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 .
Claim Status
1. Claims 14-17,19-20,24-25,39-45,47-55,57-67,69-83,85-87 and 89-93 remain pending and are under consideration. Claims 84 and 88 have been canceled.
Election/Restrictions
2. Applicant’s election without traverse of Group II (claims 14 – 17, 19 – 21, 24, and 25) in the reply filed on 03/26/2024 is acknowledged.
3. Claims 18, 22, 23, and 26 -38 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 03/26/2024.
Priority
4. The present application is a continuation of PCT/US2019/051865, filed September 19, 2019, which claims priority to U.S. Patent Application Serial No. 62/734,019 filed September 20, 2018.
Information Disclosure Statement
5. The information disclosure statement (IDS) submitted on 11/20/2025 is acknowledged. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Withdrawn Claim Rejections
6. The rejection of claims 84 and 88 under 35 U.S.C. 103 is rendered moot in view of Applicant’s cancellation of these claims.
Claim Interpretation
7. Claims 14, 40, 50, and 62 recite “2-deoxyglucose (2DG) in an amount sufficient to selectively deplete thymocytes, relative to stromal cells” but do not recite a mass, volume, concentration, or quantity of 2DG or even a range of masses, volumes, concentrations, or quantities. For the purpose of applying prior art, “in an amount sufficient to selectively deplete thymocytes, relative to stromal cells” will be interpreted as treating thymic tissue with 2DG such that more thymocytes are reduced than stromal cells.
Maintained Claim Rejections
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
8. Claims 14 – 17, 19 – 20, 24, 25, 39 – 45, 47 – 55, 57, 59 – 67, 69, 71 – 83, 85 – 87, and 89 remain rejected under 35 U.S.C. 103 as being unpatentable over Maharlooei (Maharlooei, MK et. al. (2017), IXA 2017- Abstracts of the 14th Congress of the International Xenotransplantation Association, Baltimore, USA. Xenotransplantation, 24: 15 – 16, September 11, 2017; previously cited), hereinafter Maharlooei which is cited in the IDS filed 03/26/2024, in view of Stefanelli (STEFANELLI, Claudio, et al. Biochemical Journal 322.3 (1997): 909-917; previously cited), hereinafter Stefanelli in view of Fudaba (Fudaba, Yasuhiro, et al. The Journal of Immunology 181.11 (2008): 7649-7659; previously cited), hereinafter Fudaba which is cited on the IDS filed 03/17/2021in view of Parent (US 20160010055 A1; previously cited), hereinafter Parent, which is cited on the IDS filed 03/17/2021. Although maintained, note that the rejection is revised in light of the cancellation of claims 84 and 88.
Maharlooei teaches a method for making a human/pig hybrid thymus comprising treating fetal swine thymic tissue with 2-deoxyglucose (2DG) (claim 14(i), 16, 19, 40(i), 42, 44, 50(i), 52, 54, 62(i), 64, 66) and injecting human thymic epithelial cells (TECs) isolated from human thymi (“second mammalian species is a human” and “second mammalian species is a primate” of claim 14(a), 20, 40(a), 45, 50(a), 55, 62(a), 67) suspended in Matrigel into the fetal swine thymic tissue treated with 2DG (claim 14(ii), 15, 24, 25, 40(ii), 41, 47, 48, 50(ii), 51, 59, 60, 62(ii), 63, 71, 72) (page 16, Methods).
Regarding “in an amount sufficient to selectively deplete thymocytes, relative to stromal cells” of claims 14(i), 40(i), 50(i), and 62(i), Maharlooei teaches thymic stromal cells were isolated, resuspended in Matrigel and injected into freeze-thawed fetal SW thymic tissue that had been treated with 2-DG (page 15 panel B and C and Figure legend; page 16, left col. Methods). Therefore, Maharlooei teaches the amount of 2-DG used to reduce thymocytes did not affect stromal cells as Maharlooei concludes that injection of human thymic stromal cells into pig thymus is an effective approach to generate human/pig hybrid thymus because HuTEC-injected thymi were functional and supported human thymopoiesis in humanized mice and the panels B and C show white arrows that represent human TECs inside the pig thymus (page 15 figure panels B and C and figure legend; page 16, left col. Results and Conclusion).
