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 .
Status of the Claims
Claims 1, 4-5 and 9-12 are pending.
Claims 10-12 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim and made FINAL. Election was made without traverse in the reply filed on 07/31/2023.
Claims 1, 4-5, and 9 have been examined on their merits.
Withdrawn Objections & Rejections
The objections and rejections presented herein represent the full set of objections and rejections currently pending in the application. Any objections or rejections not specifically reiterated are hereby withdrawn.
The rejection of claims 1 and 9 under 35 U.S.C. 103 as being unpatentable over Liddelow et al. (Nature, 2017, on IDS 01/15/2021, previously cited, hereafter “Liddelow I”) in view of Williams et al. (PLoS One, 2009, on IDS 01/15/2021, previously cited) and Rivieccio et al. (The Journal of Immunology, 2005, previously cited) as evidenced by Foo et al. (Neuron, 2011, previously cited) is withdrawn in order to incorporate Rubio et al. (Immunology, 2011).
The rejection of claim 4 under 35 U.S.C. 103 as being unpatentable over Liddelow et al. (Nature, 2017, on IDS 01/15/2021, previously cited, hereafter “Liddelow I”) in view of Williams et al. (PLoS One, 2009, on IDS 01/15/2021, previously cited) and Rivieccio et al. (The Journal of Immunology, 2005, previously cited), as evidenced by Foo et al. (Neuron, 2011, previously cited), as applied to claim 1 above, and further in view of Embad et al. (Stem cells and Development, 2012, previously cited) is withdrawn in order to incorporate Rubio et al. (Immunology, 2011).
The rejection of claim 5 under 35 U.S.C. 103 as being unpatentable over Liddelow et al. (Nature, 2017, on IDS 01/15/2021, previously cited, hereafter “Liddelow I”) in view of Williams et al. (PLoS One, 2009, on IDS 01/15/2021, previously cited) and Rivieccio et al. (The Journal of Immunology, 2005, previously cited), as evidenced by Foo et al. (Neuron, 2011, previously cited), as applied to claim 1 above, and further in view of Liddelow et al. (Immunity Review, 2017, previously cited, hereafter “Liddelow II”) is withdrawn in order to incorporate Rubio et al. (Immunology, 2011).
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.
Claims 1 and 9 are rejected under 35 U.S.C. 103 as being unpatentable over Liddelow et al. (Nature, 2017, on IDS 01/15/2021, previously cited, hereafter “Liddelow I”) in view of Rubio et al. (Immunology, 2011), Williams et al. (PLoS One, 2009, on IDS 01/15/2021, previously cited) and Rivieccio et al. (The Journal of Immunology, 2005, previously cited) as evidenced by Foo et al. (Neuron, 2011, previously cited).
In regards to claim 1, in regards to step (a), Liddelow I teaches a method for generating A1 astrocytes (Abstract, p481; Methods, p488).
It is noted that an “A1” astrocyte is a termed coined by Liddelow I to distinguish between polarized subsets of astrocytes (Introduction, p481; i.e., A1 and A2 astrocytes). Specifically, A1 astrocytes are destructive to neural synapses while A2 astrocytes are protective (Introduction, p481).
Methodologically, Liddelow I teaches that purified astrocytes derived from forebrain (thus, primary astrocytes) are cultured in media comprising TNFα for 24 hours (Abstract, p481; Methods, p488). A timing of 24 hours overlaps with the range of 1 day or more (see MPEP 2144.04). Liddelow I teaches that the culturing is performed in serum-free conditions (Methods, Immunopanning and cell culture, p488; p482).
In regards to the forebrain astrocytes, Liddelow I teaches that the method for obtaining the astrocytes derives from Foo (Methods, p488). As evidenced by Foo, astrocytes obtained by this method strongly promote synapse formation (Summary, p799). As further taught by Liddelow I, A1 astrocytes are specifically destructive to synapses, and form specifically in response to neuroinflammation (p481). Moreover, Liddelow I demonstrates that the primary astrocytes (before treatment with TNFα) retained their non-activated in vivo gene profile and did not exhibit an A1-astrocyte phenotype with a variety of other factors tested (see Extended Data, Fig. 2a & b).
Therefore, because the forebrain astrocytes, as taught by Liddelow I are explicitly known to promote synapse formation, that A1 astrocytes are destructive to synapses and form as a result of neuroinflammation, and do no exhibit an A1-astrocyte phenotype prior to treatment with TNFα, a person of ordinary skill in the art would have recognized that the forebrain astrocytes, from which the A1 astrocytes, as taught by Liddelow I are generated, are not themselves A1 astrocytes.
