DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
The instant application, filed on 28 August 2023, claims domestic benefit to US provisional application no. 63/152,787, filed on 02/23/2021 and PCT/US2022/017526 filed on 02/23/2022.
Status of Application, Amendments, and/or Claims
The response filed on 18 June, 2026 has been entered in full. The response is an amendment to a Non-Final Rejection to the claim set filed on 25 July, 2025. In the amendment claims 1-3, 5, 24, 30, 33, 40, and 41 are amended, claims 27, 29, 42, 44, 48, 49, and 51 are previously presented, claims 55-57 are new, and claims 4, 6-23, 25, 26, 28, 31, 32, 34-39, 45-47, 50, and 52-54 are cancelled. Therefore, claims 1-3, 5, 24, 27, 29, 30, 33, 40-42, 44, 48-51, and 55-57 are pending and are the subject of this office action.
Status of Objections and Rejections
In the office action of 08/13/2026
Claims 7 and 10 were rejected under 35 U.S.C 102(a)(1) over Safina et at. The cancellation of the claims has rendered the rejections moot and they are withdrawn.
Claim 15 was rejected under 35 U.S.C. 102(a)(1) and 35 U.S.C. 102(a)(2) over Scott et al. The cancellation of the claim has rendered the rejection moot and it is withdrawn.
Claim 31 was rejected under 35 U.S.C 112(b) and 35 U.S.C. 103 over Safinia et al. in view of Marek et al. The cancellation of the claim has rendered the rejections moot and they are withdrawn.
Claim 30 was rejected under 35 U.S.C. 112(b). The amendment of the claim has overcome this rejection.
Claims 1-3, 48, and 49 were rejected under 35 U.S.C. 102(a)(1) over Safinia et al., the amendment to claim 1 has necessitated a new 35 U.S.C. 103 rejection over Safinia et al. in view of Yu et al. and Hippen et al.
Claims 24, 27, 40, and 51 were rejected under 35 U.S.C. 103 over Safinia et al. in view of McDonald et al. The amendment to claim 1 has necessitated a new 35 U.S.C. 103 rejection over Safinia et al. in view of Yu et al. and Hippen et al. and further in view of McDonald et al.
Claims 5, 30, and 41 were rejected under 35 U.S.C. 103 over Safinia et al. in view of Ye et al. and Hippen et al. The rejection is maintained.
Claim 29 was rejected under 35 U.S.C. 103 over Safinia et al. in view of Frost et al. The amendment to claim 1 has necessitated a new 35 U.S.C. 103 rejection over Safinia et al. in view of Yu et al. and Hippen et al. and further in view of Frost et al.
Claim 42 was rejected under 35 U.S.C. 103 over Safinia et al. in view of Marek et al. The amendment to claim 1 has necessitated a new 35 U.S.C. 103 rejection over Safinia et al. in view of Yu et al. and Hippen et al. and further in view of Marek et al.
Claim 33 is rejected under 35 U.S.C. 103 over Safinia et al. in view of Osborn et al. The amendment to claim 1 has necessitated a new 35 U.S.C. 103 rejection over Safinia et al. in view of Yu et al. and Hippen et al. and further in view of Osborn et al.
Claim 44 was rejected under 3 U.S.C. 103 over Safinia et al. in view of Marek-Trzonkowska. The amendment to claim 1 has necessitated a new 35 U.S.C. 103 rejection over Safinia et al. in view of Yu et al. and Hippen et al. and further in view of Marek-Trzonkowska.
Specification
The use of the term DSORTTM, which is mark used in commerce and further does not seem to registered as a trade name, has been noted in this application.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 1-3, 5, 7, 10, 30, 41, 48, 49, and 55 are rejected under 35 U.S.C. 103 as being unpatentable over Safinia et al. (2016) (Of Record, IDS 04/12/2024) in view of Yu et al. (2019) "Optimization of human T cell activation and expansion protocols improves efficiency of genetic Modification and overall cell yield." European Journal of Immunology (Vol. 49) (hereafter Yu) and Hippen et al. (2011) Massive ex Vivo Expansion of Human Natural Regulatory T cells (Tregs) with Minimal Loss of in Vivo Functional Activity Sci. Transl. Med. (3) 83ra41-83ra41 (hereafter Hippen).