Regarding claims 74, 78, 82, and 86, Maharlooei teaches a human/pig hybrid thymus to overcome the challenges in xenotransplantation in humans that are T cells selected on swine MHC in pig thymus may not optimally recognize antigens presented by human MHC and swine TECs do not display human tissue-restricted antigens (TRAs) that might impair negative selection and also a lack of Tregs specific for human TRAs (page 15, right col. para. 1; page 16, left col. para. 1).
Maharlooei does not teach a numerical amount of 2-DG used or length of treatment “of claims 14(i), 40(i), 50(i), 62(i), 77, 81, 85, or 89 or generating thymic epithelial cells from human iPSCs derived from the first mammalian species of claims 14(b), 40(b) or embryonic stem cells of claims 50(b), 62(b) or “miniature swine” of claims 17, 43, 53, and 65 or 100 mM 2DG for 12 hours of claims 39, 49, 61, and 73 or iPSCs are from the subject of claims 75 and 79 or embryonic stem cells are from the subject of claims 83 and 86 or embryonic stem cells share HLA alleles with the second mammalian species of claims 57 and 69 or the iPSCs share HLA alleles with the second mammalian species of claims 76 and 80. However, Maharlooei teaches treating thymic tissue with 2DG suppresses glycolysis to reduce the numbers of pig thymocytes in the fetal SW thymus (page 16, left col. paragraph 2). Maharlooei teaches powerful immune responses to xenografts are difficult to control with conventional immunosuppression without excessive toxicity (page 15, right col. para. 1). Maharlooei teaches thymus transplantation is a promising approach to induce T cell tolerance for xenotransplantation and humanized mice generated with human hematopoietic stem cells and swine thymus grafts are tolerant to both species (page 1, right col. paragraph 1). Maharlooei teaches T cells selected on swine MHC in pig thymus may not optimally recognize antigens presented by human MHC and swine TECs do not display human tissue-restricted antigens (page 15, right col. para. 1; page 16, left col. para. 1). Maharlooei teaches that a human/pig hybrid thymus might overcome challenges associated with inducing T cell tolerance for xenotransplantation (page 2, left col. paragraph 1). Maharlooei teaches human/pig hybrid thymi were functional and supported human thymopoiesis in humanized mice and contained the injected human TECs (page 16, Results).
Regarding a numerical amount of 2-DG to selectively deplete thymocytes relative to stromal cells and length of treatment of claims 14(i), 40(i), 50(i), 62(i), 77, 81, 85, and 89, Stefanelli teaches treating thymocytes with 20 mM 2-DG for 24 hours resulted in cell death as determined by lactate dehydrogenase leakage into the medium from damaged cells (page 910, left col. para. 1 and right col. para. 5; Table 2; page 912, left col. para. 2). Stefanelli teaches inhibition of mitochondrial respiration with oligomycin or sodium cyanide did not affect cell viability but inclusion of 2-deoxyglucose increased cell death and ATP levels fell to less than 10% of control (page 912, left col. para. 2). Stefanelli does not teach “miniature swine” of claims 17, 43, 53, and 65 or 100 mM 2DG for 12 hours of claims 39, 49, 61, and 73 or generating thymic epithelial cells from human iPSCs derived from the first mammalian species of claims 14(b), 40(b) or embryonic stem cells of claims 50(b), 62(b) or iPSCs are from the subject of claims 75 and 79 or embryonic stem cells are from the subject of claims 83 and 86 or embryonic stem cells share HLA alleles with the second mammalian species of claims 57, 69, 84, and 88 or the iPSCs share HLA alleles with the second mammalian species of claims 76 and 80.
