Topical vaccination with super-stable ready to use nanovesicles

- Ultradeformable nanovesicles containing archaeolipids (UDA) resist heat sterilization and storage under cold-free conditions.
- UDA do not require complex steps of antigen loading for eliciting immune response.
- Sterilized and lyophilized UDA mixed with antigen topically applied elicit systemic immune response.

Ultradeformable archaeosomes (UDA) are nanovesicles made of total polar archaeolipids (TPA) from the archaea Halorubrum tebenquichense, soybean phosphatidylcholine and sodium cholate (3:3:1 w/w). Fresh dispersions of UDA including different type of antigens are acknowledged as efficient topical vaccination agents. UDA dispersions however, if manufactured for pharmaceutical use, have to maintain colloidal stability upon liposomicidal processes such as sterilization and lyophilization (SLRUDA), needed to extend shelf life during storage. The remaining capacity of SLRUDA to act as adjuvants was therefore tested here for the first time. Another unexplored issue addressed here, is the outcome of replacing classical antigen inclusion into nanovesicles by their physical mixture. Our results showed that UDA behaved as super-stable nanovesicles because of its high endurance during heat sterilization and storage for 5 months at 40 °C. The archaeolipid content of UDA however, was insufficient to protect it against lyophilization, which demanded the addition of 2.5% v/v glycerol plus 0.07% w/v glucose. No significant differences were found between serum anti-ovalbumin (OVA) IgG titers induced by fresh or SLRUDA upon topical application of 4 weekly doses at 600 μg lipids/75 μg OVA to Balb/c mice. Finally, SLRUDA mixed with OVA elicited the same Th2 biased plus a non-specific cell mediated response than OVA encapsulated within UDA. Concluding, we showed that TPA is key component of super-stable nanovesicles that confers resistance to heat sterilization and to storage under cold-free conditions. The finding of SLRUDA as ready-to-use topical adjuvant would lead to simpler manufacture processing and cheaper products. .

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