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    of immune responses (Balasse et al., 2008). Balasse et al. reported that the injection route of nanoparticles affected the quality of immune re-sponses due to differences in frequency and population type of APCs at the injection sites. During intraperitoneal (i.p) and s.c immunization, the injected particles are mainly taken up by APCs and DCs of perito-neal cavity and subcutaneous, while in i.v administration minimum amount of the particles are immediately available for APC and DCs (Balasse et al., 2008). Accordingly, in our present study, F4 liposomal formulation which was administrated through intravenous route in-duced lower level of IFN-γ compared to F2- to F3-liposomes that ad-ministrated through subcutaneous route.
    Nonetheless, the survival analysis study showed neither cancer immunotherapy nor significant side effect resulting in death event. The in-vivo results revealed that F1-liposome induces a phospholipid-spe-cific cellular immune response, although the generated immune re-sponse against phospholipid content of liposome could not induce any targeted immunity against implanted tumor which leads to enhanced survival.
    5. Conclusion
    In the present study, four different liposomal formulation platforms were investigated in terms of cytotoxicity, immunogenicity and ther-apeutic activity. Among the liposomal formulations, the liposomal formulations containing DOTAP as a positively charged phospholipid and DOPE were able to stimulate cellular immunity and CTLs responses such as IFN-γ secretion as well as stimulation of IL-4 and IL-17. Taken together, fluid DOTAP and DOPE-containing liposomes can stimulate a mixture of Th1 and Th2-immune responses in an antigen-free manner.
    The technical supports of Nanotechnology research center, Mashhad University of Medical Sciences are acknowledged.
    The financial supports of Mashhad University of Medical Sciences are acknowledged.
    Declarations of Competing Interest
    The authors declare that no competing interest, financial or other-wise exists in relation to the present study.
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