Regarding claims 39, 49, 61, and 73, Fudaba teaches a hybrid thymic tissue comprising porcine thymus and mouse thymic epithelial cells (mTECs) were grafted into nude mice (page 7652, left col. para. 2; page 7656, left col.; Figure 8). Fudaba teaches that without mTECs, the nude mice developed autoimmunity (page 7656, left col.). Fudaba teaches autoimmunity could not be suppressed by thymocytes from nude mice with porcine xenografts (FGP mice) and that these thymocytes caused autoimmunity (page 7655, right col. para. 3). Fudaba teaches this demonstrates that selection in a porcine thymic xenograft results in an intrinsic tendency of T cells to cause autoimmune disease (page 7655, right col. para. 3). Fudaba teaches transplantation allows excellent peripheral reconstitution of mouse Tregs (page 7652, left col. para. 2; Figure 1A). Fudaba teaches the inability of thymocytes from FPG nude mice to suppress the autoimmunity induced by T cells from FPG nude mice most likely reflects a reduced ability of the porcine thymic epithelium to positively select Treg that recognize murine MHC-peptide complexes (page 7657, left col. para. 5). Fudaba teaches the thymic epithelium plays an important role in the positive selection of Treg (page 7657, left col. para. 5). Fudaba teaches consistently, the engraftment of normal mouse mTECs in porcine thymus grafts reduced the capacity of thymocytes from porcine grafts to induce autoimmunity (page 7657, right col. para. 3). Fudaba teaches purified host-type thymic epithelial cells can ameliorate disease, persist long-term within the thymic xenograft and allow the development of regulatory cells that can inhibit autoimmunity upon adoptive transfer (page 7658, left col. para. 1). One would have been motivated to combine the teachings of Maharlooei regarding a method for making a hybrid thymic tissue for xenotransplantation with the teachings of Stefanelli regarding killing of thymocytes by 2-DG with the teachings of Fudaba regarding hybrid porcine/mouse thymic xenografts that prevent autoimmunity in a method for making a hybrid thymic tissue for xenotransplantation where thymocytes are treated in an amount sufficient to deplete pig thymocytes to reduce autoimmunity but not deplete pig stromal cells to allow peripheral reconstitution of mouse Tregs because Maharlooei teaches powerful immune responses to xenografts are difficult to control with conventional immunosuppression without excessive toxicity and thymus transplantation is a promising approach to induce T cell tolerance for xenotransplantation and Fudaba teaches thymocytes from FGP mice caused autoimmunity and Fudaba teaches selection in a porcine thymic xenograft results in an intrinsic tendency of T cells to cause autoimmune disease. Further, it would be obvious to adjust the 2DG treatment taught by Maharlooei and Stefanelli and length of treatment taught by Stefanelli since it is a result effective variable dependent on the number of pig thymocytes depleted to minimize autoimmunity while maintaining pig stromal cells for the development of Tregs.
Regarding “miniature swine” of claims 17, 43, 53, and 65, Fudaba teaches a miniature swine fetal pig thymus/liver fragment where miniature swine fetuses were used as donors of porcine thymic tissue (page 7650, left col. para. 1 – 2).
Fudaba does not teach thymic epithelial cells generated from iPSCs of claims 14(b), and 40(b) or thymic epithelial cells generated from embryonic stem cells of claims 50(b) and 62(b) or iPSCs are from the subject of claims 75 and 79 or embryonic stem cells are from the subject of claims 83 and 86 or embryonic stem cells share HLA alleles with the second mammalian species of claims 57, 69, 84, and 88 or the iPSCs share HLA alleles with the second mammalian species of claims 76 and 80. However, Fudaba teaches fetal porcine thymic grafts can efficiently support human thymopoiesis resulting in the generation of a T cell repertoire that is specifically tolerant of the xenogeneic donor (Abstract; page 7656, right col. para. 2). Fudaba teaches using vascularized porcine thymic grafts in a nonhuman primate model has allowed acceptance of life-supporting α-1,3-galatosyl transferase knockout porcine kidney grafts for months with no evidence of rejection (page 7656, right col. para. 2). Fudaba teaches human T cells developing in porcine thymic grafts repopulate the periphery (page 7658, left col. para. 2).
Regarding thymic epithelial cells generated from iPSCs of claims 14(b), 40(b), 75 and 79 and the iPSCs share HLA alleles with the second mammalian species of claims 76 and 80, Parent teaches a method of generating thymic epithelial cells from human iPSCs (claim 75, 76, 79, 80) that are functional when transplanted in vivo (page 1, 0006; page 5, 0064 and 0070 – 0074; page 7, 0110, and Figure 1A). Parent teaches the pluripotent stem cells used in the method may be an established stem cell line (page 7, 0110). Parent teaches stem cell-derived TECs support T cell development in vivo (page 18, 0261 and Figure 3). Parent teaches TECs generated from patient-specific iPS cell lines may also be used as a tool to model human disease (page 14, 0220).
