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 .
DETAILED ACTION
The text of those sections of Title 35, U.S. Code not included in this action can be found
in a prior Office action.
This Action is in response to the papers filed on December 19, 2025 for a Request for Continued Examination. A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on December 19, 2025 has been entered.
Pursuant to the amendment filed on December 19, 2025, claims 69, 74-76 and 95, and 99 are currently pending. Claims 69 and 74 have been amended, and claim 96 has been cancelled, and claim 99 is newly filed in Applicant' s amendment filed on December 19, 2025.
The restriction requirement between Groups I-IV was previously made FINAL in the Office Action dated April 18, 2025.
Therefore, claims 69, 74-76 and 95, and 99 are currently under examination to which the following grounds of rejection are applicable.
Priority
The present application is a 35 U.S.C. 371 national stage filing of the International Application No. PCT/US2020/044033, filed July 29, 2020. Applicant’s claim for the benefit of a prior-filed parent provisional application 62/932,342 filed on November 7, 2019 and 62/881,311 filed on July 31, 2019 under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, or 365(c) is acknowledged.
Thus, the earliest possible priority for the instant application is July 31, 2019.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on December 19, 2025 was filed. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Response to Arguments
Withdrawn Objections/Rejections in response to Applicants’ arguments or amendments:
Claim Rejections - 35 USC § 103
In view of Applicant’s amendments filed on December 19, 2025, wherein claims 69 and 74 have been amended, and claim 96 has been cancelled, and claim 99 is newly filed, the rejection to claims 69, 74-76 and 95 rejected under 35 U.S.C. 103 as being unpatentable over Baek et al. (Anticancer Research 33.5 (2013): 2011-2019) in view of Wu et al. (Frontiers in immunology 8 (2017):930), Kerbauy et al. (US Pub No. 2020/0390816 A1), and Kamiya et al. (US 11,896,616 B2), have been withdrawn. The rejection to now cancelled claim 96 has been rendered moot. The withdrawn rejection is in view of the amendment to claim 69 and Applicants’ arguments regarding activation/stimulation steps, and the instant claims being directed to no such step.
Applicants' arguments are moot in view of the withdrawn rejection. A response to any argument pertaining to a new or maintained rejection can be found below.
New Grounds of Rejection:
Claim Rejections - 35 USC § 103
Claims 69, 74-76, 96, and 99 are newly rejected under 35 U.S.C. 103 as being unpatentable over Baek et al. (Anticancer Research 33.5 (2013): 2011-2019; of record) in view of Wu et al. (Frontiers in immunology 8 (2017):930), Kerbauy et al. (US Pub No. 2020/0390816 A1; of record IDS filed on February 12, 2025), and Romee et al. (Blood, The Journal of the American Society of Hematology 120.24 (2012): 4751-4760).
Claim 69 is directed to a method for expanding natural killer cells, comprising: (a) culturing a population of NK cells with a feeder cell population in a culture media, having soluble interleukin 2 (IL2) at a concentration of less than 50 IU/mL, soluble interleukin 12 (IL12) at a concentration of between about 0.01 ng/mL and about 8 ng/mL and soluble interleukin 18 (IL18) at a concentration of between about 0.01 ng/mL and about 30 ng/mL, wherein the feeder cell population comprises cells, which express 4-1BB ligand ( 4-1BBL) and membrane-bound interleukin-15 (mbIL15); and (b) contacting the NK cells with a vector encoding a chimeric antigen receptor (CAR).”
Regarding claim 69, Baek teaches a method for enhancing the expansion of natural killer cells, comprising: culturing a population of natural killer (NK) cells with a feeder cell population in a culture media, wherein the feeder cell population comprises cells engineered to express 4-1BBL and membrane-bound interleukin-15 (mbIL15); and wherein the culture media comprises interleukin 2 (IL2) (“We compared NK cell expansion using cryopreserved-irradiated feeder cells (cryopreserved group) and freshly-irradiated feeder cells (fresh group). To expand NK cells, peripheral blood mononuclear cells were isolated and co-cultured with-100 Gy-irradiated K562 leukemia cells that had been modified to express 4-1BB ligand and membrane-bound (mb) interleukin (IL)-15 (K562-mb15-41BBL cells) for three weeks in the presence of IL-2 and IL-15.” (abstract); “Every other day, the medium was replaced with fresh medium containing 10 IU/ml human IL-2.”(p 2, col 2, par 1)).
