Formulations of PBMCs
The present application provides formulations of modified peripheral blood mononuclear cells (PBMCs), wherein the formulation comprises: PBMCs comprising at least one antigen (e.g., a human papillomavirus (HPV) antigen) and a cryopreservation medium. In some embodiments, the PBMCs are conditioned by incubating the PBMC in the presence of an adjuvant.
1. A pharmaceutical formulation comprising:
a) peripheral blood mononuclear cells (PBMCs), which comprise at least one antigen,
b) a cryopreservation medium,
c) a hypothermic preservation medium, which is present at a percentage of about 25% to about 35% (w/w), and
d) human serum albumin.
2. The pharmaceutical formulation of claim 1 , which comprises: (i) about 5×10 6 PBMCs to about 5×10 7 PBMCs; or (ii) about 1×10 6 PBMCs/mL to about 1×10 7 PBMC/mL.
3. The pharmaceutical formulation of claim 1 , wherein the cryopreservation medium is present in the formulation at a percentage of about 40% to about 95% (w/w).
4. The pharmaceutical formulation of claim 1 , wherein the hypothermic preservation medium comprises: (i) a water-soluble analog of vitamin E; (ii) trolox ((±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid); or (iii) both (i) and (ii).
5. The pharmaceutical formulation of claim 1 , wherein the human serum albumin: (i) is present in the formulation at a percentage of about 2% to about 8% (w/w) or (ii) is provided in a human serum albumin solution at a percentage of about 15% to about 25% (w/w).
6. The pharmaceutical formulation of claim 1 , wherein the pH of the formulation is about 6.0 to about 8.5.
7. The pharmaceutical formulation of claim 1 , which comprises:
a) about 1×10 6 PBMCs/mL to about 1×10 7 PBMCs/mL,
b) the cryopreservation medium at a percentage of about 65% to about 95% (w/w), and
c) the human serum albumin at a percentage of about 2% to about 8% (w/w),
wherein the pH of the formulation is about pH 6.0 to about pH 8.5.
8. The pharmaceutical formulation of claim 1 , which comprises:
a) about 1×10 6 PBMCs/mL to about 1×10 7 PBMCs/mL,
b) the cryopreservation medium at a percentage of about 40% to about 60% (w/w), and
c) the human serum albumin at a percentage of about 2% to about 8% (w/w), and
wherein the pH of the formulation is about pH 6.0 to about pH 8.5.
9. The pharmaceutical formulation of claim 1 , wherein the PBMCs comprise T cells, B cells, NK cells, monocytes, or combinations thereof.
10. The pharmaceutical formulation of claim 1 , wherein the at least one antigen comprises one or more human papillomavirus (HPV) antigens.
11. The pharmaceutical formulation of claim 10 , wherein the one or more HPV antigens comprise the amino acid sequence of any one of SEQ ID NOs: 1-4 and 18-25.
12. The pharmaceutical formulation of claim 1 , wherein the PBMCs have been conditioned with an adjuvant.
13. The pharmaceutical formulation of claim 12 , wherein the adjuvant comprises a CpG oligodeoxynucleotide (ODN), LPS, IFN-α, a STING agonist, a RIG-I agonist, poly I:C, R837, R848, a TLR3 agonist, a TLR4 agonist, a TLR 9 agonist, or a combination thereof.
14. A vial comprising the pharmaceutical formulation of claim 1 .
15. A method of producing a pharmaceutical formulation comprising PBMCs, the method comprising adding a cryopreservation medium, a hypothermic preservation medium, and human serum albumin to a population of PBMCs to produce the formulation, wherein the PBMCs comprise at least one antigen, and wherein the hypothermic preservation medium is added to a percentage of about 25% to about 35% (w/w).
16. The method of claim 15 , wherein
a) the population of PBMCs in the formulation is about 1×10 6 PBMCs to about 1×10 7 PBMCs,
b) the cryopreservation medium is added to a percentage of about 40% to about 60% (w/w), and/or
c)
the human serum albumin solution is added to a percentage of about 15% to about 25% (w/w), and
wherein the pH of the formulation is adjusted to about pH 6.0 to about pH 8.5.
17. The method of claim 15 , further comprising:
(a) passing a cell suspension comprising a population of input PBMCs through a cell-deforming constriction, thereby causing perturbations of the input PBMCs; and
(b) contacting the population of perturbed input PBMCs with the at least one antigen such that the at least one antigen passes through the perturbations and enters the perturbed input PBMCs to generate the population of PBMCs comprising the at least one antigen.
18. A method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject the pharmaceutical formulation of claim 1 .