Regarding thymic epithelial cells generated from embryonic stem cells of claims 50(b), 62(b), 83, and 87 and embryonic stem cells share HLA alleles with the second mammalian species of claims 57, 69, 84, and 88, Parent teaches a method of generating thymic epithelial cells from human embryonic stem cells (claims 57, 69, 83, 84, 87, 88) that are functional when transplanted in vivo (page 1, 0006, 0018; page 5, 0070 – 0074; page 7, 0110, and Figure 1A). Parent teaches the stem cells used in the method may be an established embryonic stem cell line (page 7, 0110). Parent teaches stem cell-derived TECs support T cell development in vivo (page 18, 0261 and Figure 3).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Maharlooei regarding a method for making a hybrid thymic tissue for xenotransplantation with the teachings of Stefanelli regarding killing of thymocytes by 2-DG with the teachings of Fudaba regarding hybrid porcine/mouse thymic xenografts that prevent autoimmunity with the teachings of Parent regarding a method of generating thymic epithelial cells from human iPSCs or human embryonic stem cells to arrive at the claimed methods for making a hybrid thymic tissue where thymic tissue from a pig is treated with 2DG in an amount sufficient to selectively deplete thymocytes relative to stromal cells and thymic epithelial cells from a human are introduced into the 2DG-treated thymic tissue. One would be motivated to combine the teachings of Maharlooei, Stefanelli, Fudaba, and Parent in a method in a method for making a hybrid pig/human thymic tissue for xenotransplantation to reduce autoimmunity as Maharlooei teaches that a human/pig hybrid thymus might overcome challenges associated with inducing T cell tolerance for xenotransplantation and Fudaba teaches selection in a porcine thymic xenograft results in an intrinsic tendency of T cells to cause autoimmune disease. One would have a reasonable expectation of success in combining the teachings as Maharlooei teaches human/pig hybrid thymus was functional and supported human thymopoiesis in humanized mice and contained the injected human TECs and Fudaba teaches fetal porcine thymic grafts can efficiently support human thymopoiesis resulting in the generation of a T cell repertoire that is specifically tolerant of the xenogeneic donor and using vascularized porcine thymic grafts in a nonhuman primate model has allowed acceptance of life-supporting α-1,3-galatosyl transferase knockout porcine kidney grafts for months with no evidence of rejection.
9. Claims 58 and 70 remain rejected under 35 U.S.C. 103 as being unpatentable over Maharlooei (Maharlooei, MK et. al. (2017), IXA 2017- Abstracts of the 14th Congress of the International Xenotransplantation Association, Baltimore, USA. Xenotransplantation, 24: 15 – 16, September 11, 2017; previously cited), hereinafter Maharlooei which is cited in the IDS filed 03/26/2024, in view of Stefanelli (STEFANELLI, Claudio, et al. Biochemical Journal 322.3 (1997): 909-917; previously cited), hereinafter Stefanelli in view of Fudaba (Fudaba, Yasuhiro, et al. The Journal of Immunology 181.11 (2008): 7649-7659; previously cited), hereinafter Fudaba which is cited on the IDS filed 03/17/2021in view of Parent (US 20160010055 A1; previously cited), hereinafter Parent, which is cited on the IDS filed 03/17/2021. as applied to claims 14 – 17, 19 – 20, 24, 25, 39 – 45, 47 – 55, 57, 59 – 67, 69, 71 – 83, 85 – 87, and 89 in further view of Riolobos (Riolobos L, et. al. Mol Ther. 2013 Jun;21(6):1232-41; previously cited), hereinafter Riolobos, in view of Sachs (US 20060147428 A1; previously cited), hereinafter Sachs.
Maharlooei in view of Stefanelli, Fuduba, and Parent make obvious the limitations of claims 50 and 62 as set forth above. Maharlooei, Stefanelli, Fuduba, and Parent do not teach genetically engineered embryonic stem cells to share HLA alleles with the second mammalian species of claims 57 and 70. However, Maharlooei teaches the second mammalian species is human in human/pig hybrid thymus that was functional and supported human thymopoiesis in humanized mice and contained the injected human TECs that were HLA-DR+ (page 15 Figure and page 16, Results).
Riolobos teaches transplanted cells and organs can be rejected based on their HLA type and typically prolonged treatment with immunosuppressive drugs is required to prevent rejection of mismatched grafts, often with dangerous side effects (page 1232). Riolobos teaches one solution to prevent rejection is to use autologous stem cells derived from each patient (page 1232).