In regard to claims 1 and 48 Safinia teaches a method for producing an expanded Treg cell population with a first stimulating step of anti-CD3/CD28 coated beads. Safinia further teaches a resting step of 4 days before fresh beads are added to the culture again (pg.7573, col 2, lines 7-23), followed by a restimulation step 12 days after an initial stimulation step (pg.7573, col 2, lines 7-23).
In regard to claim 2 Safinia teaches a method for expanding T reg cells which further comprises a 4-day rest period after restimulation, followed by the addition of new anti-CD3/CD28 beads to the culture (pg.7573, col 2, lines 7-23).
In regard to claim 3 Safinia teaches a method for expanding T reg cells which further comprises a third stimulation step 24 days after the first restimulation step and 12 days after a second stimulation step preceded by a 4-day rest period after which new anti-CD3/CD28 beads were added to the culture. Further after the stimulation the cells were harvested and anti-CD3/CD28 beads were removed from the cells (third resting step) for further processing (pg.7573, col 2, lines 7-23).
In regard to claim 49 Safinia teaches the expended Treg population as outlined above for claim 1, further Safinia teaches the cryopreservation of the Treg cells (pg.7574, col 2, lines 6-13).
Safinia fails to teach the genetic engineering of the second rested population of claim 5, the 1–5 day stimulations and 1-5 days’ rest before genetic modification of claims 1-3 and 30, and the 1000-fold increase in 11 days of claim 41.
Yu, however, in regard to claim 55 teaches a method of expanding Treg cells for 1-3 days, followed by genetic engineering with a lentivirus or CRISPR Cas 9 (Figure 2) for improved knockout efficiency (abstract).
Further, in regard to claim 41 Yu teaches a method in which one stimulation step for 3 days followed by significant dilution of the cell culture and maintenance of a low concentration of cells resulted in more than 1000-fold expansion of T-cells over 10-12 days (abstract)
Yu fails to teach the multiple stimulations and rest periods of claims 5 and 30.
Hippen, however teaches that the restimulation of Treg cells after letting them return to a resting condition greatly expanded the Treg expansion (pg.3, col 1, lines 13-37) and further teaches maximizing yield to Treg cells is critical for efficiency and reproducibility in graft vs host disease suppression (pg. 1, col 1, lines 12-14).
Thus, Safinia discloses a method of expanding T reg cells comprising up to 3 stimulation steps and 3 rest steps, and Yu teaches genetic engineering after 1–3-day expansion and maintaining a low cell concentration for a more than 1000 -fold expansion of the T-cells. Further Hippen teaches that restimulation of resting Treg cells expanded Treg expansion. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Safinia, Yu, and Hippen because keeping cells at low concentrations after stimulation and further restimulation allows for increased Treg cell expansion to yield more modified Treg cells and genetic engineering after stimulation allows for a better knockout efficiency to further yield more therapeutic product for use. Thus, combining the references would allow, with a reasonable expectation of success, the development of a method of Treg cell expansion and further genetic modification to yield a larger population of modified Treg cells with higher therapeutic efficacy.
Claims 24, 27, 40, and 51 are rejected under 35 U.S.C. 103 as being unpatentable over Safinia in view of Yu and Hippen as applied to claims 1-3 above, and further in view of McDonald et al. (2019) (of record, IDS 02/07/2025).
In regard to claims 24, 27, 40, and 51 Safinia in view of Yu and Hippen teaches the method of expansion of Treg cells using up to three stimulating steps and up to three rest steps as outlined above in the 35 U.S.C. 103 rejection for claims 1-3. Also noted in regard to claim 24 Safinia teaches the use of an antibody complex for stimulating step in the form of anti-CD3/CD28 supermagnetic beads (option C of the claim) (pg.7573, col 2, lines 7-23). Further, in regard to claim 27 Safinia teaches a method which does not use an artificial antigen presenting cell.
Safinia in view of Yu and Hippen fails to teach the antibody complex which can specifically bind to a combination of CD3, CD28, CD2 or the antibody or antigen binding fragment of anti-CD3 and/or anti-CD28 of claim 24. Safinia in view of Yu and Hippen also fails to teach a method which does not use supermagnetic beads of claim 27 or a method which does not use rapamycin of claims 27 and 40. Safinia in view of Yu and Hippen, further, fails to teach a method of treating an autoimmune or inflammatory disease, treating or preventing graft vs host disease or decreasing an immune response using an effective amount of Tregs of claim 51.