In regards to the concentration, Liddelow I teaches that primary astrocytes were cultured with 30 ng/mL TNFα (Abstract, p481; Methods, p488).
While a concentration of 30 ng/mL TNFα is greater than the claimed range of 0.5 to 10 ng/mL, a person of ordinary skill in the art could have arrived at a concentration of 0.5 to 10 ng/mL by routine optimization and the disclosure does not point to a criticality in this concentration range (see MPEP 2144.05(II)(A), generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature 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) (Claimed process which was performed at a temperature between 40°C and 80°C and an acid concentration between 25% and 70% was held to be prima facie obvious over a reference process which differed from the claims only in that the reference process was performed at a temperature of 100°C and an acid concentration of 10%.); see also Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382 ("The normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine where in a disclosed set of percentage ranges is the optimum combination of percentages."); In re Hoeschele, 406 F.2d 1403, 160 USPQ 809 (CCPA 1969) (Claimed elastomeric polyurethanes which fell within the broad scope of the references were held to be unpatentable thereover because, among other reasons, there was no evidence of the criticality of the claimed ranges of molecular weight or molar proportions).
In the instant case, because Rubio teaches that astrocytes can be treated with 10 ng/mL TNFα in vitro for the production of pathological cytokines (Fig. 5, p366), a person of ordinary skill in the art could have arrived at the claimed concentration of 0.5 to 10 ng/mL by routine optimization with predicable results and a reasonable expectation of success.
In regards to IFNγ, while Liddelow I teaches that published microarray datasets indicates that IFNγ promotes expression of A1-specific transcripts (markers), Liddelow I does not explicitly teach that A1 astrocytes were generated with IFNγ.
However, a person of ordinary skill in the art would have been motivated to modify the method of Liddelow I and add IFNγ because Williams teaches that IFNγ and TNFα work together to promote CXCL10 expression in specifically a pro-inflammatory context (Abstract, p1; Figure 1, p3; p8), and as taught by Liddelow I, A1 astrocytes express CXCL10 (Figure 1, p482).
They would have been further motivated to add IFNγ because Williams also teaches that pro-inflammatory cytokines IFNγ and TNFα are elevated in the brains of patients with HIV-associated neurocognitive disorders (HAND,) such neuroinflammatory conditions such as HIV-encephalitis (HIVE) (Introduction, p1, p8), and a person of ordinary skill in the art would have been motivated to mimic the in vivo milieu of these patients in order to more accurately study human disease.
Moreover, they would have been motivated to add IFNγ because as taught by Rivieccio it promotes IL-1β-induced iNOS (thus, pro-inflammatory) expression (p3724).
Furthermore, because as above Liddelow I teaches that IFNγ promotes expression of A1 markers, because Williams demonstrates that the combination of IFNγ and TNFα promotes astrocyte CXCL10 expression in a pro-inflammatory context, and because Rivieccio teaches that astrocytes can be contacted with IFNγ in a pro-inflammatory context, it could have been done with predictable results and a reasonable expectation of success.
In regards to the concentration of IFNγ, Williams teaches that IFNγ was used at a concentration of 50 ng/mL. While, greater than the range of 0.1 ng/mL to 20 ng/mL, a person of ordinary skill in the art could have arrived at a concentration range of 0.5 ng/mL to 20 ng/mL by routine optimization, and the disclosure does not point to a criticality in this amount (see MPEP 2144.05(II)(A) as discussed above).
In the instant case, because Rivieccio teaches that primary human astrocyte cultures can be treated with 10 ng/mL IFNγ (Results, p3721), which lies in the range of 0.1 to 20 ng/mL, a person of ordinary skill in the arts could have arrived at this concentration by routine optimization with predictable results and a reasonable expectation of success.
Moreover, a person of ordinary skill in the arts would have been motivated to reduce the amount of IFNγ used as to minimize the amount of reagents needed, which would save time and expenses. Furthermore, because Rivieccio teaches that 10 ng/mL IFNγ was effective for culturing primary human astrocyte and induced CXCL10 expression (Results, p3721), it could have been done with predictable results and a reasonable expectation of success.
Additionally, Liddlow I teaches that the forebrain astrocytes are derived from rats or mice, not humans specifically.