Baek does not teach culturing a population of NK cells with a feeder cell population in a culture media, having soluble interleukin 12 (IL12) at a concentration of between about 0.01 ng/mL and about 8 ng/mL and soluble interleukin 18 (IL18) at a concentration of between about 0.01 ng/mL and about 30 ng/mL, and contacting the NK cells with a vector encoding a chimeric antigen receptor (CAR).
Kerbauy teaches using IL-12 and IL-18 for the activation of NK cells prior to culturing with feeder cells that express 4-1BBL and membrane-bound interleukin-15 (mbIL15); “there is provided an in vitro method for expanding NK cells comprising obtaining a population of NK cells; pre-activating the population of NK cells in a pre-activation culture comprising an effective concentration of IL-12, IL-15, and IL-18 to obtain pre-activated NK cells; and expanding the pre-activated NK cells in an expansion culture comprising artificial antigen presenting cells (aAPCs) expressing CD137 ligand, thereby producing expanded NK cells.” (par 5; CD137 ligand is 4-1BBL); “In certain aspects, the aAPCs further express a membrane-bound cytokine. In some aspects, the membrane-bound cytokine is membrane-bound IL-21 (mIL-21) or membrane-bound IL-15 (mIL-15)… In certain aspects, the leukemia-cell derived aAPCs are further defined as K562 cells engineered to express CD137 ligand and/or mIL-21.” (par 7). In reference to the produced NK cells, “The present studies showed that the pre-activated and expanded NK cells produced by the present methods exhibited enhanced anti-tumor functionality against leukemia and lymphoma cell lines. Pre-activated and expanded NK cells also displayed enhanced antibody-dependent cellular cytotoxicity (ADCC).” (par 35).
Kerbauy teaches the method further comprising contacting the population of NK cells with a vector encoding a chimeric antigen receptor (CAR), wherein the wherein the CAR is configured to target one or more of CD19, a ligand of the natural killer receptor group D (NKG2D), CD70, or BCMA (“In further aspects, the NK cells are engineered to express a chimeric antigen receptor (CAR). In some aspects, the CAR comprises a CD19, .., or BCMA antigen-binding domain” (par 12)).
Kerbauy teaches culturing a population of NK cells with a feeder cell population in a culture media, having soluble interleukin 12 (IL12) at a concentration of between about 0.01 ng/mL and about 8 ng/mL and soluble interleukin 18 (IL18) at a concentration of between about 0.01 ng/mL and about 30 ng/mL (“In certain aspects, the pre-activation culture comprises IL-18 and/or IL-15 at a concentration of 10-100 ng/mL, such as 40-60 ng/mL, particularly about 50 ng/mL. In some aspects, the pre-activation culture comprises IL-12 at a concentration of 0.1-150 ng/mL, such as 1-20 ng/mL, particularly about 10 ng/mL.” (par 0008); (par 0008); “In particular embodiments, the pre-activation cytokines may be IL-12, IL-15, and IL-18. One or more additional cytokines may be used for the pre-activation step. The pre-activation may be for a short period of time such as 5-72 hours, such as 10-50 hours, particularly 10-20 hours, such as 12, 13, 14, 15, 16, 17, 18, 19, or 20 hours, specifically about 16 hours.”(par 0055).
Lastly, Kerbauy teaches the use of recombinant versions of IL-2, IL-12, and IL-18 (rhIL-2 , rhIL-12, and rhIL-18), all of which are expected to be soluble in solution (par 0057). Moreover, it remains obvious to use a soluble version of the cytokines since they are added to an aqueous culture media wherein the co-culturing occurs.