MacDonald, however, in regard to claim 24 teaches the use of a variety of methods to expand Treg cells (Table 1) which include an anti-CD3 monoclonal antibody attached to an artificial presenting cell which is more effective at expanding umbilical cord blood Tregs than supermagnetic beads (pg.56, col 1, lines 9-15). Further MacDonalds teaches that Stemcell Technologies has developed an antibody complex which can specifically bind to a combination of CD3 and CD28 or CD3, CD28, and CD2 to provide an activation reagent which is soluble and make their removal from culture easier (pg.56, col 1, lines 1-2 and col 2, lines 6-10).
In regards to claims 27 MacDonald teaches the success of various methods to Treg expansion which include method which do not use supermagnetic beads due to their need to be removed before infusion (pg.56, col 1, lines 8-11), and methods that do not use artificial antigen presenting cells due to their complication to the cell manufacturing process (pg.56, col 1, lines 26-29).
In regard to claims 27 and 40 MacDonald teaches methods which do not use rapamycin because it limits Treg proliferation (Table 1).
In regard to claim 51 MacDonald teaches it is commonly accepted, and clinical trials have been conducted to prove its safety to use adoptive transfer of regulatory T cells for preventing or treating graft vs host disease and autoimmune conditions (pg.52, col 1, lines 1-17). Further MacDonald teaches adoptive transfer of Treg cells works by shifting the balance of Tregs and T effector cells to favor tolerance induction (decreasing an immune response) (pg.52, col 1, lines 7-10).
Thus, Safinia in view of Yu and Hippen discloses a method of expanding Treg cells in which the cells are stimulated and rested up to three times for 1-5 days using anti-CD3/CD28 supermagnetic beads, and MacDonald teaches various expansion methods which includes using monoclonal antibody attached to artificial antigen presenting cells or antibody complexes for varying efficiency and post stimulation steps. Further MacDonald teaches rapamycin omission to support Treg cell proliferation and the use of expanded Tregs to decrease immune response, treat autoimmune conditions, or treat or prevent graft vs host disease. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Safinia in view of Yu and Hippen with the teachings of MacDonald because omission of rapamycin to the culture of the Treg cells can be used to balance Treg expansion size for better control of the Treg product for administration. Further MacDonald motivates the use of the platform taught in Safinia in view of Yu and Hippen for the treatment of graft vs host disease and autoimmune conditions. Thus, combining the references would allow, with a reasonable expectation of success, the development of a method to expand Treg cells which can have with more control on the purity and amount harvested of the end product for use in treating graft vs host disease and autoimmune conditions.
Claim 29 is rejected under 35 U.S.C. 103 as being unpatentable over Safinia in view of Yu and Hippen as applied to claim 1 above, and further in view of Frost et al. (US20190367876, of record IDS 02/07/2025).
The method taught in Safinia teaches culturing the cells in the presence of IL-2. Safinia in view of Yu and Hippen fails to teach the culturing of N-Acetyl-L-cysteine (NAC), however, frost teaches the addition of NAC in the culture when expanding immune cells including T-cells (pg.11, col 2, lines 1-3). Frost, further, teaches NAC was found to be beneficial during Treg expansion but detrimental during Treg cell modification (pg.11, col 2, lines 1-3).
Thus, Safinia in view of Yu and Hippen discloses a method of expanding T reg cells comprising at least a 1st stimulation step, a 1st rest step and a 2nd stimulation step for 1-5 days each, and Frost teaches the addition of NAC as a benefit for the expansion of Treg cells. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Safinia in view of Yu and Hippen with the teachings of Frost because the addition of NAC to the culture of expanding Treg cells increases expansion to yield a larger number of modified Treg cells. Thus, combining the references would allow, with a reasonable expectation of success, the development of a method for Treg cell expansion comprising a stimulation step and a rest step and wherein the cells are cultured with NAC for increased yield of modified Treg cells for use as a therapeutic.
Claim 42 is rejected under 35 U.S.C. 103 as being unpatentable over Safinia in view of Yu and Hippen as applied to claim 1 above, and further in view of Marek et al. (2011) The Time Is Crucial for Ex Vivo Expansion of T Regulatory Cells for Therapy Cell Transplantation (20) 1747-1758 (hereafter Marek).
