US2019314412A1PendingUtilityA1

Methods and compositions high scale therapeutic production of memory nk cells

54
Assignee: UNIV CENTRAL FLORIDA RES FOUND INCPriority: Nov 8, 2016Filed: May 24, 2019Published: Oct 17, 2019
Est. expiryNov 8, 2036(~10.3 yrs left)· nominal 20-yr term from priority
Inventors:Alicja CopikRobert Y. IgarashiJeremiah OyerDeborah Altomare
A61P 31/12A61P 37/02A61P 35/00C12N 2501/2315C12N 2501/2318A61K 45/06A61K 38/2086A61K 35/33C12N 2501/20C12N 2501/2302A61K 38/20C12N 2501/2312C12N 2501/2321A61K 38/208C12N 5/0646A61K 35/17A61K 40/15A61K 2300/00A61K 2121/00A61K 48/0008A61K 48/005A61K 47/64
54
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

Disclosed are compositions and methods relating to the expansion of memory NK cells.

Claims

exact text as granted — not AI-modified
1 - 20 . (canceled) 
     
     
         21 . A method for increasing the number of memory NK cells comprising
 a) preactivating NK cells by contacting at least one NK cell with at least one stimulatory cytokine; and   b) expanding the preactivated NK cells of step a), by contacting the preactivated NK cells with plasma membrane (PM) particles with surface bound interleukin (IL)-21 (PM21) particles, exosomes (EX) with surface bound IL-21 (EX21) exosomes, or feeder cells (FC) with surface bound IL-21 (FC21) feeder cells, or any combination thereof.   
     
     
         22 . The method of  claim 21 , further comprising maintaining the contact in (b) for at least about 15 days, wherein a 10,000 fold increase in NK cell number is achieved. 
     
     
         23 . The method of  claim 21 , further comprising a washing step after the preactivation in step (a) and/or after the expansion in step (b). 
     
     
         24 . The method of  claim 21 , wherein the NK cells are obtained from an unselected population of peripheral blood mononuclear cells, cord blood or a stem cell population. 
     
     
         25 . The method of  claim 21 , wherein preactivating the NK cells comprises contacting the NK cells with at least one stimulatory cytokine selected from the group consisting of Interleukin-12 (IL-12), Interleukin-15 (IL-15), and Interleukin-18 (IL-18). 
     
     
         26 . The method of  claim 21 , wherein preactivating the NK cells comprises contacting the NK cells with any combination of two stimulatory cytokines selected from the group consisting of IL-12, IL-15, and IL-18. 
     
     
         27 . The method of  claim 21 , wherein preactivating the NK cells comprises contacting the NK cells with IL-12, IL-15, and IL-18. 
     
     
         28 . The method of  claim 24 , further comprising contacting the NK cells with an one or more NK cell effector agents selected from the group consisting of 4-1BB ligand (4-1BBL), Interleukin-2 (IL-2), Major histocompatibility complex class I-related chain AB (MICAS), UL16 Binding Protein 2 (ULBP2), Intracellular adhesion molecule 1(ICAM-1), 2B4, /signaling lymphocyte activation molecule (SLAM) family member 2 (F2) (BCM1/SLAMF2), cluster of differentiation 155 CD155), cluster of differentiation 112 (CD112), C-C chemokine receptor type 7 (CCR7), DnaX activation protein of 12 kDa (DAP12), and DnaX activation protein of 10 kDa (DAP10), and any combination thereof. 
     
     
         29 . The method of  claim 21 , wherein preactivating the NK cells comprises contacting the NK cells with the at least one stimulatory cytokine in vitro, in vivo, or ex vivo. 
     
     
         30 . The method of  claim 21 , wherein preactivating the NK cells comprises contacting the NK cells with at least one stimulatory cytokine for a period of about 6 to about 24 hours. 
     
     
         31 . The method of  claim 21 , wherein the PM21 particles, EX21 exosomes, or FC21 feeder cells comprise one or more stimulatory peptides coupled to a membrane-inserting peptide. 
     
     
         32 . The method of  claim 31 , wherein the stimulatory peptide coupled to a membrane-inserting peptide comprises a fused peptide that is capable of membrane insertion with affinity for a lipid bilayer, wherein the fused peptide comprises a segment of IG4, CD4, or a combination thereof. 
     
     
         33 . The method of  claim 31 , wherein the one or more stimulatory peptides coupled to a membrane-inserting peptide is a fusion protein encoded by recombinant DNA. 
     
     
         34 . The method of  claim 31 , wherein the membrane-inserting peptide comprises human Fc, GPI, trans-membrane T-cell receptor, or pHLIP. 
     