Likewise, Wu teaches the response to NK cells to different cytokines, e.g. IL-2, IL-12 and IL-18. IL-2 is considered “an immune-stimulatory cytokine … [that] has been shown that activated T cell-derived IL-2 can enhance NK cell responses toward infection in vivo and can activate NK cells in vitro… IL-2 may have an important role in maintaining NK cell activity…it is needed for NK cell effector functions but is not indispensable for their development” (p 5, col 1). In reference to the cytokines IL-12 and IL-18, “previous reports have shown that preactivation with IL-12, IL-15, and IL-18 induces CD25 expression on NK cells, which suggests that pretreatment of NK cells with the above cytokines can enhance NK cell responsiveness to IL-2 and may lead to better IL-2 treatment efficacy.”; “Prior exposure to IL-15 sensitizes NK cells to secondary stimuli, referred to as “priming,” thereby resulting in exaggerated responses. Previous studies have shown that IL-12 can induce elevated IFN-γ production in IL-15-primed NK cells.” (p 5 col 2). Additionally, it is also described that IL-18 is important for promoting the production of IFN-γ by NK cells against viral, fungal, bacterial, and parasitic infections. The review further states, “Recently, researchers also found that the combination of these two cytokines [IL-12 and IL-18] can reverse NK cell anergy and increase the survival rate of mice bearing MHC-deficient tumors. These findings imply that IL-18 can play stronger roles when combined with other stimulatory cytokines; therefore, combining cytokines together is better for achieving enhanced efficacy.” (p 7, col 1). Lastly, Figure 2 depicts modified K562 feeder cells applied for the expansion of primary NK cells using IL-2; and furthermore IL-12, 15 and 18 are used for preactivation and/or mbIL-15 can be expressed for improving NK cell survival or antitumor function.
PNG
media_image1.png
544
809
media_image1.png
Greyscale
Lastly, in reference to the limitation pertaining to the inclusion of the claimed cytokines with feeder cells, Romee teaches NK cells cocultured with feeder cells, i.e. K562 cells, and describes “For preactivation with K562 leukemia cells, freshly purified NK cells were cocultured with irradiated (3000 rads) K562 cells for 16 hours at a 4:1 ratio either in presence of rhIL-15 (1 ng/mL) alone or in combination with rhIL-12 (10 ng/mL), rhIL-18 (50 ng/mL), or rhIL-12 (10 ng/mL) + rhIL-18 (50 ng/mL).” (p 4752, col 1, par 2). The outcomes were “human NK cells preactivated briefly with cytokine combinations including IL-12, IL-15, and IL-18 followed by a 7- to 21-day rest have enhanced IFN-γ production after restimulation with IL-12 + IL-15, IL-… Therefore, human NK cells have functional memory-like properties after cytokine activation, which provides a novel rationale for integrating preactivation with combinations of IL-12, IL-15, and IL-18 into NK cell immunotherapy strategies.” (abstract).
It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have been motivated in using IL-12 and IL-18 during the culturing of NK cells and feeder cells that express mbIL15 and 4-1BBL in culture media comprising IL-2 based on Kerbauy and Wu teaching the stimulating/activation activity of these cytokines on NK cell expansion, and moreover in view of Romee teaching this claimed combination with feeder cells leads to retention of a memory-like phenotype, and enhanced IFN-γ production by NK cells. Therefore, there is a reasonable expectation that by including these cytokines with the claimed feeder cells and NK cells, there would likely lead to successfully enhancement of the expansion of NK cells. This is based on Wu teaching the role/outcomes of using these cytokines, Kerbauy teaching the increased activation of NK cells by culturing with IL-12 and IL-18, and Romee teaching enhanced IFN-γ production with the introduction of cytokines while feeder cells are present.
Additionally, it would be obvious to optimize the concentrations of the claimed cytokines, e.g. IL-12 and IL-18, and the timing of culturing with select cytokines as a way to improve the methods taught by the prior art of expanding NK cells.
