ENHANCING THE T-CELL STIMULATORY CAPACITY OF HUMAN ANTIGEN PRESENTING CELLS IN VITRO AND IN VIVO AND THEIR USE IN VACCINATION
New methods of in vitro or in vivo enhancing the T-cell stimulatory capacity of human dendritic cells (DCs) and the use thereof in cancer vaccination are provided. The method includes introduction of different molecular adjuvants to human DCs by contacting or modifying them with mRNA or DNA molecule(s) encoding CD40L, and CD70 or constitutively active TLR4 (caTLR4).
1 . A method for inducing an antigen-specific immune response in a subject, comprising the step of administering to said subject:
a) one or more mRNA or DNA molecule(s) encoding the functional immunostimulatory proteins CD40L, CD70, and caTLR4, and
b) target-specific antigen(s).
2 . The method of claim 1 , wherein said mRNA or DNA molecules and target-specific antigens are administered to the lymph node(s), to the tumor, or wherein said mRNA or DNA molecules are administered subcutane, intradermally or intravenously.
3 . The method of claim 2 , wherein said mRNA or DNA molecules and target-specific antigens are administered through intranodal, intratumoral, subcutane or intradermal injection or through intravenous administration.
4 . The method of claim 1 , wherein the target-specific antigen is a tumor antigen.
5 . The method of claim 1 , wherein the target-specific antigen is a bacterial, viral or fungal antigen.
6 . The method of claim 1 , wherein said target-specific antigen is selected from the group consisting of: total mRNA isolated from (a) target cell(s), one or more specific target mRNA molecules, protein lysates of (a) target cell(s), specific proteins from (a) target cell(s), a synthetic target-specific peptide or protein, and synthetic mRNA or DNA encoding a target-specific antigen or its derived peptide(s).
7 . The method of claim 1 , wherein said subject is suffering from a disease or disorder selected from the group consisting of: neoplastic disorders, infectious disorders, or immunological disorders.
8 . The method of claim 7 , wherein said subject is suffering from a disease or disorder selected from the group consisting of: tumor presence, cancer, melanoma presence, bacterial infection, viral infection, fungal infection, HIV infection, hepatitis infection, immunosuppression, SCID, and AIDS.
9 . The method of claim 1 , additionally comprising the administration of mRNA or DNA encoding one or more of the following molecules: IL-12p70, EL-selectin, CCR7, and/or 4-1BBL.
10 . The method of claim 1 , additionally comprising the administration of molecules inhibiting SOCS, A20, PD-L1 or STAT3 expression or function.
11 . A vaccine comprising:
a) one or more mRNA or DNA molecule(s) encoding the functional immunostimulatory proteins CD40L, CD70, and caTLR4, and
b) target-specific antigen.
12 . The vaccine of claim 11 , wherein said target-specific antigen is selected from the group consisting of: total mRNA isolated from (a) target cell(s), one or more specific target mRNA molecules, protein lysates of (a) target cell(s), specific proteins from (a) target cell(s), a synthetic target-specific peptide or protein, and synthetic mRNA or DNA encoding a target-specific antigen or its derived peptide(s).
13 . The vaccine of claim 11 , wherein said target-specific antigen is a tumor antigen.
14 . The vaccine of claim 11 , wherein the target-specific antigen is a bacterial, viral or fungal antigen.
15 . The vaccine of claim 11 , wherein said mRNA or DNA molecules encoding the immunostimulatory proteins are part of a single mRNA or DNA molecule.
16 . The vaccine of claim 15 , wherein the single mRNA or DNA molecule is capable of expressing the two or more proteins simultaneously.
17 . The vaccine of claim 16 , wherein the mRNA or DNA molecules encoding the immunostimulatory proteins are separated in the single mRNA or DNA molecule by an internal ribosomal entry site (IRES) or a self-cleaving 2a peptide encoding sequence.
18 . The vaccine of claim 11 , additionally comprising mRNA or DNA molecules encoding one or more of the proteins selected from the group consisting of: IL-12p70, EL-selectin, CCR7, and 4-1BBL.
19 . The vaccine of claim 11 , additionally comprising one or more molecules inhibiting SOCS, A20, PD-L1 or STAT3 expression or function.
20 . The vaccine method according to claim 11 , wherein said mRNA is protected.
21 . The method according to claim 1 , wherein said mRNA is protected and administered intravenously.