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
This action is in response to the papers filed August 13, 2026.
Claim Amendments
Applicant’s amendment to the claims filed 08/13/2026 is acknowledged.
Claims 3, 8-14 have been cancelled.
Claims 1-2, 4, 6-7 are amended.
Claims 1-2, 4-7 are pending and under examination.
Election/Restrictions
The following is a summary of the restriction/election requirements in the application. See, the Requirement for Restriction/Election mailed 03/30/2026. In the reply filed 05/18/2026, Applicant elected without traverse Invention I, drawn to a method of making an animal model of a CD19 CAR-T cell therapy for leukemia complicated by CRS.
Priority
The instant application 18/439,283 was filed on 02/12/2024. This application claims priority based on Chinese patent application CN202310751294.0 filed 06/25/2023. Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55. While a certified copy of the foreign patent application is provided with the instant application, a certified English translation of said foreign patent application has not been provided.
Withdrawal of Prior Rejections/Objections
Rejections and/or objections not reiterated from the previous Office action mailed 06/02/2026 are hereby withdrawn. The following rejections and/or objections are either newly applied or are reiterated and are the only rejections and/or objections presently applied to the instant application. Applicant’s remarks filed 08/13/2026 have been carefully considered but are not moot in view of the new grounds of rejection set forth in this action.
Claim Objections
Claim 1 is objected to because of the following informalities:
In claim 1, line 3, the phrase “comprising following steps” should be “comprising the following steps” instead.
Appropriate action is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-2, 4-7 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
This rejection is newly applied, necessitated by amendment.
Step (g) of claim 1 recites “administering the amplified CD19 CAR-T cells at different number into the leukemia tumor-bearing animal.” Generally, it is unclear what the recitation “at different number” is intended to convey or how the recitation further limits step (g). Indeed, the number of CAR-T cells administered is defined by the later recitation “wherein the number of the administered CD19 CAR-T cells is 107-108,” so it is unclear how the earlier recitation “administering the amplified CD19 CAR-T cells at different number” modifies the administration step. For these reasons, one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. Dependent claims are included in the basis of the rejection because they do not correct the deficiencies of the claim upon which they depend.
Claim 7 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
This rejection is newly applied, necessitated by amendment.
Claim 7 recites “[t]he preparation method of an animal model of a CD19 CAR-T cell therapy for leukemia complicated by CRS according to claim 1, preparing CD19 CAR-T cell populations comprising following steps ...” The recitation is generally unclear as drafted. For example, there is no clear nexus between the recitation “preparing CD19 CAR-T cell populations” and an earlier limitation, and, therefore, it is unclear if claim 7 is intended to define new steps or further limit steps (a) through (e) of claim 1. It appears that the recitation should have been “[t]he preparation method of an animal model of a CD19 CAR-T cell therapy for leukemia complicated by CRS according to claim 1, wherein preparing CD19 CAR-T cell populations comprises the following steps ...” in order to provide a logical structure to the claim. For these reasons, one of ordinary skill in the art would not be reasonably apprised of the scope of the invention.
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 1-2, 4-5 and 7 are rejected under 35 U.S.C. 103 as being unpatentable over Arcangeli et al. (2022) “CAR T cell manufacturing from naive/stem memory T lymphocytes enhances antitumor responses while curtailing cytokine release syndrome” The Journal of clinical investigation, 132(12), 15 pages.
Arcangeli discloses an animal model of CD19 CAR-T cell therapy for leukemia complicated by CRS comprising an immunodeficient mouse having a reconstituted human immune system (HuSGM3) and engrafted with the B-cell acute lymphoblastic leukemia cell line Nalm-6. See, e.g., Abstract, pages 1-3, and Figures 2A and 4A.
Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy and patient donors, and CD3 T cells were isolated from PBMCs by FACS®. T cells were subsequently stimulated (activated) with Miltenyi® MACS-GMP T-Cell TransAct™, transduced with a lentiviral vector (LV) encoding a CD19 CAR, and amplified in Miltenyi® TexMACS™ culture media supplemented with IL-7/IL-15 for 15 days. See, page 12.
Immunodeficient mice having a reconstituted human immune system (HuSGM3) were infused with 0.5×106 Nalm-6 cells expressing Lucia® luciferase and, 5 or 7 days after, treated with a 1×106 or 1×107 CD19 CAR T cells. See, pages 12-13.