In reference to combined stimulation and expansion with cytokines for NK cells and feeder cells, the Romee reference teaches this step. Regardless, it remains obvious to have stimulation prior to or during expansion based on Kerbauy teaching improved cytotoxicity obtained from stimulated NK cells, and more specifically NK stimulated prior to expansion rather than after (Fig 2A-C, 8-9). Kerbauy further teaches both these steps are important for enhanced cytotoxicity as seen in Fig. 6, wherein only stimulation or expansion did not lead to the same outcomes as the combination. Altogether, it would be obvious to try in combining these steps as it was known that stimulated NK cells are important for cytoxicity, and that stimulation after expansion results in a reduced cytoxicity in comparison to before NK expansion, and based on a finite amount of options the combination of steps would an obvious choice.
Regarding claim 74, dependent on claim 69, Baek and Kerbauy teach wherein the culture media is further supplemented for a second time with IL2 at a concentration that is greater than that present in the culture media in (a) (Baek: “Every other day, the medium was replaced with fresh medium containing 10 IU/ml human IL-2. After one week, the concentration of IL-2 was increased to 100 IU/ml,” (p 2012, col 2); Kerbauy : “In additional aspects, the expansion culture further comprises IL-2. In some aspects, the IL-2 is present at a concentration of 10-500 U/mL, such as 100-300 U/mL, particularly about 200 U/mL… In some aspects, the IL-2 is replenished in the expansion culture every 2-3 days.” (par 011)).
Regarding claim 75, dependent on claim 69, Baek teaches wherein the feeder cell population comprises K562 cells, wherein the K562 cells are irradiated prior to co-culture, and wherein the K562 cells express both 4-1BBL and mbIL15 (“We compared NK cell expansion using cryopreserved-irradiated feeder cells (cryopreserved group) and freshly-irradiated feeder cells (fresh group). To expand NK cells, peripheral blood mononuclear cells were isolated and co-cultured with-100 Gy-irradiated K562 leukemia cells that had been modified to express 4-1BB ligand and membrane-bound (mb) interleukin (IL)-15 (K562-mb15- 41BBL cells)” (abstract)).
Regarding claim 76, dependent on claim 69, the rejection above to claim 69 describes that Kerbauy teaches the method further comprising contacting the population of NK cells with a vector encoding a chimeric antigen receptor (CAR), wherein the wherein the CAR is configured to target one or more of CD19, a ligand of the natural killer receptor group D (NKG2D), CD70, or BCMA (“In further aspects, the NK cells are engineered to express a chimeric antigen receptor (CAR). In some aspects, the CAR comprises a CD19, .., or BCMA antigen-binding domain” (par 12)).
Regarding claim 95, dependent on claim 74, Baek teaches wherein the concentration of the IL2 in the culture media after the further supplementation is less than 500 IU/mL (“Every other day, the medium was replaced with fresh medium containing 10 IU/ml human IL-2. After one week, the concentration of IL-2 was increased to 100 IU/ml,” (p 2012, col 2))
Regarding claim 99, dependent on claim 69, Baek and Kerbauy teach wherein the soluble interleukin 12 (IL12) and soluble interleukin 18 (IL18) in the culture media increases the IFN-y production by NK cells.
In reference to the “wherein” clause, MPEP 2111.04 states regarding wherein and whereby clauses: a “whereby clause in a method claim is not given weight when it simply expresses the intended result of a process step positively recited.’" Id. (quoting Minton v. Nat’l Ass’n of Securities Dealers, Inc., 336 F.3d 1373, 1381, 67 USPQ2d 1614, 1620 (Fed. Cir. 2003)).” In this case, the wherein clause, “wherein the soluble interleukin 12 (IL12) and soluble interleukin 18 (IL18) in the culture media increases the IFN-y production by NK cells”, expresses the desired result of the positive step of including IL12 and IL18 in the culture. Thus, the clause is not given weight and does not limit the claims.
Regardless, Kerbauy teaches IFN-y production by NK cells is increased after activation with cytokines and subsequent expansion, as seen in Figures 1A-F, par 0024. Furthermore, as described in the claim 69 rejection above, Wu teaches that IL12 and IL18 promote IFN-y production of NK cells, along with Romee teaching enhanced IFN-γ production after culturing with IL-12 and IL-18 (abstract).
