Plasmacytoid dendritic cell line used in active or adoptive cell therapy
The present invention relates to a method for inducing and amplifying specific effectors, which comprises obtaining pulsed plasmacytoid dendritic cells (pDC) by incubation of a pDC line with at least one antigen, the pulsed pDC being subsequently irradiated and brought into contact with peripheral blood mononuclear cells (PBMC), and cultured or injected into an organism. The pulsed and irradiated pDC and the PBMC share at least one major histocompatibility complex (MHC) allele.
1. A method for inducing and amplifying antigen specific effectors in vitro, which comprises the following steps:
(a) obtaining pulsed plasmacytoid dendritic cells (pDCs) by incubation of a pDC line with at least one antigen,
(b) irradiating the cells obtained in (a),
(c) obtaining purified-peripheral blood mononuclear cells (PBMCs) from an individual, said PBMCs sharing at least one major histocompatibility complex (MHC) allele with the pulsed and irradiated pDCs obtained in step (b), and
(d) bringing the pulsed and irradiated pDCs obtained in step (b) into contact with the PBMCs obtained in step (c) and culturing said pulsed and irradiated pDCs with said PBMCs; and thereby inducing and amplifying antigen specific effectors in the culture in vitro, wherein the antigen specific effectors are CD8+ T lymphocytes.
2. The method for inducing and amplifying antigen specific effectors of claim 1 , which comprises at least one restimulation step consisting of
(e) bringing the cells obtained in step (d) into contact again with the pulsed and irradiated pDC line obtained in step (b), and culturing.
3. A method for inducing and amplifying antigen specific effectors in an individual in need thereof, which comprises the following steps:
(a) obtaining pulsed pDCs cells sharing at least one MHC allele with the PBMCs of said individual by incubation of a pDC line with at least one antigen,
(b) irradiating the cells obtained in step (a),
(c) injecting the pulsed and irradiated pDC cells obtained in step (b) into said individual,
the pDCs cells sharing at least one MHC allele with the PBMCs of said individual into which they are injected, and thereby inducing and amplifying antigen specific effectors in said individual, wherein the antigen specific effectors are CD8+ T lymphocytes.
4. The method for inducing and amplifying antigen specific effectors as claimed in claim 3 , in which the injection of the pulsed, irradiated pDCs of step (b) is repeated at least once.
5. The method for inducing and amplifying antigen specific effectors as claimed in claim 1 , wherein the at least one antigen is a tumor antigen, a microbial antigen, or a viral antigen.
6. The method for inducing and amplifying antigen specific effectors as claimed in claim 5 , wherein the antigen is a peptide.
7. The method for inducing and amplifying antigen specific effectors as claimed in claim 6 , wherein the peptide is chosen from the peptides included in the sequence of the tumor antigens CEA, NY-BR1, Her-2/Neu, PSA, RAGE-1, PRAME, TRP-2, MAGE-A1,MAGE-A2, MAGE-A4, MAGE-A9, MAGE-A10, MAGE-C2, MUC-1, p53, hTERT, survivin, melan-A/MART-1, GP100, tyrosinase, MAGE-A3 or NY-ESO1.
8. The method for inducing and amplifying antigen specific effectors as claimed in claim 6 , wherein the peptide is chosen from the peptides included in the sequence of the viral antigens env, nef, gp41, gp120, gag or pol of the HIV virus, HBc or HBs of the HBV virus, core, NS3 or NS5 of the HCV virus, Flu M1 of the influenza virus, pp65 of the CMV virus, BMLF1, LMP2, EBNA-2 or EBNA-3a of the EBV virus.
9. The method for inducing and amplifying antigen specific effectors as claimed in claim 1 , wherein the pDC line is obtained from pDC leukemia cells.
10. The method for inducing and amplifying antigen specific effectors as claimed in claim 9 , wherein the pDC line is the GEN2.2 line or the GEN3 line.
11. A method for treating cancers and/or infectious diseases, comprising
(a) obtaining pulsed plasmacytoid dendritic cells (pDCs) by incubation of a pDC line with at least one antigen,
(b) irradiating the cells obtained in (a),
(c) obtaining purified-peripheral blood mononuclear cells (PBMCs) from an individual in need of such treatment, wherein the PBMCs share at least one major histocompatibility complex (MHC) allele with the pulsed and irradiated pDCs obtained in step (b), and
(d) bringing the pulsed and irradiated pDCs obtained in step (b) into contact with the PBMCs obtained in step (c) and culturing said pulsed and irradiated pDCs with said PBMCs; and thereby inducing and amplifying antigen specific effectors in the culture in vitro, wherein the antigen specific effectors are CD8+ T lymphocytes; and
(e) injecting the antigen specific effectors into the individual in need of such treatment.
12. The method for inducing and amplifying antigen-specific effectors as claimed in claim 1 , which further comprises a step (e) consisting of:
(e) collecting the antigen-specific effectors induced and amplified in step (d).
13. The method of claim 11 , wherein the at least one antigen a tumor antigen, a microbial antigen, or a viral antigen.
14. The method of claim 11 , wherein the at least one antigen is a peptide included in the sequence of the antigens CEA, NY-BR1, Her-2/Neu, PSA, RAGE-1, PRAME, TRP-2, MAGE-A1, MAGE-A2, MAGE-A4, MAGE-A9, MAGE-A10, MAGE-C2, MUC-1, p53, hTERT, survivin, melan-A/MART-1, GP100, tyrosinase, MAGE-A3 or NY-ESO1.
15. The method of claim 11 , wherein the at least one antigen is a peptide included in the sequence of the antigens env, nef, gp41, gp120, gag or pol of the HIV virus, HBc or HBs of the HBV virus, core, NS3 or NS5 of the HCV virus, Flu M1 of the influenza virus, pp65 of the CMV virus, BMLF1, LMP2, EBNA-2 or EBNA-3a of the EBV virus.
16. The method for inducing and amplifying antigen specific effectors as claimed in claim 3 , wherein the at least one antigen is a tumor antigen, a microbial antigen, or a viral antigen.
17. The method for inducing and amplifying antigen specific effectors as claimed in claim 16 , wherein the antigen is a peptide.
18. The method for inducing and amplifying antigen specific effectors as claimed in claim 17 , wherein the peptide is chosen from a peptide included in the sequence of tumor antigens CEA, NY-BR1, Her-2/Neu, PSA, RAGE-1, PRAME, TRP-2, MAGE-A1, MAGE-A2, MAGE-A4, MAGE-A9, MAGE-A10, MAGE-C2, MUC-1, p53, hTERT, survivin, melan-A/MART-1, GP100, tyrosinase, MAGE-A3or NY-ESO1.
19. The method for inducing and amplifying antigen specific effectors as claimed in claim 17 , wherein the peptide is chosen from a peptide included in the sequence of viral antigens env, nef, gp41, gp120, gag or pol of the HIV virus, HBc or HBs of the HBV virus, core, NS3 or NS5 of the HCV virus, Flu M1 of the influenza virus, pp65 of the CMV virus, BMLF1, LMP2, EBNA-2 or EBNA-3a of the EBV virus.
20. The method for inducing and amplifying antigen specific effectors as claimed in claim 3 , wherein the pDC line is obtained from pDC leukemia cells.
21. The method for inducing and amplifying antigen specific effectors as claimed in claim 20 , wherein the pDC line is the GEN2.2 line or the GEN3 line.