However, Liddelow I also teaches that A1 astrocytes are abundant in various human neuroinflammatory and neurodegenerative diseases including Alzheimer’s, Huntington’s and Parkinson’s disease, amyotrophic lateral sclerosis and multiple sclerosis (Abstract, p481; A1 astrocytes in human disease), and promotes cell death in human cells (Figure 4, p485). Therefore, a person of ordinary skill in the art would have been motivated to select human cells because it would have been most relevant for studying human health and disease.
Furthermore, because Rivieccio teaches that primary human astrocytes can be used for the testing of cytokines (Title, Abstract, p3719), it could have been done with predictable results and a reasonable expectation of success.
In regards to step (1), Liddelow I teaches that forebrain astrocytes were cultured in serum-free conditions for 6 days (Immunopanning and cell culture, p488), which overlaps with the range of at least 24 hours.
In regards to step (2), Liddelow I teaches that following this, astrocytes were treated with TNFα for 24 hours (Immunopanning and cell culture, p488), which also overlaps with the timing of at least 1 day after the step (1). It would have been predictably obvious to supplement the medium with IFNγ to induce expression of CXCL10 as discussed above.
In regards to the properties recited in claim 1, “wherein . . . the human A1 astrocytes have neuronal cytotoxicity”, “wherein the neuronal cytotoxicity shortens the length of neurites”, and “wherein the human A1 astrocytes express CXCL10”, these are all known inherent properties of A1 astrocytes themselves.
As taught by Liddelow I, A1 astrocytes are neurotoxic and kill neurons (and thus are cytotoxic) (p481); shorten neurite outgrowth (Extended Data Figure 5k); and express CXCL10 (Figure 1, p482).
In regards to claim 9, as optimized, a concentration of 10 ng/mL TNFα and 10 ng/mL IFNγ results in a 1:1 ratio which overlaps with the claimed range.
Therefore, the combined teachings of Liddelow I, Rubio, Williams, and Rivieccio render the invention unpatentable as claimed.
Claim 4 is rejected under 35 U.S.C. 103 as being unpatentable over Liddelow et al. (Nature, 2017, on IDS 01/15/2021, previously cited, hereafter “Liddelow I”) in view of Rubio et al. (Immunology, 2011), Williams et al. (PLoS One, 2009, on IDS 01/15/2021, previously cited) and Rivieccio et al. (The Journal of Immunology, 2005, previously cited), as evidenced by Foo et al. (Neuron, 2011, previously cited), as applied to claim 1 above, and further in view of Embad et al. (Stem cells and Development, 2012, previously cited).
In regards to claim 4, as discussed above, Liddelow I teaches that the astrocytes were obtained from forebrains, not pluripotent stem cells (PSCs).
However, a person of ordinary skill in the arts would have been motivated to derive astrocytes from PSCs because Embad teaches that embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), both types of PSCs, have the extraordinary ability to undergo both unlimited self-renewal and to differentiate into all cell types present in the adult organism, including specifically astrocytes, which are important as a source of therapeutic use for brain tumors and other neurological diseases (Abstract; Introduction, p404).
Furthermore, because Embad teaches methods for differentiating astrocytes from PSCs (Materials and Methods, p405; Figure 1, p406), it could have been done with predictable results and a reasonable expectation of success.
Therefore, the combined teachings of Liddelow 1, Rubio, Williams, Rivieccio, and Embad render the invention unpatentable as claimed.
Claim 5 is rejected under 35 U.S.C. 103 as being unpatentable over Liddelow et al. (Nature, 2017, on IDS 01/15/2021, previously cited, hereafter “Liddelow I”) in view of Rubio et al. (Immunology, 2011), Williams et al. (PLoS One, 2009, on IDS 01/15/2021, previously cited) and Rivieccio et al. (The Journal of Immunology, 2005, previously cited), as evidenced by Foo et al. (Neuron, 2011, previously cited), as applied to claim 1 above, and further in view of Liddelow et al. (Immunity Review, 2017, previously cited, hereafter “Liddelow II”).
In regards to claim 5, Liddelow I teaches that in previous experiments they found that neuroinflammation and ischemia induced two different types of reactive astrocytes that they termed A1 and A2 astrocytes (p481). In regards to the difference between these subsets, Liddelow I teaches that A1 astrocytes highly upregulate many classical complement cascade genes previously shown to be destructive to synapses, while A2 astrocytes upregulated many neurotrophic factors (p481). Therefore, Liddelow posits that A1 astrocytes are harmful, while A2 astrocytes are protective (p481).