     
         35 . The method of  claim 31 , wherein the one or more stimulatory peptides are selected from the group consisting of 4-1BBL, IL-2, IL-12, IL-18, IL-21, MICA/B, ULBP2, ICAM-1, 2B4,BCM1/SLAMF2, CD155, CD112, CCR7, DAP12, and DAP10. 
     
     
         36 . The method of  claim 21 , wherein the NK cells are contacted with the PM21 particles, EX21 exosomes, or FC21 feeder cells in vitro, in vivo, or ex vivo. 
     
     
         37 . The method of  claim 21 , wherein the NK cells of step (a) are contacted with PM21 particles, EX21 exosomes, or FC21 feeder cells for a period of 7 days to 60 days. 
     
     
         38 . The method of  claim 21 , further comprising resting the expanded memory NK cells following step (b). 
     
     
         39 . The method of  claim 38 , wherein the cells are memory NK cells are rested for at least 1, 2, 3, 4 or 5 days. 
     
     
         40 . The method of  claim 38 , wherein the memory NK cells are rested for no more than 5 days. 
     
     
         41 . The method of  claim 21 , wherein the PM21 particles, EX21 exosomes, or FC21 feeder cells comprise 4-1BBL and IL-21. 
     
     
         42 . The method of  claim 41 , wherein the 4-1BBL and IL-21 are coupled to a membrane-inserting peptide. 
     
     
         43 . An activated NK cell composition comprising NK cells following contact with at least one stimulatory cytokine selected from the group consisting of IL-12, IL-15, and IL-18 for at least about 6 hours. 
     
     
         44 . The cell composition of  claim 43 , in contact with PM21 particles, EX21 exosomes, FC21 feeder cells, or any combination thereof. 
     
     
         45 . The composition of  claim 44 , further comprising an additional one or more cytokines selected from the group consisting of 4-1BBL, IL-2, IL-21, MICA/B, ULBP2, ICAM-1, 2B4BCM1/SLAMF2, CD155, CD112, CCR7, DAP12, DAP10, and any combination thereof. 
     
     
         46 . The composition of  claim 44 , in vitro or ex vivo. 
     
     
         47 . The composition of  claim 44 , wherein the PM21 particles, EX21 exosomes, or FC21 feeder cells comprise one or more stimulatory peptides coupled to a membrane-inserting peptide. 
     
     
         48 . The composition of  claim 47 , wherein each of the one or more stimulatory peptides coupled to a membrane-inserting peptide comprises a fused peptide that is capable of membrane insertion with affinity for a lipid bilayer, wherein the fused peptide comprises a segment of IG4, CD4, or a combination thereof. 
     
     
         49 . The composition of  claim 47 , wherein the one or more stimulatory cytokines coupled to a membrane-inserting peptide is a fusion protein encoded by recombinant DNA. 
     
     
         50 . The composition of  claim 47 , wherein the membrane-inserting peptide comprises human Fc, GPI, trans-membrane T-cell receptor, or pHLIP. 
     
     
         51 . The composition of  claim 47 , maintained in contact with the PM21 particles, EX21 exosomes, FC21 feeder cells or combination thereof for a period of 7 days to 60 days. 
     
     
         52 . The composition of  claim 47 , wherein the PM21 particles, EX21 exosomes, or FC21 feeder cells comprise IL-21 and further comprise 4-1BBL. 
     
     
         53 . The method of  claim 51 , wherein the 4-1BBL and IL-21 are coupled to a membrane-inserting peptide. 
     
     
         54 . A kit for increasing the number of memory NK cells comprising amounts of one or more stimulatory cytokines and amounts of one or more vesicles comprising IL-21, and optionally amounts of one or more NK cell effector agents. 
     
     
         55 . The kit of  claim 54 , wherein the one or more stimulatory cytokines are selected from the group consisting of IL-12, IL-15, IL-18 and any combination thereof. 
     
     
         56 . The kit of  claim 54 , wherein the one or more vesicles comprising IL-21 are selected from a PM21 particle, an EX21 exosome, a FC21 feeder cell and any combination thereof. 
     
     
         57 . The kit of  claim 54 , wherein the one or more NK cell effector agents are selected from the group consisting of 41BBL, IL-2, IL-15, IL-21, MICA/B, ULBP2, ICAM-1, 2B4, BCM1/SLAMF2, CD155, CD112, CCR7, DAP12, DAP10 and any combination thereof. 
     
     
         58 . The kit of  claim 54 , further comprising an amount of NK cells or an NK cell line.

Join the waitlist — get patent alerts

Track US2019314412A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.