Sachs teaches a donor thymic graft in which host T cells can mature where the thymic tissue is swine and the host is a primate, e.g., a human (page 1, 0004 – 0006). Sachs teaches genetically engineering cells by inserting a nucleic acid which expresses human MHC genes, i.e., the HLA genes (page 2, 0015 and page 17, 0227). Sachs teaches genetically engineering cells by inserting donor MHC genes (i.e., swine) would promote acceptance of a graft by the recipient (i.e., human) (page 2, 0015).
It would have been obvious prior to the effective filing date of the invention as claimed for the person or ordinary skill in the art to substitute the human thymic epithelial cells (TECs) isolated from human thymus as taught by Maharlooei with TECs generated by the method of Parent using embryonic stem cells as taught by Riolobos that are genetically engineered to share HLA alleles as taught by Sachs to arrive at a method for making a hybrid thymic tissue. One would be motivated to make the substitution to alleviate the need for immunosuppressive drugs and prevent cell/organ rejection because Riolobos teaches the use of immunosuppressive drugs can have dangerous side effects and because Sachs teaches genetically engineering by inserting a nucleic acid which expresses human HLA genes results in tolerance to thymic tissue from the donor. One would have a reasonable expectation of success in carrying out the substitution because Parent teaches the stem cell-derived TECs support T cell development in vivo.
10. Claims 90 – 93 are rejected under 35 U.S.C. 103 as being unpatentable over Maharlooei (Maharlooei, MK et. al. (2017), IXA 2017- Abstracts of the 14th Congress of the International Xenotransplantation Association, Baltimore, USA. Xenotransplantation, 24: 15 – 16, September 11, 2017; previously cited), hereinafter Maharlooei which is cited in the IDS filed 03/26/2024, in view of Stefanelli (STEFANELLI, Claudio, et al. Biochemical Journal 322.3 (1997): 909-917; previously cited), hereinafter Stefanelli in view of Fudaba (Fudaba, Yasuhiro, et al. The Journal of Immunology 181.11 (2008): 7649-7659; previously cited), hereinafter Fudaba which is cited on the IDS filed 03/17/2021in view of Parent (US 20160010055 A1; previously cited), hereinafter Parent, which is cited on the IDS filed 03/17/2021. as applied to claims 14 – 17, 19 – 20, 24, 25, 39 – 45, 47 – 55, 57, 59 – 67, 69, 71 – 83, 85 – 87, and 89 in further view of Cao (Cao, Yilin, et. al. PloS one 9.8 (2014): e104104; previously cited), hereinafter Cao.
Maharlooei in view of Stefanelli, Fuduba, and Parent make obvious the limitations of claims 50, 40, 50, and 62 as set forth above. Maharlooei teaches treating thymic tissue with 2DG (page 16, left col., Methods) but does not teach CD4/CD8 cell changes as a result of 2DG. Fudaba teaches CD4 T cells generated in porcine thymus xenografts have an increased tendency to induce autoimmunity (page 7655, left col. last para.). Fudaba teaches CD4+CD8- cells accelerated autoimmunity and transfer of thymocytes from FPG nude mice alone also caused autoimmunity (page 7655, right col. last para.). Fudaba teaches in Figure 2 that the higher dose of CD8+ cells does not suppress autoimmune disease (Figure 2).