Response to Applicants’ Arguments as they apply to rejection of claims 69, 74-76, and 95 under 35 USC § 103
Starting on page 4 of the remarks filed on December 19, 2025, Applicants essentially argue the following:
In relation to claim 69, Applicants’ state the cited references do not teach or suggest the claimed subject matter, and “In other words, the method recited in claim 69 does not require a separate pre-activation or culturing of the NK cells with a combination of interleukins prior to culturing with feeder cells as disclosed in Kerbauy.”
Applicant states, “Furthermore, the data presented in Kerbauy demonstrates that NK cells that were preactivated and expanded exhibited superior cytotoxicity as compared to NK cells that were expanded but not pre-activated. See figures 2A-2C. Furthermore, Kerbauy also discloses that the sequence or order at which NK cells were pre-activated and expanded was important. Specifically, NK cells that were pre-activated prior to expansion resulted in superior expansion and highly cytotoxic NK cells as compared to expansion followed by pre-activation. See figures 8-9.”
Applicant states in regard to Kerbauy teaching away from the claimed invention, “Furthermore, pre-activated NK cells in Kerbauy were stimulated with irradiated K562-based feeder cells expressing membrane-bound IL-21 and CD137-ligand. See Kerbauy at [0105]. It was well known to a person of skill in the art at the time of invention that NK cells expanded with mbIL-21 have improved proliferation of human NK cells with longer telomeres and less senescence compared to NK cells expanded with mbIL-15 feeder cells. (see Denman et al. - submitted herewith in an IDS). Therefore, a person of skill in the art with the knowledge of” the state of art at the time of invention and upon reviewing Kerbauy would be motivated to use feeder cells expressing mbIL-21 and would be dissuaded from using mbIL-15 in view of the prior art. Accordingly, the claims are non-obvious over the cited references. “
In response to these arguments they have been fully considered but are not persuasive due to the following reasons:
Regarding the first presented argument, the new rejection is based on the combination of Baek et al., Wu et al., Kerbauy et al. which were used in the prior Office Action for rejecting claims 69, 74-76, and 95, and now in view of Romee for the rejection of these claims, and newly filed claim 99.
In response to applicant's arguments 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). The rejections above make clear that Baek in view of Kerbauy, Wu, and Romee teach the entirety of the method of claim 69, and therefore outcomes related to NK expansion and/or cytotoxicity are to be expected. Kerbauy teaches pre-activated and expanded NK cells transduced to express a chimeric antigen receptor (CAR) against CD19, were able to target and destroy cells more efficiently than controls (Fig. 4A). Furthermore, Romee teaches the stimulation of feeder cells and NK with the claimed IL-12 and IL-18 leading to enhanced production of IFNγ (abstract, Methods). Moreover, the rejection makes it clear that the outcomes of using the claimed cytokines are well-known in the art, and that there remains a reasonable expectation that including the stimulation step with expansion would lead to similar outcomes as have been shown in the art with just stimulation using the cytokines followed by expansion.
Regarding the second argument, in particular Kerbauy’s Fig 2A-C, these figures merely describe that the use of cytokines, e.g. IL-2, IL-18 and IL-15, improved cytotoxicity of NK cells in comparison to cultured NK in the absence of cytokines (par 0025). Therefore, this suggests that use of these cytokines boosts NK cell cytoxicity properties. Additionally, Figures 8 and 9 depict improved outcomes of activation followed by expansion as opposed to the inverse order; wherein there is an improved cytotoxicity. The instant claim still stimulates with cytokines, and is done during culturing which is different than what is depicted in Fig 8. It is noted that Fig. 6 shows that both stimulation and expansion resulting in superior cytotoxicity in comparison to CB-NK cells that were either pre-activated but not expanded, or expanded but not pre-activated. Therefore, it remains obvious to include stimulation and the expansion of NK cells, and that the decision of combining these steps is merely an option from a finite list of options, as there would a reasonable expectation that similar outcomes of enhanced cytotoxicity would be observed from the obtained NK cells.
