Compositions and methods for modified B cells expressing reassigned biological agents
Compositions and methods are disclosed herein for producing one or more immunoglobulins in an isolated cytotoxic B lymphocyte cell line. An isolated cell line includes an isolated B lymphocyte cell line capable of expressing at least one exogenously incorporated membrane immunoglobulin capable of binding to a first antigen and at least one endogenous secreted immunoglobulin capable of binding to a second antigen, and further capable of expressing at least one exogenously incorporated recombinant B cell receptor that signals for expression of cytotoxic effector molecules.
1. A modified B cell comprising:
at least one exogenous nucleic acid encoding a membrane immunoglobulin or a recombinant B cell receptor that is capable of binding to a first antigen;
at least one exogenous nucleic acid encoding a reassigned biological agent; and
at least one endogenous secreted immunoglobulin reactive to a second antigen.
2. The modified B cell of claim 1 , wherein the membrane immunoglobulin includes one or more exogenous membrane immunoglobulin polypeptides.
3. The modified B cell of claim 1 , wherein the exogenously incorporated exogenous nucleic acid encoding the at least one membrane immunoglobulin is integrated in one or more chromosomal loci.
4. The modified B cell of claim 3 , wherein the chromosomal loci include at least one of the Ig H chain or Ig L chain chromosomal loci.
5. The modified B cell of claim 4 , wherein the Ig H chain or Ig L chain chromosomal loci are located on the non-expressed Ig allele.
6. The modified B cell of claim 5 , wherein at least one of the Ig H chain or Ig L chain chromosomal loci is under expression control of an endogenous promoter or enhancer elements.
7. The modified B cell of claim 5 , wherein the Ig L chain locus includes at least one of the kappa or lambda light (L)-chain locus.
8. The modified B cell of claim 7 , wherein the integration at the kappa or lambda light (L)-chain locus disrupts endogenous B cell kappa or lambda light (L)-chain expression.
9. The modified B cell of claim 3 , wherein the chromosomal loci include one or more non-Ig L or non-Ig H chromosomal loci in the B lymphocyte cell line.
10. The modified B cell of claim 3 , wherein the at least one exogenous nucleic acid includes an exogenous bicistronic expression construct.
11. The modified B cell of claim 1 , wherein the at least one exogenously incorporated nucleic acid encoding the membrane immunoglobulin includes a nucleic acid encoding one single chain Fv immunoglobulin.
12. The modified B cell of claim 1 , wherein the modified B cell includes at least one of a nave B lymphocyte, immature B lymphocyte, transitional B lymphocyte, mature B lymphocyte, B1 B lymphocyte, marginal zone B lymphocyte, follicular B lymphocyte, memory B lymphocyte, plasmablast, or plasma cell.
13. The modified B cell of claim 1 , wherein the modified B cell is part of a polyclonal population of B lymphocytes.
14. The modified B cell of claim 1 , wherein the modified B cell is part of a monoclonal population of B lymphocytes.
15. The modified B cell of claim 1 , wherein the modified B cell is part of a modified B lymphocyte cell line.
16. The modified B cell of claim 1 , wherein the membrane immunoglobulin includes at least one of a membrane anchor, a cytoplasmic domain, a hinge region or an extracellular ligand-binding domain.
17. The modified B cell of claim 1 , wherein the modified B cell bears at least one chimeric B cell receptor capable of transducing signals for inducing expression of cytotoxic effector molecules.
18. The modified B cell of claim 17 , wherein the chimeric B cell receptor is derived from at least one of membrane IgG, CD19, BCMA, CD38, the common gamma chain receptor, the IL-21 receptor, Toll-like Receptor, or CD40.
19. The modified B cell of claim 17 , wherein the cytotoxic effector molecules include at least one of perforin, granzyme b, Fas ligand, or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).
20. The modified B cell of claim 17 , wherein the cytotoxic effector molecules are expressed in response to a specific target cell or target antigen.
21. A modified B cell comprising:
at least one exogenous nucleic acid encoding a membrane immunoglobulin or recombinant B cell receptor that is capable of binding to a first antigen;
at least one exogenous nucleic acid encoding a reassigned biological agent; and
at least one exogenous nucleic acid encoding secreted immunoglobulin capable of binding to a second antigen.
22. The modified B cell of claim 21 , wherein the at least one exogenous nucleic acid encoding the membrane immunoglobulin includes at least one exogenous membrane immunoglobulin polypeptide.
23. The modified B cell of claim 21 , wherein the exogenous nucleic acid encoding the membrane immunoglobulin is integrated in one or more chromosomal loci in the modified B cell.