This is confirmed by Liddelow II who teaches that injuries to the CNS elicit at least two types of “reactive” astrocytes, including A1 and A2 astrocytes (Abstract, p957; Transcriptome analysis of reactive astrocytes, p959). Liddelow II teaches the A1 and A2 states are a continuum of phenotype in response to inflammatory (A1 type) or ischemic (A2 type) states (Fig. 1, p959).
In comparison, Liddelow II teaches that physiologically, A1 astrocytes are involved in neuroinflammatory responses, and thus have “harmful” functions, while A2 astrocytes are induced by ischemia, promote survival and synapse repair, and thus have “helpful” functions (Transcriptome analysis of reactive astrocytes, p959).
In regards to the method of Liddelow I, in regards to the forebrain astrocytes as used by Liddelow I, as discussed above, Liddelow I teaches that the method for obtaining the astrocytes derives from Foo, and as evidenced by Foo, astrocytes obtained by this method strongly promote synapse formation (Summary, p799).
A person of ordinary skill in the art would have recognized that since A2 astrocytes promote synapse repair (formation), the astrocytes used by Liddelow I are A2 astrocytes specifically.
This is further suggested because as above, Liddelow II teaches that A2 and A1 astrocytes exist in a continuum, and develop an A1 phenotype in response to inflammation (Fig, 1, p959) including with inflammatory cytokine TNFα (p960-961).
However, even if the used cells as taught by Liddelow I were not A2 astrocytes specifically, a person of ordinary skill in the arts would have been motivated to use A2 astrocytes to test how the balance of A1 to A2 astrocytes affects human disease states, particularly neurodegenerative diseases such as Alzheimer’s disease, multiple sclerosis, ALS, Parkinson’s disease, and Huntington’s disease (p961-962; Fig, 1, p959).
Furthermore, because Liddelow II teaches that A1 astrocytes and A2 astrocytes are a continuum, and that A1 astrocytes can be activated by exposure to cytokines such as TNFα, as discussed above, it could have been done with predictable results and a reasonable expectation of success.
Therefore, the combined teachings of Liddelow 1, Rubio, Williams, Rivieccio, and Liddelow II renders the invention unpatentable as claimed.
Response to Arguments
Applicant argues that the claims require the specific concentration of TNFα of 0.5 ng/mL to 10 ng/mL, while Liddelow I teaches a concentration of 30 ng/mL (Remarks, p3).
Applicant’s arguments filed 06/10/2026 have been fully considered but are not found persuasive.
As discussed above, while a concentration of 30 ng/mL TNFα is greater than the claimed range of 0.5 to 10 ng/mL, a person of ordinary skill in the art could have arrived at a concentration of 0.5 to 10 ng/mL by routine optimization and the disclosure does not point to a criticality in this concentration range (see MPEP 2144.05(II)(A), Generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature 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) (Claimed process which was performed at a temperature between 40°C and 80°C and an acid concentration between 25% and 70% was held to be prima facie obvious over a reference process which differed from the claims only in that the reference process was performed at a temperature of 100°C and an acid concentration of 10%.); see also Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382 ("The normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine where in a disclosed set of percentage ranges is the optimum combination of percentages."); In re Hoeschele, 406 F.2d 1403, 160 USPQ 809 (CCPA 1969) (Claimed elastomeric polyurethanes which fell within the broad scope of the references were held to be unpatentable thereover because, among other reasons, there was no evidence of the criticality of the claimed ranges of molecular weight or molar proportions).
In the instant case, because Rubio teaches that astrocytes can be treated with 10 ng/mL TNFα in vitro for the production of pathological cytokines (Fig. 5, p366), a person of ordinary skill in the art could have arrived at the claimed concentration of 0.5 to 10 ng/mL by routine optimization with predicable results and a reasonable expectation of success.
Applicant argues that the Declaration filed under 37 CFR 1.132 provides evidence of patentability citing the figure (reproduced below) which shows neurite length when neurons are cultured on human astrocytes at varying concentrations (Remarks, p4; see also Declaration, p2). As argued by Applicant, “Shortened neurite length indicates that neurotoxicity was induced by A1 astrocytes” but that as shown in the figure “no neurotoxicity was induced by treatment with TNFα and IFNγ alone” (Remarks, p3). Continuing, Applicant argues that since TNFα and IFNγ alone showed no neurotoxicity, one of ordinary skill in the art could not conceive that TNFα and IFNγ could produce neurotoxic human A1 astrocytes (Remarks, p3).