Regarding the third argument, Kerbauy clearly teaches the membrane-bound cytokine is membrane-bound IL-21 (mIL-21) or membrane-bound IL-15 (mIL-15) (par 0007), and moreover Baek teaches “Recently, remarkable rates of highly cytotoxic NK cell expansion were reported using a protocol that employs irradiated, genetically-engineered K562 cells, such as K562-based artificial antigen-presenting cells with membrane-bound IL-21 (mbIL21) and the K562 cell line modified to express membrane-bound IL-15 (mbIL15) and 4-1BB ligand (K562-mb15-41BBL)-used in the present study (7-9).” (p 2011-2012). Therefore, it can be understood that the art teaches various feeder cells, particularly genetically-engineered K562 cells, that can be used in the expansion of NK cells with high cytotoxicity.
As states in MPEP 2143.01(1). “The disclosure of desirable alternatives does not necessarily negate a suggestion for modifying the prior art to arrive at the claimed invention…The court stated that "the prior art’s mere disclosure of more than one alternative does not constitute a teaching away from any of these alternatives because such disclosure does not criticize, discredit, or otherwise discourage the solution claimed…””. In the instant case, Kerbauy’s use of feeder cells expressing IL-21 does not teach away from feeder cells expressing mbIL-15.
Provisional Double Patenting
Claim 69, 74-76 and 95 remain provisionally rejected and claim 99 is newly provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1 of copending Application No. 18/024,012 as per claims filed on November 14, 2025 (hereinafter ‘012) in view of Kamiya et al. (US 11,896,616 B2; citations taken from U.S. Pub No. 2020/0016208 A1; of record IDS filed on February 12, 2025).
This is a provisional nonstatutory double patenting rejection.
Claim 1 of ‘012 is directed to “A method for enhancing the expansion of natural killer (NK) cells, comprising: (a) co-culturing, in a culture media, a population of NK cells with a first population of feeder cells for a first period of time, wherein the first feeder cell population comprises cells engineered to express 4-1 BBL and membrane-bound interleukin-15 (mbIL15), wherein the population of NK cells comprises fewer cells than the first population of feeder cells, wherein the culture media comprises interleukin 2 (IL2), and one of interleukin 12 (IL12) and interleukin 18 (IL18), and wherein the co-culturing for the first period of time results in an expanded population of NK cells; (b) separating, after the first period of time, at least a portion of the expanded population of NK cells from the feeder cells, (c) co-culturing, in fresh culture media, the at least a portion of the expanded population of NK cells with a second population of the feeder cells for a second period of time, wherein the at least a portion of the expanded population of NK cells comprises fewer cells than the second population of feeder cells, wherein the culture media comprises interleukin 2 (IL2), and one of interleukin 12 (IL12) and interleukin 18 (IL18), and wherein the co-culturing for the second period of time results in a further expanded population of NK cells.”
Claim 3 of ‘012 is directed to “The method of Claim 1, wherein: (i) the IL2 is present in the media at a concentration between about 10 units/mL and about 100 units/mL (ii), if the IL12 is present in the media, the IL12 is at a concentration between about 10 ng/mL and about 100 ng/mL: and (iii), if the IL18 is present in the media, the IL18 is at a concentration between about 0.01 ng/mL and about 30 ng/mL, wherein each of the first and the second periods of time is about 7 days, and wherein the co-culturing is repeated at least three times.
Claim 9 of ‘012 is directed to “The method of Claim 1, further comprising contacting the NK cells with a vector encoding a chimeric antigen receptor (CAR).”
Claim 69 of the instant application is directed to: “A method for expanding natural killer (NK) cells, comprising:
(a) culturing a population of NK cells with a feeder cell population in a culture media having soluble interleukin 2 (IL2) at a concentration of less than 50 IU/mL, soluble interleukin 12 (IL12) at a concentration of between about 0.01 ng/mL and about 8 ng/mL and soluble interleukin 18 (IL18) at a concentration of between about 0.01 ng/mL and about 30 ng/mL, wherein the feeder cell population comprises cells, which express 4-1BB ligand ( 4-IBBL) and membrane-bound interleukin-15 (mbIL15);
(b) contacting the NK cells after (a) with a vector encoding a chimeric antigen receptor (CAR).
