internalized by B lymphocytes	[169]
Long Evans Plasma	0.8 mg/kg PAH via oral gavage 3/wk. for 90 days	100,000 g UC	ALIX/TSG101/CD63/CD81/Flotillin1	Chronic PAH treatment leads to increased EVs in plasma	[165]
PRH WIF-B9	100 nmol/L BP, DBA, or PYRa	100,000 g UC	ALIX/TSG101/CD63/CD81/Flotillin1	BP and DBA require AhR activation to increase EV release. PYR functions through CAR. BP and DBA increase plasma membrane fluidity. Increase in EV release is independent of apoptosis	[165]
HMEC-1 Cell media	B[a]P 1–10,000 nmol/L	10,000 g UC (large EVs)   100,000 g UC (sEVs)	TSG101, Caveolin-1, Flotillin-1, Hsc70   CD63, TSG101, Caveolin-1, Flotillin-1, Hsc70	AhR inhibitor (10 μmol/L) prevents B[a]P induced sEV release without effecting large EVs Suggests that EV subpopulations are differentially impacted by B[a]P	[167]
PHH	AX 0.05 mmol/L FX 0.05 mmol/L Isoniazid 0.03 mmol/L SMX-NO 0.01 mmol/L Collected 24 h later	Exoquick-TC (commercial)	CD63b	AX-, FX-, and SMX-NO-EVs contain haptenated proteins. Dendritic cells sequester drug-EVs via phagocytosis AX-EVs fail to activate dendritic cells	[170]

^aPRH were treated for 2–18 h, WIF-B9 hepatocytes were treated 5–72 h.

^bDrug-EVs were CD9, CD63, CD81, Hsp70 positive via mass spectrometry, western blot only detected CD63. HMEC1 is a human endothelial cell line. AX, amoxicillin, FX, flucloxacillin, SMX-NO, nitroso-sulfamethoxazole.