24. The modified B of claim 23 , wherein the chromosomal loci include at least one of the Ig H chain or Ig L chain chromosomal loci.
25. The modified B cell of claim 24 , wherein the Ig H chain or Ig L chain chromosomal loci are non-expressed Ig alleles.
26. The modified B cell of claim 24 , wherein the Ig H chain or Ig L chain chromosomal loci are expressed Ig alleles.
27. The modified B cell of claim 26 , wherein at least one of the Ig H chain or Ig L chain chromosomal loci is under expression control of endogenous promoter or enhancer elements.
28. The modified B cell of claim 26 , wherein the Ig L chain locus includes at least one of the kappa or the lambda light (L)-chain locus.
29. The modified B cell of claim 28 , wherein the integration at the kappa or lambda light (L)-chain locus disrupts endogenous B cell kappa or lambda light (L)-chain expression.
30. The modified B cell of claim 24 , wherein the at least one exogenous nucleic acid encoding the membrane immunoglobulin includes an exogenous bicistronic expression construct.
31. The modified B cell of claim 21 , wherein the at least one exogenous nucleic acid encoding the membrane immunoglobulin includes a nucleic acid encoding one single chain Fv immunoglobulin.
32. The modified B cell of claim 21 , wherein the modified B cell includes at least one of nave B lymphocyte, immature B lymphocyte, transitional B lymphocyte, mature B lymphocyte, B1 B lymphocyte, marginal zone B lymphocyte, follicular B lymphocyte, memory B lymphocyte, plasmablast, or plasma cell.
33. The modified B cell of claim 21 , wherein the membrane immunoglobulin includes at least one of a membrane anchor, a cytoplasmic domain, a hinge region, or an extracellular ligand-binding domain.
34. The modified B cell of claim 21 , wherein the modified B cell bears at least one chimeric B cell receptor capable of transducing signals for inducing expression of cytotoxic effector molecules.
35. The modified B cell of claim 34 , wherein the chimeric B cell receptor is derived from at least one of membrane IgG, CD19, BCMA, CD38, the common gamma chain receptor, the IL-21 receptor, Toll-like Receptor, or CD40.
36. The modified B cell of claim 35 , wherein the cytotoxic effector molecules include at least one of perforin, granzyme B, Fas ligand, or tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).
37. The modified B cell of claim 35 , wherein the cytotoxic effector molecules are expressed in response to a specific target cell or target antigen.
38. The modified B cell of claim 21 , wherein the at least one exogenous nucleic acid encoding secreted immunoglobulin includes at least one exogenous secreted immunoglobulin polypeptide.
39. The modified B cell of claim 38 , wherein the at least one exogenous nucleic acid encoding secreted immunoglobulin is encoded by at least one exogenous nucleic acid.
40. The modified B cell of claim 39 , wherein the exogenous nucleic acid encoding secreted immunoglobulin is integrated in one or more chromosomal loci in the modified B cell.
41. The modified B cell of claim 40 , wherein the chromosomal loci include at least one of the Ig H chain or Ig L chain chromosomal loci.
42. The modified B cell of claim 41 , wherein the Ig H chain or Ig L chain chromosomal loci are the non-expressed Ig alleles.
43. The modified B cell of claim 40 , wherein the Ig H chain or Ig L chain chromosomal loci are the expressed Ig alleles.
44. The modified B cell of claim 43 , wherein at least one of the Ig H chain or Ig L chain chromosomal loci is under expression control of endogenous promoter or enhancer elements.
45. The modified B cell of claim 40 , wherein the at least one exogenous nucleic acid encoding a secreted immunoglobulin includes an exogenous bicistronic expression construct.
46. The modified B cell of claim 40 , wherein the at least one exogenous nucleic acid encoding a secreted immunoglobulin includes a nucleic acid encoding one single chain FIT immunoglobulin.
47. A modified B cell comprising: at least one exogenous nucleic acid encoding a membrane immunoglobulin or a recombinant B cell receptor that is reactive to a first antigen;
at least one endogenous secreted immunoglobulin reactive to a second antigen;
at least one exogenous nucleic acid encoding a reassigned biological agent;
and at least one exogenous nucleic acid encoding a membrane receptor configured to directly or indirectly induce expression of at least one cytotoxic effector molecule.
48. The modified B cell of claim 47 , wherein the membrane receptor configured to directly or indirectly induce at least one cytotoxic effector molecule includes at least one chimeric receptor exhibiting at least a portion of at least one cytokine receptor.