A membrane penetrating multiple antigen peptide (MAP) incorporating ovalbumin CD8 epitope induces potent immune responses in mice

作者：Brooks Nicole A; Pouniotis Dodie S; Sheng Kuo Ching; Apostolopoulos Vasso; Pietersz Geoffrey A^*

来源：Biochimica et Biophysica Acta-Biomembranes, 2010, 1798(12): 2286-2295.

DOI：10.1016/j.bbamem.2010.05.007

• Cell penetrating peptides
• Penetratin
• Cancer vaccines
• Multiple antigen peptide
• Antigen presentation

Cell penetrating peptides (CPP) represent a novel approach to facilitate cytoplasmic delivery of macromolecules. The DNA binding domain of Drosophila Antennapedia contains 60 amino acids and consists of 3 alpha-helices, with internalizing activity mapped to a 16-amino acid peptide penetratin (Antp) within the third alpha-helix. Here, we report on the use of penetratin to deliver a multiple antigen peptide (MAP) incorporating the immunodominant COB epitope of ovalbumin, SIINFEKL (MAPOVACD8). We demonstrate that penetratin linked to the MAPOVACD8 construct either by a disulfide (SS) or thioether (SC) linkage promotes the uptake, cross presentation and subsequent in vivo proliferation and generation of OVACD8 (SIINFEKL)-specific T cells. The MAPOVACD8 construct without penetratin is not presented by MHC class I molecules nor does it generate an in vivo IFN-gamma response in C57BL/6 mice. Moreover, we clearly define the uptake and intracellular processing pathways of AntpMAPOVACD8 SS and SC revealing the majority of AntpMAPOVACD8 is taken up by DC via an endocytic, proteasome and tapasin independent mechanism. We also show that the uptake mechanism of AntpMAPOVACD8 is dose dependent and uptake or intracellular processing is not altered by the type of chemical linkage.

• 出版日期2010-12