The claims are similar in that they are directed to a method for enhancing the expansion of natural killer cells wherein they both require culturing a population of natural killer (NK) cells with a feeder cell population in a culture media, wherein the feeder cell population comprises cells engineered to express 4-1BBL and membrane-bound interleukin-15 (mbIL15); and wherein the culture media comprises interleukin 2 (IL2), and one of soluble interleukin 12 (IL12) and soluble interleukin 18 (IL18). Moreover, the claims are similar in being directed to similar ranges pertaining to the amount of cytokines to be used in the culture media, and moreover contacting the NK cells with a vector encoding a chimeric antigen receptor (CAR).
The claims are different in that ‘012 requires the expansion of NK be intended for immunotherapy, the first period of time for co-culturing using a population of NK cells that comprises fewer cells than the population of feeder cells, a second co-culture/ period of time involving the same elements set forth for the first period of time, and an optional step of repeating the separating and co-culturing steps. Lastly, ‘012 has amended the claims wherein the culture media contains one of IL12 and IL18 as opposed to both for which the instant application is directed to.
In reference to the recitation of the expanded NK cells being used in immunotherapy this merely considered an intended usage as the claim does not specifically recite the active step of using these cells in such therapy and therefore does not hold weight.
In reference to the limitation of using fewer NK cells than feeder cells and a second period of time, these limitations are made obvious over Kamiya et al. that teaches using a higher ratio of feeders to NK cells, and furthermore the feeders are freshly added after every two weeks (“To renew the potential expansion of the NK cells, fresh genetically-modified K562 cells were added every 2 weeks, at a K562:NK ratio of 5:1, and IL-2 concentration was maintained at 40 IU/mL during the first week and at 400 IU/mL subsequently, after T cell depletion.” (Example 2)). Moreover, it would be quite obvious to have a higher ratio of the feeder cells that are intended to support the growth of cells in culture, e.g. NK cells, by secreting essential growth factors and providing a substrate. Furthermore, feeder cells are mitotically inactivated to prevent cell division, and would therefore require to be freshly added to maintain growth factor supplementation since cell growth is impacted. This extends to having a second period of time for co-culturing because feeder cells would need to be freshly added, and moreover having a second co-culture would an obvious step in view of further expanding NK cells for downstream applications.
For these reasons, it would have been prima facie obvious for ‘012’ to have claimed a second period of time in view of further expanding NK cells and for new reagents (e.g. feeder cells, fresh media) being required after a period of co-culturing time as this is routine in the art.
In reference to the culture media containing IL12 and IL18 for which the instant application is directed to, and ‘012 being directed to culture media with one of IL12 and IL18, it can be seen that the scopes overlap wherein ‘012 method allows for both IL12 and IL18 specifically one being used in step (a) and the other being used in step (c); wherein the interleukins are rather separated within the culturing process.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Response to Applicants’ Arguments as they apply to rejection of claims 69, 74-76, 95, and 99 under provisional double patenting:
At page 9 of the remarks filed on December 19, 2025, applicants essentially argue “ Additionally, the Manual of Patent Examining and Procedure (MPEP) states, "If a provisional nonstatutory double patenting rejection is the only rejection remaining in an application having the earlier patent term filing date, the examiner should withdraw the rejection in the application having the earlier patent term filing date and permit that application to issue as a patent, thereby converting the provisional nonstatutory double patenting rejection in the other application into a nonstatutory double patenting rejection upon issuance of the patent." See Manual of Patent Examining and Procedure § 804.1.B. l .(b )(i). Accordingly, withdrawal of the rejection is respectfully requested.’ Such is not persuasive. Note that because the claims are not allowable as written, the provisional double patenting rejection of record stands per MPEP 1504.06.
Conclusion
Claims 69, 74-76 and 95, and 99 are rejected. No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to MICHAEL A RIGA whose telephone number is (571)270-0984. The examiner can normally be reached Monday-Friday (8AM-6PM).
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Maria G Leavitt can be reached at (571) 272-1085. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/MICHAEL ANGELO RIGA/ Examiner, Art Unit 1634
/TERESA E KNIGHT/ Primary Examiner, Art Unit 1634