Engineered artificial antigen presenting cells for tumor infiltrating lymphocyte expansion
In some embodiments, compositions and methods relating to isolated artificial antigen presenting cells (aAPCs) are disclosed, including aAPCs comprising a myeloid cell transduced with one or more viral vectors, such as a MOLM-14 or a EM-3 myeloid cell, wherein the myeloid cell endogenously expresses HLA-AB/C, ICOS-L, and CD58, and wherein the one or more viral vectors comprise a nucleic acid encoding CD86 and a nucleic acid encoding 4-1BBL and/or OX40L and transduce the myeloid cell to express CD86 and 4-1BBL and/or OX40L proteins. In some embodiments, methods of expanding tumor infiltrating lymphocytes (TILs) with aAPCs and methods of treating cancers using TILs after expansion with aAPCs are also disclosed.
1. An isolated artificial antigen presenting cell (aAPC) comprising an EM-3 myeloid cell that endogenously expresses ICOS-L (inducible T-cell co-stimulator ligand), CD58, and one or more of HLA (human leukocyte antigen)-A, HLA-B, or HLA-C, wherein said aAPC is stably transduced with one or more viral vectors, wherein the one or more viral vectors comprise:
(i) a nucleic acid encoding CD86;
(ii) one or more nucleic acids encoding one or more costimulatory molecules selected from the group consisting of OX40L and 4-1BBL; and,
(iii) a nucleic acid encoding SEQ ID NO:27;
wherein the myeloid cell expresses on its surface a protein encoded by each of the nucleic acids of (i), (ii) and (iii).
2. The aAPC of claim 1 , wherein the aAPC can stimulate and expand tumor infiltrating lymphocytes (TILs) contacted with the aAPC.
3. The aAPC of claim 1 , wherein the aAPC expands a population of TILs by at least 50-fold over a period of 7 days in a cell culture medium comprising IL-2 (Interleukin-2) at a concentration of about 3000 IU/mL and OKT-3 antibody at a concentration of about 30 ng/mL.
4. The aAPC of claim 1 , wherein the aAPC can stimulate and expand a T cell contacted with the aAPC.
5. The aAPC of claim 1 , wherein the CD86 protein comprises a sequence as set forth in SEQ ID NO:8, or a sequence comprising one or more conservative amino acid substitutions thereof.
6. The aAPC of claim 1 , wherein the nucleic acid encoding CD86 comprises SEQ ID NO:19.
7. The aAPC of claim 1 , wherein the one or more costimulatory molecules comprises a 4-1BBL protein.
8. The aAPC of claim 7 , wherein the 4-1BBL protein comprises a sequence as set forth in SEQ ID NO:9, or a sequence comprising one or more conservative amino acid substitutions thereof.
9. The aAPC of claim 7 , wherein the one or more nucleic acids encoding the 4-1BBL protein comprises SEQ ID NO:16.
10. The aAPC of claim 1 , wherein the one or more costimulatory molecules comprises an OX40L protein.
11. The aAPC of claim 10 , wherein the OX40L protein comprises a sequence as set forth in SEQ ID NO:10, or a sequence comprising one or more conservative amino acid substitutions thereof.
12. An isolated artificial antigen presenting cell (aAPC) comprising an EM-3 cell that endogenously expresses ICOS-L (inducible T-cell co-stimulator ligand), CD58, and one or more of HLA-A, HLA-B, or HLA-C, wherein said aAPC is stably transduced with one or more viral vectors, wherein the one or more viral vectors comprise:
(i) a nucleic acid encoding CD86 or more conservative amino acid substitutions thereof;
(ii) one or more nucleic acids comprising a sequence encoding one or more amino acid sequence selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:13, and SEQ ID NO:14;
(iii) a nucleic acid encoding SEQ ID NO:27;
wherein the EM-3 cell expresses on its surface a protein encoded by each of the nucleic acids of (i), (ii) and (iii).