Safinia in view of Yu and Hippen also fails to teach the method resulting in Tregs with a smaller surface area than Tregs that are cultured in the presence of a stimulating agent for 6 days; and/or wherein the method results in an increased proportion of Helios+Foxp3+ Tregs as compared to Tregs that are cultured in the presence of a stimulating agent for 6 days; and/or wherein the method results in Tregs with an increased ability to suppress proliferation of effector T cells (Teffs) as compared to Tregs that are cultured in the presence of a stimulating agent for 6 days; and/or wherein the method prevents overstimulation of the population of Tregs; and/or wherein the method reduces activation- induced cell death as compared to Tregs that are cultured in the presence of a stimulating agent for 6 days of claim 42.
Marek however teaches that suppressive phenotype and suppressive function decrease with time and that the length of culture was found to be a crucial variable to keep the cells regulatory (pg.1751, col 2, lines 1-6).
Thus, Safinia, in view of Yu and Hippen discloses a method of expanding T reg cells comprising at least a 1st stimulation step, a 1st rest step and a 2nd stimulation step for 1-5 days each, and Marek teaches the importance of minimizing timing in T cell expansion to maintain cellular suppressive function and viability. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to try the teachings of Safinia informed by the teachings of Marek because longer stimulation times induce cell death and degrades the Treg suppressive function. Thus, combining the references would allow, with a reasonable expectation of success, the development of a method of expanding Treg cells comprising a stimulation step and a rest step in which the time of stimulation steps are shortened to preserve Treg cell suppression function for a more efficacious therapeutic.
Claim 33 is rejected under 35 U.S.C. 103 as being unpatentable over Safinia in view of Yu and Hippen as applied to claim 1 above, and further in view of Osborn et al. US2021/0213062 (priority date 05/17/2018).
Safinia in view of Yu and Hippen fails to teach Treg cells expanded about 250-fold change before genetic engineering, however Osborn teaches expanded cells can be multiplied by 200-fold or 300-fold and any and all whole or partial integers there between (pg.33, col 2, lines 16-23). Further, Osborn teaches subsequent electroporation to introduce a nucleic acid encoding an agent (pg.34, col 1, lines 61-65).
Thus, Safinia discloses a method of expanding T reg cells comprising at least a 1st stimulation step, a 1st rest step and a 2nd stimulation step for about 1-5 days each and Osborn teaches a method wherein the Treg cells can be expanded to about 250 before genetic modification. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to try the combination of the teachings of Safinia with the teachings of Osborn because 250-fold expansion of Tregs is sufficient to allow for the genetic modification of Tregs. Thus, combining the references would allow, with a reasonable expectation of success, the development of a method of expanding Treg cells before genetic modification in which the cells are expanded to 250-fold to yield a sufficient number of Treg cells for genetic modification.
Claim 44 is rejected under 35 U.S.C. 103 as being unpatentable over Safinia in view of Yu and Hippen as applied to claim 1 above, and further in view of Marek-Trzonkowska (2017) (of record IDS 04/17/2024).
Safinia in view of Yu and Hippen fails to teach the method wherein at least 75% of Helios expression in the Tregs is maintained, the Tregs are Helios+ and/or have a fully demethylated Treg-Specific region (TSDR). Marek-Trzonkowska, however, teaches that when Treg cells are expanded at both 33°C and 37°C they maintained or increased their IKZF2 (Helios) expression (Figure 4). Further, Marek-Trzonkowska teaches that Treg cells expanded at 33°C showed significantly higher frequencies of demethylated TSDR than Treg expanded at 37°C, with both conditions resulting in close to 100% of cells having demethylated TSDRs (pg.5 lines 14-17 /Figure 3D).
Thus, Safinia, in view of Yu and Hippen discloses a method of expanding T reg cells comprising at least a 1st stimulation step, a 1st rest step and a 2nd stimulation step for about 1-5 days each, and Marek-Trzonkowska teaches culture at 37°C and even more so at 33°C maintain Helios expression and have demethylated TSDRs. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Safinia with the teachings to Marek-Trzonkowska because temperatures of 37°C or 33°C maintain Helios expression and demethylate the TSDRs for maintained Treg function. Thus, combining the references would allow, with a reasonable expectation of success, the development of a method of expanding Treg cells comprising a stimulation step and a rest step in which the Treg cells are cultured at 37°C or 33°C for maintenance of Helios expression and a fully demethylated TSDR for Treg function maintenance to maintain their use as a therapeutic.