The difference between the instantly claimed invention and that of Arcangeli is that the T cells are sorted from the PBMCs by “magnetic bead sorting” (claim 1). Arcangeli sorts cells using FACS®, i.e., fluorescence-activated cell sorting. However, prior to the effective filing date of the instantly claimed invention, magnetic-activated cell sorting, or MACS®, was a well-known and common alterative to FACS® for sorting T cells from PBMCs. Official Notice taken, if necessary. Therefore, it would have been prima facie obvious to one of ordinary skill in the art to substitute FACS® sorting, as found in Arcangeli, with magnetic bead sorting or MACS®, as known in the prior art, with a reasonable expectation of success because magnetic bead sorting or MACS® is a well-known and common alterative means for sorting T cells from PBMCs, and the simple substitution of one known element for another would have yielded predictable results to one of ordinary skill in the art at the time of the invention.
For these reasons, the method according to claim 1 would have been prima facie obvious over the prior art.
The dependent claims further recite the number of CD19 CAR-T cells in the CD19 CAR-T cell line is 50-200 times the number of Nalm-6 cells in the Nalm-6 cell line (claim 2); the Nalm-6 cell line has 105, 2 × 105, 5 × 105 or 106 Nalm-cells (claim 4); the number of CD19 CAR-T cells in the CD19 CAR-T cell line is 100 times the number of Nalm-6 cells in the Nalm-6 cell line (claim 5); and the CAR T cells are amplified until the number of CAR T cells is greater than 108 (claim 7).
Arcangeli discloses that immunodeficient mice having a reconstituted human immune system (HuSGM3) were infused with 0.5×106 Nalm-6 cells expressing Lucia® luciferase and, 5 or 7 days after, treated with a 1×106 or 1×107 CD19 CAR T cells. See, pages 12-13.
Generally, differences in amount or concentration will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such amount or concentration is critical. "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). In this case, the difference between the animal model instantly claimed and that taught by the prior art lies in the number or amount of CD19 CAR-T cells and Nalm-6 cells. Accordingly, absent a secondary consideration, said differences in number or amount of CD19 CAR-T cells and Nalm-6 cells would have been prima facie obvious over the prior art. See, MPEP 2144.05.
Regarding dependent claim 7, the claim further recites that the CD3 T cells are activated in a culture medium supplemented with IL2. Arcangeli discloses stimulating (activating) T cells with Miltenyi® MACS-GMP T-Cell TransAct™. See, page 12. Arcangeli does not disclose supplementing the medium with IL-2. However, prior to the effective filing date of the instantly claimed invention, IL-2 was a well-known media supplement for T cell activation and expansion. Official Notice taken, if necessary. Therefore, it would have been prima facie obvious to one of ordinary skill in the art to modify the culture media of Arcangeli to further comprise IL-2, as known in the prior art, with a reasonable expectation of success because IL-2 supplementation supports T cell activation and expansion.
For these reasons, and those outlined above, dependent claims 2, 4-5 and 7 would have also been prima facie obvious over the prior art.
Claims 6 is rejected under 35 U.S.C. 103 as being unpatentable over Arcangeli et al. (2022) “CAR T cell manufacturing from naive/stem memory T lymphocytes enhances antitumor responses while curtailing cytokine release syndrome” The Journal of clinical investigation, 132(12), 15 pages, as applied above; in further view of Pegram et al. (2015) “IL-12-secreting CD19-targeted cord blood-derived T cells for the immunotherapy of B-cell acute lymphoblastic leukemia” Leukemia, 29(2), 415-422.
Arcangeli does not disclose the immunodeficient mouse is a SCID/Beige mouse or that the Nalm-6 cells are labelled with GFP and luciferase (GFP-luc), as claimed in dependent claim 6.
Pegram is relevant prior art for disclosing the adoptive transfer of CD19 CAR T cells into tumor-bearing SCID-Beige mice. Mononuclear cells were separated from human umbilical cord blood (UCB); T cells were isolated, activated and expanded with CD3/CD28 magnetic beads in a medium supplemented with IL-2; and T cells were retrovirally transduced with a CD19 CAR. SCID-Beige mice were administered 1×106 Nalm-6 tumor cells expressing a GFP-firefly luciferase fusion protein (Nalm6-eGFP-FFluc), followed by administration of 5×106 CAR T cells. See, e.g., Abstract and page 416.
Therefore, prior to the effective filing date of the instantly claimed invention, it would have been prima facie obvious to one of ordinary skill in the art to modify the invention of Arcangeli by using a SCID/Beige mouse and Nalm-6 cells labeled with GFP-luc with a reasonable expectation of success because the SCID/Beige mouse was a well-known immunodeficient mouse model used in adoptive transfer studies, and the GFP-luc dual reporting system enables both bioluminescence and fluorescence imaging for monitoring tumor challenges.