Applicant’s arguments filed 06/10/2026 have been fully considered but are not found persuasive.
Neurotoxicty and shortening neurite length is an inherent property of A1 astrocytes themselves, not the specific result of contacting neurons with TNFα and IFNγ. As discussed above, Liddelow I explicitly teaches that A1 astrocytes are neurotoxic and kill neurons (and are thus cytotoxic) (p481) and that culturing A1 astrocytes with neurons results in shorter neurite outgrowth (Extended Data Figure 5k).
Thus, the A1 astrocytes of Liddelow I have the same properties as argued by Applicant as being the patentable feature.
Furthermore, Applicant’s arguments are not germane, because the claims are drawn to methods for “producing” A1 astrocytes not shortening neurite length or promoting cytotoxicity.
Thus, TNFα and IFNγ are not added because they cause have an effect on cytotoxicity or neurite length but because they have an effect on the production of A1 astrocytes.
As discussed above, Liddelow I explicitly teaches producing A1 astrocytes by culturing non-A1 astrocytes with TNFα.
While Liddelow I does not explicitly teach generating A1 astrocytes with TNFγ, as discussed above, a person of ordinary skill in the art would have been motivated to modify the method of Liddelow I and add IFNγ because Williams teaches that IFNγ and TNFα work together to promote CXCL10 expression in specifically a pro-inflammatory context (Abstract, p1; Figure 1, p3; p8), and as taught by Liddelow I, A1 astrocytes express CXCL10 (Figure 1, p482).
They would have been further motivated to add IFNγ because Williams also teaches that pro-inflammatory cytokines IFNγ and TNFα are elevated in the brains of patients with HIV-associated neurocognitive disorders (HAND,) such neuroinflammatory conditions such as HIV-encephalitis (HIVE) (Introduction, p1, p8), and a person of ordinary skill in the art would have been motivated to mimic the in vivo milieu of these patients in order to more accurately study human disease.
Moreover, they would have been motivated to add IFNγ because as taught by Rivieccio it promotes IL-1β-induced iNOS (thus, pro-inflammatory) expression (p3724).
Furthermore, because as above Liddelow I teaches that IFNγ promotes expression of A1 markers, because Williams demonstrates that the combination of IFNγ and TNFα promotes astrocyte CXCL10 expression in a pro-inflammatory context, and because Rivieccio teaches that astrocytes can be contacted with IFNγ in a pro-inflammatory context, it could have been done with predictable results and a reasonable expectation of success.
Applicant also argues that Test Example 1 demonstrates that adding TNFα and IFNγ in combination significantly increases C3 expression compared to C3 expression alone (citing Figs. 4 to 6), and that according to Test Example 3, the A1 astrocytes made by the instant method possess neurotoxicity (Remarks, p3).
Applicant argues that they do not need to specify a particular value for the expression level of C3 in the claims since the expression level of C3 demonstrates the effect of the claimed invention (Remarks, p5). Citing In re Merchant, 197 USPQ 785, 788 (CCPA 1978), Applicant argues that there is no law requiring that unexpected results relied upon for patentability be recited in the claims, and submits that the unexpected features related to the expression level of C3 need not be expressly recited in the claims since these noted advantages necessarily flow from the claimed invention as recited in the pending claims (Remarks, p5).
Applicant’s arguments filed 06/10/2026 have been fully considered but are not found persuasive.
In regards to allegations of unexpected results, Applicant is pointed to MPEP 716.02 which provides guidelines on assessing allegations of unexpected results.
Specifically, MPEP 716.02(d) states, “Whether the unexpected results are the result of unexpectedly improved results or a property not taught by the prior art, the ‘objective evidence of nonobviousness must be commensurate in scope with the claims which the evidence is offered to support.’ In other words, the showing of unexpected results must be reviewed to see if the results occur over the entire claimed range. In re Clemens, 622 F.2d 1029, 1036, 206 USPQ 289, 296 (CCPA 1980).”
In regards to In re Merchant, 197 USPQ 785, 788 (CCPA 1978), the MPEP discusses this reference not in the context of the analysis of whether the allegations are commensurate in scope with the claims as limited, but rather in regards to practices in regards to affidavits or declarations. Specifically, MPEP 716.02(e), states, “A comparison of the claimed invention with the disclosure of each cited reference to determine the number of claim limitations in common with each reference, bearing in mind the relative importance of particular limitations, will usually yield the closest single prior art reference." In re Merchant (emphasis in original).
