Inflammasome antibody compositions and methods for treating neurological disorders

Abstract

Compositions and methods described herein include methods and agents for inhibiting inflammasome signaling in mammals, such as antibodies against inflammasome components, used alone or in combination with extracellular vesicle uptake inhibitors or other agents. Also described herein are compositions and methods of use for the detection and treatment of early Alzheimer's disease and other inflammatory neurological disorders.

Description

[Technical Field]

[0001] Description of electronically submitted text files The contents of the text file submitted electronically herewith are incorporated herein by reference in their entirety: Computer-readable format copy of the sequence listing (File name: IC100.xml, Recording date: September 4, 2023, File size: 54 kilobytes). [Background technology]

[0002] Background of the Invention The present invention relates generally to the fields of immunology and medicine. More specifically, the present invention relates to compositions and methods for modulating ASC (apoptosis-associated speck-like protein containing a caspase-activation recruitment domain (CARD)) activity and Absent in Melanoma 2 (AIM2) inflammasome activity in the mammalian central nervous system (CNS) and / or lung as a therapeutic approach to reduce inflammation in response to viral infection, injury, or conditions that cause inflammation in the CNS and / or lung. The present invention also relates to monoclonal antibodies or fragments thereof that specifically bind to ASC.

[0003] Alzheimer's disease (AD) is a progressive neurodegenerative disorder that disrupts memory and cognitive function. Inflammasome activation has been suggested to play a key role in the neuroinflammatory response during AD progression, but the cell-type expression of inflammasome proteins in the brain has not been fully characterized. Clinically, AD is characterized by functional and cognitive impairment, particularly memory impairment. Pathomechanisms contributing to AD include the accumulation of extracellular amyloid-β (Aβ) misfolded protein aggregates, intracellular hyperphosphorylated tau (pTau) neurofibrillary tangles, and chronic neuroinflammation. Anatomically, the hippocampus is affected early in the disease, followed by progression of neurodegeneration throughout the cerebral cortex.

[0004] Emerging evidence supports the idea that there is a link between misfolded protein aggregates and activation of the inflammasome of the innate immune system. Inflammasomes are composed of caspase-1, apoptosis-associated speck-like proteins containing a caspase recruitment domain (ASC), and sensor proteins such as NOD-like receptor 1 (NLRP1) and NLRP3. Sensor NLR proteins bind pathogen-associated molecular patterns (PAMPs) or danger-associated molecular patterns (DAMPs) to activate inflammasomes. Inflammasome activation involves sensor oligomerization, which then recruits monomeric ASC, which then oligomerizes with NOD-like receptors such as NLRP3 through a homotypic interaction between the PYRIN domain (PYD) of ASC and the PYD of NLRP3. Aggregation of ASC with NLRP3 recruits procaspase-1, which binds to ASC via a homotypic CARD-CARD interaction between the two proteins. Activated inflammasomes cleave caspase-1 to its active form, leading to the production of mature interleukin (IL)-1β and IL-18. These inflammatory cytokines are then secreted to propagate inflammatory signals. Active caspase-1 also cleaves gasdermin D (GSDM-D). Upon cleavage, the N-terminus of GSDM-D is inserted into the plasma membrane, forming a pore through which IL-1β and IL-18 are released, resulting in pyroptosis. Pyroptosis releases inflammasome proteins extracellularly, including the release of oligomerized ASC in the form of ASC specks that exhibit prion-like properties. Extracellular ASC specks persist for long periods while retaining their ability to cleave pro-IL-1β, thereby sustaining inflammation. Importantly, the propagation of prion-like inflammation by ASC specks contributes to a progressive inflammatory state that plays a central role in neurodegeneration.

[0005] Recent studies suggest that the NLRP3 inflammasome and ASC specks play important roles in the amplification of inflammatory responses and the aggregation of misfolded proteins characteristic of neurodegenerative diseases. For example, Aβ aggregates trigger NLRP-3 inflammasome activation in microglia. Importantly, extracellular ASC specks directly cross-seed Aβ aggregates in vitro and in vivo. Furthermore, therapeutics targeting the NLRP-3 inflammasome and ASC specks have shown promising results in animal models of AD.

