Method for determining the immunogenicity of an antigen
The present invention relates to methods for preparing an artificial immune system. The artificial immune system comprises a cell culture comprising T cells, B cells and antigen-primed dendritic cells. The artificial immune system of the present invention can be used for in vitro testing of vaccines, adjuvants, immunotherapy candidates, cosmetics, drugs, biologics and other chemicals.
1. A method for immortalizing B cells monoclonal for an antigen, comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b);
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells;
(f) identifying B cells in the culture of (e) that are monoclonal for the antigen; and
(g) isolating and immortalizing B cells identified in (f) that are monoclonal for the antigen.
2. The method of claim 1 , wherein the serum-free cell culture media is X-VIVO 15.
3. The method of claim 1 , further comprising adding the antigen to the cell culture (e).
4. A method for producing antibodies having binding specificity for an antigen, comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b); and
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells.
5. The method of claim 4 , further comprising isolating antibodies specific for the antigen from the culture of (e).
6. The method of claim 4 , wherein the serum-free cell culture media is X-VIVO 15.
7. The method of claim 4 , further comprising adding the antigen to the cell culture (e).
8. A method for immortalizing B cells producing antibodies having binding specificity for an antigen, comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free cell culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b);
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells;
(f) identifying B cells producing antibodies having binding specificity for the antigen in the culture of (e); and
(g) isolating and immortalizing B cells identified in (f) producing antibodies having binding specificity for the antigen.
9. The method of claim 8 , wherein the serum-free cell culture media is X-VIVO 15.
10. The method of claim 8 , further comprising adding the antigen to the cell culture of (e).
11. A method for producing a therapeutic composition comprising monoclonal antibodies having binding specificity for an antigen, the method comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free cell culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b);
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells;
(f) identifying B cells in the culture of (e) that are monoclonal for the antigen;
(g) isolating and immortalizing B cells identified in (f) that are monoclonal for the antigen; and
(h) preparing a therapeutic composition comprising monoclonal antibodies produced by the immortalized B cells of (g) and a pharmaceutically acceptable carrier.
12. A method for producing a therapeutic composition comprising a monoclonal antibody having binding specificity for an antigen, the method comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free cell culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b);
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells;
(f) identifying B cells in the culture of (e) that are monoclonal for the antigen;
(g) isolating and immortalizing B cells identified in (f) that are monoclonal for the antigen; and
(h) preparing a therapeutic composition comprising monoclonal antibodies produced by one of the immortalized B cells of (g) and a pharmaceutically acceptable carrier.
13. A method for producing a therapeutic composition comprising antibodies having binding specificity for an antigen, the method comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free cell culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b);
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells;
(f) identifying B cells in the culture of (e) producing antibodies having binding specificity for the antigen;
(g) isolating and immortalizing B cells identified in (f) having binding specificity for the antigen; and
(h) preparing a therapeutic composition comprising antibodies having binding specificity for the antigen produced by the immortalized B cells of (g) and a pharmaceutically acceptable carrier.
14. A method for preparing a therapeutic composition comprising antibodies having binding specificity for an antigen, the method comprising:
(a) priming a population of dendritic cells with an antigen;
(b) adding the antigen-primed dendritic cells of (a) to a cell culture comprising T cells and serum-free cell culture media;
(c) priming a population of B cells with the antigen;
(d) adding the antigen-primed B cells of (c) to the cell culture comprising T cells and antigen-primed dendritic cells of (b);
(e) culturing the cell culture of (d) under conditions promoting production of antibodies by the antigen-primed B cells;
(f) isolating antibodies specific for the antigen from the culture of (e); and
(g) preparing a therapeutic composition comprising the isolated antibodies of (f) and a pharmaceutically acceptable carrier.