Conclusion
The following prior art made of record and not relied upon is considered pertinent to applicant's disclosure:
Bove et al. (02 Jan 2023) “CD4 CAR-T cells targeting CD19 play a key role in exacerbating cytokine release syndrome, while maintaining long-term responses” Journal for immunotherapy of cancer, 11:e005878, 13 pages, newly cited.
Bove discloses an animal model of CD19 CAR-T cell therapy for leukemia complicated by CRS comprising an immunodeficient mouse having a reconstituted human immune system (HuSGM3) and engrafted with the B-cell acute lymphoblastic leukemia cell line Nalm-6. See, e.g., Abstract and Figures 3A, 5A and 6A.
Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of a donor, and CD3 T cells were isolated from PBMCs using Miltenyi® Pan T-Cell Isolation Kit (i.e., magnetic bead sorting). T cells were subsequently separated into CD4 and CD8 fractions, stimulated (activated) with Miltenyi® MACS-GMP T-Cell TransAct™, and transduced with a lentiviral vector (LV) encoding a CD19 CAR. See, page 2, right column.
Immunodeficient mice having a reconstituted human immune system (HuSGM3) were infused with 0.5×106 Nalm-6 cells expressing luciferase (Lucia®, InvivoGen) and, 5 or 7 days after, treated with a low dose or high dose of CD19 CAR T cells. See, page 3 and Figures 3, 5-6.
Shan et al. (2021) “Establishment of a cytokine release syndrome associated with chimeric antigen receptor T cell treatment in SCID/Beige mice model” Zhonghua Zhong liu za zhi [Chinese Journal of Oncology], 43(12), 1248-1254, of record. A paper copy of original document, which includes an English-language Abstract, and a human-assisted machine translation of the full document, generated by the Scientific & Technical Information Center (STIC) at the USPTO, have been provided in a previous Office action.
Shan discloses a CRS mouse model related to CAR-T cell therapy. CAR-T cells targeting human CD19 molecule were constructed by molecular cloning and lentiviral transfection. The CD19 CAR-T cells were injected into tumor-bearing SCID/Beige mice, and the CRS phenomenon was verified, thereby providing an animal model for the mechanism of CAR-T treatment-related CRS and CRS prevention strategies. See, English-language Abstract.
The CAR-T cells were prepared by collecting peripheral blood from healthy individuals, collecting peripheral blood mononuclear cells using density gradient centrifugation, and selecting CD3+ T cells through magnetic bead sorting. The CD3+ T cells were then activated by stimulation with CD3 and CD28 immunomagnetic beads, and the selected CD3+ T cells were infected by a lentivirus expressing the CD19 CAR. See, pages 5-6 and 8, and Figure 3, of the translation.
The tumor-bearing mice were prepared by injecting a high-dose or a low-dose of Raji cells into SCID/Beige mice, and CAR-T cells were injected seven days after tumor formation. Raji is a human lymphoma cell line. See, pages 4 and 6 of the translation. Shan also performed a CAR-T cell functional assay by co-incubating CD19 CAR-T cells with Raji or Nalm-6 cells, both of which are CD19-postiive tumor cell lines, at different effector-target ratios. CD19 CAR-T cells were similarly effective in targeting both Raji and Nalm-6 cells relative to control CD19-negative K562 tumor cells. Shan further recognizes that the Nalm-6 cell line is a human acute lymphoblastic leukemia cell line. See, pages 4-5 and 9, and Figure 5, of the translation.
Giavridis et al. (2018) “CAR T cell–induced cytokine release syndrome is mediated by macrophages and abated by IL-1 blockade” Nature medicine, 24(6), 731-738, newly cited.
Giavridis discloses a murine model of CRS following CAR-T cell infusion. Primary human T cells were purified from buffy coats of healthy donors by negative magnetic selection and transduced with a CD19 CAR. For CRS model, SCID-Beige mice were administered Raji GFP-firefly luciferase (FLuc) cells and, afterwards, administered CAR T cells. In a stress-test model, immunodeficient mice (NSG) were administered Nalm-6-GFP-FLuc cells and, afterwards, administered CAR T cells. See, e.g., Abstract and first page of Methods.
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.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JAMES J GRABER whose telephone number is (571)270-3988. The examiner can normally be reached Monday-Thursday: 9:00 am - 4:00 pm.
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/JAMES JOSEPH GRABER/Examiner, Art Unit 1631