[0006] Previous studies using anti-ASC antibodies have demonstrated that this treatment reduces pathology in several indications, including spinal cord injury, traumatic brain injury, acute lung injury, multiple sclerosis, and aging. We developed IC100, a humanized and deimmunized monoclonal antibody (mAb) (IgG4k) against ASC. In this study, we used IC100 and a commercially available anti-ASC antibody to determine the cell type distribution of ASC in postmortem brains from donors with AD. Furthermore, we used a panel of commercially available antibodies to identify Aβ, pTau, and the inflammasome proteins NLRP-1, NLRP-3, and caspase-1 in postmortem human brains with and without intermediate AD neuropathological changes. Severe acute respiratory syndrome coronavirus (SARS-CoV) is an enveloped virus with a single-stranded, positive-sense RNA genome (30 kb), which causes dangerous and potentially fatal conditions such as acute respiratory distress syndrome (ARDS) and acute lung injury (ALI). ARDS / ALI is a major public health problem, and despite significant advances in our understanding of the pathophysiology of this disease, mortality and morbidity rates remain high. According to the World Health Organization, the case fatality rate for SARS-CoV from November 2002 to July 2003 was 9.6%. While the case fatality rate for CoV-2 has yet to be precisely determined, coronaviruses are expected to recur cyclically, much like influenza. Therefore, there is a need to identify new treatments and vaccines to address current and future medical needs in this patient population. Summary of the Invention [Means for solving the problem]

[0007] Summary of the Invention In one aspect, provided herein is a monoclonal antibody or antibody fragment thereof that binds to an apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC), wherein the antibody or antibody fragment specifically binds to an epitope of ASC, wherein the epitope comprises or consists of the amino acid sequence of SEQ ID NO: 5, or 5-10, 10-15, or 15-20 amino acids of SEQ ID NO: 5.

[0008] In another aspect, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a heavy chain variable (VH) region and a light chain variable (VL) region, and the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3. Optionally, the VH region amino acid sequence comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22. Optionally, the ASC is a human ASC protein. Optionally, the antibody fragment is a Fab, F(ab')2, Fab', scFv, single domain antibody, diabody, or single chain camelid antibody or shark antibody. Optionally, the monoclonal antibody or antibody fragment thereof is human, humanized, or chimeric. Optionally, provided herein is an isolated nucleic acid molecule encoding the monoclonal antibody or antibody fragment thereof. Optionally, provided herein is an expression vector comprising the nucleic acid molecule. Optionally, provided herein is a nucleic acid molecule operably linked to a regulatory sequence suitable for expression of a nucleic acid segment in a host cell. Optionally, provided herein is a recombinant host cell comprising the expression vector. In another aspect, provided herein is a method for producing an antibody or antibody fragment that specifically binds to ASC, the method comprising culturing a recombinant host cell comprising the expression vector under conditions in which the nucleic acid molecule is expressed, thereby producing a monoclonal antibody or antibody fragment thereof that specifically binds to ASC. Optionally, provided herein is a pharmaceutical composition comprising the monoclonal antibody or antibody fragment thereof and a pharmaceutically acceptable carrier, diluent, or excipient. Optionally, provided herein is a method for treating inflammation in a subject, the method comprising administering a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof to the subject, thereby treating inflammation in the subject.In some embodiments, administration of the monoclonal antibody or antibody fragment thereof reduces at least the level of inflammatory cytokines. In some cases, the inflammation is inflammasome-associated inflammation. In some cases, the inflammasome-associated inflammation is associated with a viral infection, central nervous system (CNS) injury, an autoimmune disease, or a neurodegenerative disease. In some cases, the viral infection is caused by a coronavirus, such as SARS-CoV-2, MERS (Middle East Respiratory Virus) coronavirus, or an influenza virus, such as influenza A H5N1 (avian influenza) and influenza A H1N1 (swine influenza). In some cases, the CNS injury is selected from the group consisting of traumatic brain injury (TBI), stroke, and spinal cord injury (SCI). In some cases, the autoimmune disease or neurodegenerative disease is amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, muscular dystrophy (MD), or multiple sclerosis (MS).

[0009] In some cases, administration of the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject. In some cases, administration of the monoclonal antibody or antibody fragment thereof results in decreased ASC activity compared to a control. In some cases, the control is an untreated subject. In some cases, administration is intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation. In some cases, provided herein are methods for treating virus-associated pneumonia in a subject, including acute respiratory distress syndrome (ARDS), comprising administering a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof to the subject, thereby treating the pneumonia in the subject. In some cases, administration of the monoclonal antibody or antibody fragment thereof reduces the level of at least pro-inflammatory cytokines. In some cases, administration of the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject. In some cases, administration of the monoclonal antibody or antibody fragment thereof results in decreased ASC activity compared to a control. In some cases, the control is an untreated subject. Optionally, administration is intracerebroventricular, intraperitoneal, intravenous, intranasal, or by inhalation.

[0010] In yet another aspect, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, the antibody or antibody fragment comprising a light chain variable (VL) region and a heavy chain variable (VH) region, wherein the amino acid sequence of the VL region comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3. Optionally, the VL region amino acid sequence comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30, or 31. Optionally, the ASC is a human ASC protein. Optionally, the antibody fragment is a Fab, F(ab')2, Fab', scFv, single domain antibody, diabody, or single chain camelid antibody. Optionally, the monoclonal antibody or antibody fragment thereof is a human, humanized, or chimeric antibody or fragment. Optionally, provided herein is an isolated nucleic acid molecule encoding a monoclonal antibody or antibody fragment thereof. Optionally, provided herein is an expression vector comprising the nucleic acid molecule. Optionally, provided herein is a nucleic acid molecule operably linked to a regulatory sequence suitable for expression of a nucleic acid segment in a host cell. Optionally, provided herein is a recombinant host cell comprising the expression vector. In another aspect, provided herein is a method for producing an antibody or antibody fragment that specifically binds to ASC, the method comprising culturing a recombinant host cell comprising the expression vector under conditions in which the nucleic acid molecule is expressed, thereby producing a monoclonal antibody or antibody fragment thereof that specifically binds to ASC. Optionally, provided herein is a pharmaceutical composition comprising the monoclonal antibody or antibody fragment thereof and a pharmaceutically acceptable carrier, diluent, or excipient. Optionally, provided herein is a method for treating inflammation in a subject, the method comprising administering a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof to the subject, thereby treating inflammation in the subject.In some cases, administration of the monoclonal antibody or antibody fragment thereof reduces the level of at least inflammatory cytokines. In some cases, the inflammation is inflammasome-associated inflammation. In some cases, the inflammasome-associated inflammation is associated with central nervous system (CNS) injury, an autoimmune disease, or a neurodegenerative disease. In some cases, the CNS injury is selected from the group consisting of traumatic brain injury (TBI), stroke, and spinal cord injury (SCI). In some cases, the autoimmune disease or neurodegenerative disease is amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, muscular dystrophy (MD), or multiple sclerosis (MS). In some cases, administration of the monoclonal antibody or antibody fragment thereof inhibits inflammasome activation in the subject. In some cases, administration of the monoclonal antibody or antibody fragment thereof results in a decrease in ASC activity compared to a control. In some cases, the control is an untreated subject. In some cases, administration is intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation. Provided herein are methods for treating virus-associated pulmonary inflammation in a subject, including ARDS, comprising administering a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof to the subject, thereby treating MS in the subject. In some cases, administering the monoclonal antibody or antibody fragment thereof results in a reduction in the level of at least inflammatory cytokines. In some cases, administering the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject. In some cases, administering the monoclonal antibody or antibody fragment thereof results in a reduction in ASC activity compared to a control, where the control is an untreated subject. In some cases, administration is intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation.

