Characterization of hybrid CTL epitope delivery systems consisting of the Antennapedia homeodomain peptide vector formulated in liposomes.
Peptide carriers, such the homeodomain of Antennapedia molecule (AntpHD), which spontaneously cross cellular membranes, have been exploited to deliver antigenic peptide Cw3 to the major histocompatibility complex (MHC) class-I presentation pathway and to prime cytotoxic T cells (CTL). However, the in vivo use of AntpHD recombinant peptide has been limited because CTLs can only be primed in the presence of sodium dodecyl sulfate (SDS) as adjuvant. In this report, we have exploited liposomes to protect the AntpHD-Cw3 from serum degradation and to facilitate the delivery of the recombinant peptide into the MHC class-I pathway of antigen-presenting cells. We have demonstrated that AntpHD recombinant peptide spontaneously associates with liposomes and this association is stable in vitro. However, exchange studies assessing the transfer of the peptide to model membranes or cells in vitro indicates that approximately 50% of the liposome-associated peptide is readily exchangeable. This is consistent with trypsin-protection assays, which have shown that approximately 40% of the liposome-associated peptide is protected from hydrolysis. Importantly, macrophages and dendritic cells are able to internalize AntpHD recombinant peptide associated with liposomes resulting in efficient delivery of the CTL peptide into the cytosol. These studies have demonstrated that dendritic cells treated with AntpHD-Cw3 in liposomes sensitize CTL clones to lyse syngeneic target cells expressing Cw3 epitope. This strategy, which combines liposomes and a peptide vector, provides a new approach for introducing molecules into the MHC class-I antigen presentation pathway of dendritic cells.
Chikh
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Systemic Therapy Program, Advanced Therapeutics, British Columbia Cancer Agency, 600 West 10th Avenue, Vancouver, BC, Canada V5Z 1L3.
Bally
M
M
Schutze-Redelmeier
M P
MP
D000818:Animals; D051943:Antennapedia Homeodomain Protein; D017951:Antigen Presentation / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000276:immunology*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D016503:Drug Delivery Systems; D004705:Endocytosis / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005822:Genetic Vectors; D015236:HLA-C Antigens / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D018398:Homeodomain Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D008081:Liposomes; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins; D010455:Peptides / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014157:Transcription Factors
D051943:Antennapedia Homeodomain Protein; D018984:Epitopes, T-Lymphocyte; D015236:HLA-C Antigens; C038653:HLA-C*03 antigen; D018398:Homeodomain Proteins; D008081:Liposomes; D009687:Nuclear Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors