The selective delivery of bioactive molecules (lipids, proteins and nucleic acid) during cognate T cell:APC interactions can occur via the exchange of specific nano-meter sized vesicles termed exosomes, during immunological synapse formation. To date, biochemical composition of exosomes, in particular those of primary cells involved within the immune response is largely unknown, although this information is fundamental to understanding exosomal biogenesis, release and cellular uptake. We recently showed that CD8+ T cells absent of Caveolin (Cav)1, a transmembrane protein involved in the regulation of membrane composition and trafficking, had significantly reduced cholesterol and sphingomyelin content. We observed in CD8+ T cells, Cav1 redistributed to lipid-cholesterol insoluble cellular fractions upon TCR-dependent activation and surprisingly was undetectable in in vitro CTLs. Isolation of exosomes from T cell culture media revealed enrichment of Cav1, in addition to other exosome-associated proteins Alix and TSG101. Cav1+ exosomes were also highly enriched for the Src-family kinases Lck and Fyn, and expressed canonical T cell markers; TCR, CD8 and CD25. Loss of Cav1 did not impact upon exosome morphology, surface charge, polydispersity or rigidity, although Cav1KO exosomes had a significant reduction in cholesterol content. Upon addition of exosomes to resting autologous CD8+ T cells, loss of Cav1 impacted upon exosomal fusion rate and intracellular uptake. These results indicate that Cav1 enrichment in CD8+ T cell exosomes contributes to cholesterol homeostasis and optimal exosome uptake.