Thus, Applicant’s arguments are not germane to the MPEP’s guidelines on assessing allegations of unexpected results.
Moreover, as admitted by Applicant “the unexpected features related to the expression level of C3 need not be expressly recited in the claims since these noted advantages necessarily flow from the claimed invention as recited in the pending claims” (see Remarks on 06/10/2026), and therefore, are an inherent property of A1 astrocytes as produced by the method steps.
Furthermore, Liddelow I teaches that C3 is one of the most characteristic and highly unregulated genes in A1 astrocytes (A1 astrocytes in human disease, p484), and as a result, high relative levels of C3 would have been the expected result.
Applicant argues that CXCL10 is not a marker specific to A1 astrocytes, but rather, is a pan-reactive marker (Remarks, p4). Applicant argues that the pan-reactivity of CXCL10 is supported by Wang et al. (Neuroscience, 2020) (Remarks, p4). Continuing, Applicant argue that even if Williams demonstrates that TNFα and IFNγ act synergistically to promote CXCL10 under pro-inflammatory conditions, Williams does not provide a proper reason, rationale, or motivation for using IFNγ to induce A1 astrocytes (Remarks, p4).
Applicant’s arguments filed 06/10/2026 have been fully considered but are not found persuasive.
In regards to Wang, if Applicant wishes Wang considered in regards to pan-reactivity then it needs to be submitted on an IDS.
However, in regards to a pan-specificity of CXCL10, CXCL10 is still an A1 astrocyte marker, and as acknowledged by Applicant, Williams teaches that IFNγ and TNFα work together to promote CXCL10 expression in a pro-inflammatory context (Abstract, p1; Figure 1, p3; p8) – which as taught by Liddelow I above is a specific A1 astrocyte context (and therefore, cellular milieu).
Therefore, while CXCL10 is expressed in both A1 and A2 subsets, because Williams teaches that IFNγ and TNFα work together to promote CXCL10 expression in a pro-inflammatory context (Abstract, p1; Figure 1, p3; p8) it is still a critical marker for A1 astrocytes specifically
Therefore, a person of ordinary skill in the art would have been motivated to modify the method of Liddelow I and add IFNγ because Williams teaches that IFNγ and TNFα work together to promote CXCL10 expression in specifically a pro-inflammatory context (Abstract, p1; Figure 1, p3; p8), and as taught by Liddelow I, A1 astrocytes express CXCL10 (Figure 1, p482).
They would have been further motivated to add IFNγ because Williams also teaches that pro-inflammatory cytokines IFNγ and TNFα are elevated in the brains of patients with HIV-associated neurocognitive disorders (HAND,) such neuroinflammatory conditions such as HIV-encephalitis (HIVE) (Introduction, p1, p8), and a person of ordinary skill in the art would have been motivated to mimic the in vivo milieu of these patients in order to more accurately study human disease.
Indeed, as further evidenced by Silva et al. (Journal of Inflammation, 2017), in other pro-inflammatory contexts, it was known in the art that IFNγ fuels astrocyte infection, promotes TNF, and results in a self-sustaining TNF-induced inflammatory milieu that perpetuates the parasite cycle and promotes cytokine-driven behavioral alterations (Title, Abstract, p1). Therefore, the pro-inflammatory cellular context of TNFα with IFNγ was well-established in the art.
Furthermore, because as above Liddelow I teaches that IFNγ promotes expression of A1 markers, because Williams demonstrates that the combination of IFNγ and TNFα promotes astrocyte CXCL10 expression in a pro-inflammatory context, it could have been done with predictable results and a reasonable expectation of success.
Applicant argues that “While A1 astrocytes can be produced when TNFα and IFNγ are used in combination, neurotoxic A1 astrocytes cannot be produced when TNFα or IFNγ are used alone (Remarks, p5). Applicant argues that although claim 1 is not a method for inducing neurotoxicity but rather relates to a method for producing A1 astrocytes that possess neurotoxicity, the content of the previously provided Declaration still relates to the claims and should be taken into consideration” (Remarks, p4).
Applicant’s arguments filed 06/10/2026 have been fully considered but are not found persuasive.
As discussed above, neurotoxicity (cytotoxicity) is known defining property of A1 astrocytes themselves. As discussed above, Liddelow I explicitly teaches that A1 astrocytes are neurotoxic and kill (p481).
Conclusion
No claims are allowed.
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/JOSEPH PAUL MIANO/Examiner, Art Unit 1631
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