Claims 56 and 57 are rejected under 35 U.S.C. 103 as being unpatentable over Safinia in view of Yu and Hippen as applied to claims 1 and 55 above, and further in view of Rosenblum et al. (2016) Regulatory T cell memory Nat Rev Immunol 16, 90-101.
Safinia, in view of Yu and Hippen fails to teach T reg cell populations of CD45RAhi CD25hi CD127lo CD4+ Tregs of claim 56 and 57.
Rosenblum, however, teaches T reg populations of this subset are considered resting T cells (Table 2) and further highlights these cells have not yet been exposed to antigens but can be to develop T reg memory cells which will further expand upon restimulation with the same antigen (pg.91, col 1, line 10 - pg.91, col 2, line 16 / Figure 1).
Thus, Safinia in view of Yu and Hippen discloses a method of expanding T reg cells comprising at least a 1st stimulation step, a 1st rest step and a 2nd stimulation step for about 1-5 days each, and Rosenblum teaches the resting T reg cell population of the claimed phenotype and further that they represent a population of resting T reg cells which can be conditioned to further expand and perform better upon restimulation with a specific antigen. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to try the combination of the teachings of Safinia in view of Yu and Hippen informed by the teachings of Rosenbum to select a resting T reg cell population, with a reasonable expectation of success, to develop of a method of expanding Treg cells wherein the T reg cells are selected for their ability to have a more robust response to restimulation for improved expansion.
Response to Arguments
Applicant's arguments filed 18 June 2026 have been fully considered but they are not persuasive.
Applicant’s arguments with respect to claims 1-3, 7, 10, and 48-49 have been considered but are moot because the new 103 rejection necessitated by the amendment of the claims to add time constraints to the independent claim.
In response to applicant's argument that “the combination of Safinia and McDonald cannot render the present claims obvious” (pg.10, lines 9-10). The argument is against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). Safina teaches discontinuous stimulation while MacDonald teaches various methods of stimulation, combining the reference supports the obviousness to try discontinuous stimulation as taught in Safinia with one of the methods in McDonald which have different benefits.
In response to applicant's argument that “the combination of Safinia Yu and Hippen fails to provide the timing required by the presently claims methods” (pg.11, line 22 – pg.12, line 2). The argument is against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). Safina teaches discontinuous stimulation while Yu teaches the improved efficiency of genetically engineering stimulated T cell populations and further demonstrates this stimulation can take place over 1-3 days (a shorter period than tried in Safinia), Finally Hippen further supports Safinia providing more support to trying a discontinuous stimulation method by teaching restimulation expanded T reg expansion. Combining the references supports obviousness.
Applicant’s arguments with respect to claim 29 has been considered but is moot because of the new U.SC. 103 rejection to independent claim 1 necessitated by the amendment of the claims to add time constraints to the independent claim.
Applicant’s arguments with respect to claims 29, 31, 42, 33, and 44 “that the combination of references does not teach the time constraint of the instant claims” have been considered but is moot because of the new U.SC. 103 rejection to independent claim 1 necessitated by the amendment of the claims to add time constraints to the independent claim.
In response to applicant’s arguments that “the combination of references fails to provide any reason to expect the dramatically different improved results” of the instant claims. In the MPEP 2143(I)(E) it is stated “that the claim would have been obvious if that “a person of ordinary skill has a good reason to pursue the known options within his or her technical grasp. If this leads to the anticipated success, it is likely that product [was] not of innovation but of ordinary skill and common sense”, further in the MPEP it is stated that “obviousness does not require absolute predictability” (MPEP 2143.02(II)) and also states “conclusive proof of efficacy is not required to show a reasonable expectation of success” (MPEP 2143.02(I)). Thus, the combined references give known options within their technical grasp which have been seen to improve T regulatory cell expansion, which would lead to obviousness to try and thus the anticipated success.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/D.A.A/Examiner, Art Unit 1647 /JOANNE HAMA/Supervisory Patent Examiner, Art Unit 1647
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