Means and methods for influencing the stability of antibody producing cells
The invention provides a method for influencing the stability of an antibody producing cell, comprising directly or indirectly influencing the amount of BCL6 and/or Blimp 1 expression product within said antibody producing cell. Stable antibody producing cells and cell lines are also provided, as well as methods for producing antibodies using such cells and/or cell lines.
1. A method for producing an antibody producing cell culture that is able to replicate for at least one week, the method comprising:
providing a B cell;
increasing an expression level of Blimp-1 in said B cell by culturing the B cell in the presence of IL-21 and/or IL-10, or by providing the B cell with STAT3;
providing the B cell with a nucleic acid molecule encoding a peptide comprising BCL6; and
providing the B cell with a nucleic acid molecule encoding BCL-xL.
2. The method according to claim 1 , wherein the BOA expression level is brought to, and/or maintained at, essentially the same level or at a higher level as compared to a plasmablast.
3. The method according to claim 1 , wherein the B cell is provided with a nucleic acid molecule encoding BCL6, STAT3, Bcl-xL or a fusion protein of BCL6, regulated by an activator and/or repressor that is inducible by an exogenous compound.
4. The method according to claim 1 , comprising:
providing said B cell with Epstein Barr Virus; and
culturing said B cell.
5. The method according to claim 4 , comprising:
testing a plurality of B cells for a specificity for a given antigen;
selecting at least one B cell with a specificity for said given antigen;
providing said selected 13 cell with exogenous BCL6, or a fusion protein thereof;
selecting a B cell comprising exogenous BCL6, or a fusion protein thereof;
increasing an expression level of Blimp-1 in the B cell by culturing the B in the presence of IL-21 and/or IL-10, or by providing the antibody producing cell with STAT3; and
providing said selected B cell with Epstein Barr Virus.
6. The method according to claim 1 , wherein said B cell is cultured together with IL-21.
7. The method according to claim 1 , wherein said B cell is cultured together with IL-21 before BCL6, or a fusion protein thereof, is introduced into said B cell.
8. The method according to claim 1 , wherein an antibody producing B cell culture is produced that is able to replicate for at least six months.
9. The method according to claim 8 , wherein the B cell has been obtained from an individual, which individual had been previously exposed to the antigen of interest.
10. A method for producing an antibody producing cell culture able to replicate for at least one week, wherein the antibody producing cell expresses a level of Blimp-1, the method comprising:
increasing the expression level of Blimp-1 in the antibody producing cell by culturing the antibody producing cell in the presence of IL-21 and/or IL-10, or by providing the antibody producing cell with STAT3;
providing the antibody producing cell with a nucleic acid molecule encoding a peptide comprising BCL6; and
providing the antibody producing cell with a nucleic acid molecule encoding BCL-xL.
11. The method according to claim 10 , wherein the antibody producing cell is from a subject previously exposed to an antigen of interest.
12. The method according to claim 10 , further comprising:
providing an antibody producing cell with BCL6;
providing the antibody producing cell with Epstein Barr Virus; and
culturing the antibody producing cell.
13. The method according to claim 12 , comprising:
testing a plurality of antibody producing cells for a specificity for a given antigen;
selecting at least one antibody producing cell with a specificity for the given antigen;
providing the selected antibody producing cell with exogenous BCL6;
selecting an antibody producing cell comprising exogenous BCL6; and
providing the selected antibody producing cell with Epstein Barr Virus.
14. The method according to claim 12 , further comprising:
culturing the antibody producing cell together with IL-21.
15. The method according to claim 14 , wherein the antibody producing cell is first cultured together with IL-21 and afterwards providing BCL6 to the antibody producing cell.
16. The method according to claim 10 , wherein an antibody producing cell culture is produced that is able to replicate for at least six months.
17. A method for producing an antibody producing cell that is able to replicate for at least one week, the method comprising:
providing an antibody producing cell with a polynucleotide encoding a peptide comprising BCL6 to enhance the antibody producing cell's stability;
culturing the antibody producing cell with IL-21, IL-10, and/or STAT3 to increase expression of Blimp-1 in the antibody producing cell; and
providing the antibody producing cell with a polynucleotide encoding BCL-xL.
18. The method according to claim 1 , wherein the BCL6 forms a fusion protein.
19. The method according to claim 10 , wherein the BCL6 forms a fusion protein.
20. The method according to claim 17 , wherein the BCL6 forms a fusion protein.