Allogeneic tumor therapeutic agent, a vaccine using allogeneic tumor cells for the therapeutic treatment of tumor diseases, and a method for the making of such a vaccine, and transfected human tumor cells for use as a vaccine
View Patent ↗A vaccine on the basis of allogeneic tumor cells for the therapeutic treatment of defined tumor diseases, and a method for the making of such vaccine is disclosed. The tumor cells were previously transfected ex vivo with expression constructs encoding cytokines and co-stimulatory factors.
1. A vaccine for the treatment of patients who suffer from tumor diseases, said vaccine containing transfected tumor cells of a genetically not identical (different) donor of the same species (allogeneic), wherein the tumor cells were transfected ex-vivo with nucleic acid molecules encoding:
interleukin-7 (IL-7), granulocyte-macrophage colony stimulating factor (GM-CSF), CD40L/CD154 and B7.1/CD80.
2. The vaccine according to claim 1 , wherein the encoding nucleic acid molecules are present in one or more expression constructs.
3. The vaccine according to claim 2 , wherein the expression construct is one of:
a) a plasmid; or
b) a linear double stranded, covalently closed expression cassette comprising essentially solely a CMV promoter, an intron, the encoding gene sequence and a polyadenylation sequence being closed on both ends of the double strand by a short loop of single stranded nucleotide residues.
4. The vaccine according to claim 3 , wherein:
said vaccine comprises additionally an immune modulating oligodeoxyribonucleotide as an adjuvant; and the immune modulating oligodeoxynucleotide:
a) comprises a sequence of the base sequence N 1 N 2 CGN 3 N 4 , where N 1 N 2 is an element selected from the group comprising GT, GG, GA, AT or AA, N 3 N 4 is an element selected from the group comprising CT or TT, and C is desoxycytosine, G is desoxyguanosine, A is desoxyadenosine and T is desoxythymidine,
b) and comprises a circular strand deoxyribonucleic acid with a partially complementary antiparallel base sequence and is of a dumbbell shape.
5. The vaccine according to claim 4 , wherein the sequence with the base sequence N 1 N 2 CGN 3 N 4 is positioned in the single stranded region of the oligodeoxyribonucleotide and comprises 40 to 200 nucleotides; and
said vaccine comprises a pharmaceutically acceptable carrier.
6. The vaccine according to claim 1 , wherein the allogeneic tumor cells are selected from colorectal carcinoma, small cell or non-small cell lung carcinoma, prostate carcinoma, mamma carcinoma, ovarian carcinoma, renal cell carcinoma and malign melanoma.
7. The vaccine according to claim 5 , wherein the tumor cells are derived from the renal cell carcinoma cell line deposited under accession number DSM ACC 2635 at the DSMZ.
8. A method of making a vaccine according to claim 1 for the treatment of patients who suffer from tumor diseases, wherein tumor cells of a genetically not identical (different) donor of the same species (allogeneic) are transfected ex-vivo with nucleic acid molecules encoding:
interleukin-7 (IL-7), granulocyte-macrophage colony stimulating factor (GM-CSF), CD40L/CD154 and B7.1/CD80
and subsequently are transferred into a an applicable pharmaceutical composition.
9. The method according to claim 8 , wherein:
additionally, an immune modulating oligodeoxyribonucleotide is employed as an adjuvant; and the immune modulating oligodeoxyribonucleotide comprises a circular strand of deoxyribonucleic acid with a partially complementary, antiparallel base sequence and is of a dumbbell shape.
10. The method according to claim 9 , wherein: the immune modulating oligodeoxyribonucleotide comprises a sequence of the base sequence N 1 N 2 CGN 3 N 4 , where N 1 N 2 is an element selected from the group comprising GT, GG, GA, AT or AA, N 3 N 4 is an element selected from the group comprising CT or TT, and C is desoxycytosine, G is desoxyguanosine, A is desoxyadenosine and T is desoxythymidine; and
the sequence with the base sequence N 1 N 2 CGN 3 N 4 is positioned in the single stranded region of the oligodeoxyribonucleotide and comprises 40 to 200 nucleotides.
11. The method according to claim 8 , wherein at least one of:
the nucleic acid molecules are present in one or several expression constructs; and
the allogeneic tumor cells are selected from colorectal carcinoma, small cell or non-small cell lung carcinoma, prostate carcinoma, mamma carcinoma, ovarian carcinoma, renal cell carcinoma and malign melanoma.
12. The method according to claim 8 , wherein the transfection method being ballistic transfer, polycation, transfection, calcium phosphate precipitation, microinjection, protoplast fusion, liposome fusion, viral transfection systems, lipofection and/or electroporation.
13. The method according to claim 9 , wherein the tumor cells are derived from several patients with the same disease picture.
14. The method according to claim 8 , wherein the tumor cells of the renal cell carcinoma cell line deposited under accession number DSM ACC 2635at the DSMZ are employed.
15. A human tumor cell transfected ex-vivo with nucleic acid molecules encoding:
interleukin-7 (IL-7), granulocyte-macrophage colony stimulating factor (GM-CSF), CD40L/CD154 and B7.1/CD80.
16. The human tumor cell according to claim 15 , wherein the expression construct is:
a) a plasmid; or
b) a linear double stranded, covalently closed expression cassette comprising essentially solely a CMV promoter, an intran, the encoding gene sequence and a polyadenylation sequence being closed on both ends of the double strand by a short loop of single stranded nucleotide residues.
17. The human tumor cell according to claim 16 , wherein said tumor cell is an allogeneic tumor cell of a renal cell carcinoma cell line.
18. The human tumor cell according to claim 17 , wherein the tumor cells are from the renal cell carcinoma cell line deposited under accession number DSM ACC 2635 at the DSMZ.
19. A composition comprising tumor cells of a genetically not identical (different) donor of the same species (allogeneic), wherein the tumor cells comprise ex-vivo-transfected, encoding nucleic acid molecules which encode:
interleukin-7 (IL-7), granulocyte-macrophage colony stimulating factor (GM-CSF), CD40L1CD154 and B7.1/CD80.
20. A human tumor cell for use as a vaccine in the treatment of tumor diseases comprising: tumor cells from the renal cell carcinoma cell line deposited under accession number DSM ACC 2635 at the DSMZ that has been transfected ex-vivo with nucleic acid molecules encoding:
interleukin-7 (IL-7), granulocyte-macrophage colony stimulating factor (GM-CSF), CD40L/CD154 and B7.1/CD80
and a corresponding expression construct, wherein the expression construct is a plasmid or a linear double stranded, covalently closed expression cassette comprising essentially solely a CMV promoter, an intron, the encoding gene sequence and a polyadenylation sequence being closed on both ends of the double strand by a short loop of single stranded nucleotide residues.