Method for inducing and accelerating cells
The present invention relates to a method for the production of functional dendritic cells wherein CD34 positive cells are contacted with compounds inducing and accelerating the differentiation of these CD34 positive cells into functional dendritic cells. More in particular, the CD34 positive cells are contacted with anthracyclines and/or anthracenediones. In another aspect, the current invention relates to the cells obtainable by the method according to the invention. In a further aspect the current invention relates to the use of compounds such as anthracyclines and/or anthracenediones that induce and accelerate the differentiation of CD34 positive cells into functional dendritic cells in the manufacture of a medicament for inducing an immune response in human in need thereof.
1. A method for the production of dendritic cells, the method comprising:
contacting CD34 positive cells with an anthracycline and/or an anthracenedione in a culture medium thereby increasing the rate of production of dendritic cells from the CD34 positive cells in comparison to CD34 positive cells not contacted with anthracycline and/or anthracenedione;
allowing the CD34 positive cells to develop into dendritic cells; and
harvesting dendritic cells.
2. The method according to claim 1 , wherein the anthracycline and/or an anthracenedione is selected from the group consisting of daunorubicin, doxorubicin, pirarubicin, aclarubicin, epirubicin, oxaunomycin, andidarubicin and mitoxantrone.
3. The method according to claim 1 , wherein the CD34 positive cells are contacted with the anthracycline and/or an anthracenedione for a period of between 1 and 7 days.
4. The method according to claim 3 , wherein the CD34 positive cells are contacted with the anthracycline and/or an anthracenedione for a period of between 2 and 4 days.
5. The method according to claim 1 , further comprising contacting the cells with at least one additional compound able to induce differentiation of the CD34 positive cells into dendritic cells.
6. The method according to claim 5 , wherein the at least one additional compound that is able to induce differentiation of the CD34 positive cells into dendritic cells is selected from the group consisting of GM-CSF, TNF-alpha, IL-4, and TGF-beta 1.
7. The method according to claim 1 , wherein at least one compound able to induce maturation of dendritic cells is present in the culture medium.
8. The method according to claim 7 , wherein the compound which is able to induce maturation of dendritic cells is selected from the group consisting of TNF-alpha, IL-6, PGE2 and IL-1Beta.
9. The method according to claim 1 , wherein the dendritic cells are selected from the group consisting of interstitial dendritic cells, immature dendritic cells, Langerhans dendritic cells, plasmatoid dendritic cells, and mature dendritic cells.
10. The method according to claim 1 , wherein the CD34 positive cells are MUTZ3 cells, human cells, or human tumor cells.
11. The method according to claim 10 , wherein MUTZ3 cells are contacted with from 0.05 nM to 20 nM mitoxantrone and/or from 10 to 120 nM doxorubicin, in the presence of from 50 to 150 ng/ml GM-CSF, from 5 to 20 ng/ml IL-4 and from 0.5 to 4 ng/ml TNF-alpha.
12. The method according to claim 10 , wherein MUTZ3 cells are contacted with from 0.05 nM to 20 nM mitoxantrone and/or from 10 to 120 nM doxorubicin, in the presence of from 5 to 20 ng/ml TGF-beta 1, from 50 to 150 ng/ml GM-CSF, and from 0.5 to 4 ng/ml TNF-alpha.
13. The method according to claim 1 , wherein said dendritic cells have an increased expression of HLA-DR.
14. The method comprising according to claim 1 , further comprising:
administering the harvested dendritic cells to a subject.