Regarding (a) – (c) of claims 90 – 93, Cao teaches culturing CD4 and CD8 cells with 2-deoxyglucose where the viability of CD4 and CD8 cells was similarly affected by increasing inhibitor dose (page 8, left col. para. 1; Figure 9B). Cao teaches CD4 growth was significantly more sensitive to 2-DG than CD8 cells at lower concentrations and CD8 cell growth may be more resistant to 2-DG (page 6, right col. last para.; page 8, left col. para. 1; Figure 9B; page 10, right col. para. 1).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Maharlooei regarding a method for making a hybrid thymic tissue for xenotransplantation with the teachings of Stefanelli regarding killing of thymocytes by 2-DG with the teachings of Fudaba regarding hybrid porcine/mouse thymic xenografts that prevent autoimmunity with the teachings of Parent regarding a method of generating thymic epithelial cells from human iPSCs or human embryonic stem cells with the teachings of Cao regarding 10 mM 2-DG reduces CD4 and CD8 cell viability to arrive at the claimed methods for making a hybrid thymic tissue where the thymic tissue treated with 2DG has reduced ratios of double positive CD4 and CD8 cells to double negative cells. One would be motivated to combine the teachings of Maharlooei, Stefanelli, Fudaba, Parent, and Cao in a method in a method for making a hybrid pig/human thymic tissue for xenotransplantation to reduce autoimmunity as Maharlooei teaches that a human/pig hybrid thymus might overcome challenges associated with inducing T cell tolerance for xenotransplantation and Fudaba teaches CD4+ and CD8+ cells do not suppress autoimmunity. One would have a reasonable expectation of success in combining the teachings as Maharlooei teaches human/pig hybrid thymus was functional and supported human thymopoiesis in humanized mice and contained the injected human TECs and Fudaba teaches fetal porcine thymic grafts can efficiently support human thymopoiesis resulting in the generation of a T cell repertoire that is specifically tolerant of the xenogeneic donor and using vascularized porcine thymic grafts in a nonhuman primate model has allowed acceptance of life-supporting α-1,3-galatosyl transferase knockout porcine kidney grafts for months with no evidence of rejection and Cao teaches 10 mM 2-DG reduces CD4 and CD8 cell viability.
Applicant’s Arguments/ Response to Arguments
11. Applicant Argues: On page 13, paragraph 3 – 4, page 14, para. 1 – 2, page 15, para. 2, page 17, para. 2, Applicant asserts that none of the cited references teach or suggest treating thymic tissue with 2-DG in an amount sufficient to selectively deplete thymocytes relative to stromal cells. Applicant asserts on page 14, para. 1 that Stefanelli merely discloses experiments involving isolated thymocyte suspensions that lack any stromal elements.
Response to Arguments: Maharlooei teaches injecting stromal cells into fetal swine thymic tissue treated with 2-DG to suppress glycolysis to reduce the numbers of pig thymocytes in the fetal swine thymus (page 16, left col. Methods). As Maharlooei teaches the stromal cells survive when combined with the 2-DG treated thymocytes, Maharlooei teaches selective depletion of thymocytes relative to stromal cells with 2-DG. Independent claims 14, 40, 50, and 62 do not recite a numerical value or unit (concentration, mass, volume) of 2DG sufficient to selectively deplete thymocytes relative to stromal cells. Should applicant amend the claims to recite a numerical value or range of values, the rejection over Maharlooei may be overcome.
As previously cited, Stefanelli teaches treating thymocytes with 20 mM 2-DG for 24 hours resulted in cell death as determined by lactate dehydrogenase leakage into the medium from damaged cells (page 910, left col. para. 1 and right col. para. 5; Table 2; page 912, left col. para. 2). Stefanelli teaches inhibition of mitochondrial respiration with oligomycin or sodium cyanide did not affect cell viability but inclusion of 2-deoxyglucose increased cell death and ATP levels fell to less than 10% of control (page 912, left col. para. 2). One of ordinary skill in the art would recognize that Stefanelli results teach that thymocytes are sensitive to 2-DG, which inhibits glycolysis, but not by inhibitors of oxidative phosphorylation (oligomycin, sodium cyanide). Therefore, one of ordinary skill in the art would be motivated to combine the teachings of Maharlooei and Stefanelli and have a reasonable expectation of success in combining their teachings in a method of making a hybrid thymic tissue suitable for xenotransplantation because Maharlooei teaches treating fetal swine thymic tissue with 2DG and injecting human TECs isolated from human thymi suspended in Matrigel into the fetal swine thymic tissue treated with 2DG, where treating thymic tissue with 2DG suppresses glycolysis to reduce the numbers of pig thymocytes in the fetal SW thymus (page 16, Methods; page 16, left col. para. 2) and Maharlooei teaches human/pig hybrid thymi were functional and supported human thymopoiesis in humanized mice and contained the injected human TECs (page 16, Results) and the human/pig hybrid thymus might overcome challenges associated with inducing T cell tolerance for xenotransplantation (page 2, left col. paragraph 1) and Stefanelli teaches an amount of 2DG that resulted in cell death of thymocytes.
Conclusion
No claims allowed.
THIS ACTION IS MADE FINAL. 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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/Z.M.B./Examiner, Art Unit 1632
/MARCIA S NOBLE/Primary Examiner, Art Unit 1632