[0011] In yet another aspect, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the amino acid sequence of the VH region comprises HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3; and the amino acid sequence of the VL region comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 or 22; and the VL region amino acid sequence comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 18, or an amino acid sequence at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 28, or an amino acid sequence at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 18, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 29, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 18, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 30, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30.In some cases, the VH region amino acid sequence comprises SEQ ID NO: 18, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 19, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 28, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 19, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 29, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 19, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 30, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 19, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31. In some cases, the VH region amino acid sequence comprises SEQ ID NO: 20, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 20; and the VL region amino acid sequence comprises SEQ ID NO: 28, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28.In some cases, the VH region amino acid sequence comprises SEQ ID NO:20, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:20; and the VL region amino acid sequence comprises SEQ ID NO:29, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:29. In some cases, the VH region amino acid sequence comprises SEQ ID NO:20, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:20; and the VL region amino acid sequence comprises SEQ ID NO:30, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:30. In some cases, the VH region amino acid sequence comprises SEQ ID NO:20, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:20; and the VL region amino acid sequence comprises SEQ ID NO:31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:31. In some cases, the VH region amino acid sequence comprises SEQ ID NO:21, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:21; and the VL region amino acid sequence comprises SEQ ID NO:28, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:28. In some cases, the VH region amino acid sequence comprises SEQ ID NO:21, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:21; and the VL region amino acid sequence comprises SEQ ID NO:29, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:29. In some cases, the VH region amino acid sequence comprises SEQ ID NO:21, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:21; and the VL region amino acid sequence comprises SEQ ID NO:30, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:30.In some cases, the VH region amino acid sequence comprises SEQ ID NO:21, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:21; and the VL region amino acid sequence comprises SEQ ID NO:31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:31. In some cases, the VH region amino acid sequence comprises SEQ ID NO:22, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:22; and the VL region amino acid sequence comprises SEQ ID NO:28, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:28. In some cases, the VH region amino acid sequence comprises SEQ ID NO:22, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:22; and the VL region amino acid sequence comprises SEQ ID NO:29, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:29. In some cases, the VH region amino acid sequence comprises SEQ ID NO:22, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:22; and the VL region amino acid sequence comprises SEQ ID NO:30, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:30. Optionally, the VH region amino acid sequence comprises SEQ ID NO:22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:22; and the VL region amino acid sequence comprises SEQ ID NO:31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:31. Optionally, the ASC is a human ASC protein. Optionally, the antibody fragment is a Fab, F(ab')2, Fab', scFv, single domain antibody, diabody, or single chain camelid antibody. Optionally, the monoclonal antibody or antibody fragment thereof is a human, humanized, or chimeric antibody or antibody fragment.Optionally, provided herein is an isolated nucleic acid molecule encoding a monoclonal antibody or antibody fragment thereof. In some aspects, provided herein is an expression vector comprising the nucleic acid molecule. Optionally, provided herein is a nucleic acid molecule operably linked to a regulatory sequence suitable for expression of a nucleic acid segment in a host cell. Optionally, provided herein is a recombinant host cell comprising the expression vector. In another aspect, provided herein is a method for producing an antibody or antibody fragment that specifically binds to ASC, the method comprising culturing a recombinant host cell comprising the expression vector under conditions in which the nucleic acid molecule is expressed, thereby producing a monoclonal antibody or antibody fragment thereof that specifically binds to ASC. Optionally, provided herein is a pharmaceutical composition comprising a monoclonal antibody or antibody fragment thereof and a pharmaceutically acceptable carrier, diluent, or excipient. Optionally, provided herein is a method for treating inflammation in a subject, the method comprising administering a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof to a subject, thereby treating inflammation in the subject. Optionally, administration of the monoclonal antibody or antibody fragment thereof reduces the level of at least a pro-inflammatory cytokine. Optionally, the inflammation is inflammasome-associated inflammation. In some cases, inflammasome-associated inflammation is associated with viral infection, central nervous system (CNS) injury, autoimmune disease, or neurodegenerative disease. In some cases, the viral infection is caused by a coronavirus such as SARS-CoV-2, or an influenza virus such as influenza A H5N1 (avian influenza) and influenza A H1N1 (swine influenza). In some cases, the CNS injury is selected from the group consisting of traumatic brain injury (TBI), stroke, and spinal cord injury (SCI). In some cases, the autoimmune disease or neurodegenerative disease is amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, muscular dystrophy (MD), or multiple sclerosis (MS). In some cases, administration of a monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in a subject.In some cases, administration of the monoclonal antibody or antibody fragment thereof results in a decrease in ASC activity compared to a control. In some cases, the control is an untreated subject. In some cases, administration is intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation. In some cases, a method of treating multiple sclerosis (MS) in a subject is provided, the method comprising administering a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof to the subject, thereby treating MS in the subject. ... , at least the level of inflammatory cytokines is reduced. In some cases, administration of the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject. In some cases, administration of the monoclonal antibody or antibody fragment thereof results in a decrease in ASC activity compared to a control. In some cases, the control is an untreated subject. In some cases, administration is intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation.

[0012] To demonstrate one aspect of the present invention, hippocampal formation, subiculum, and entorhinal cortex samples from the brains of 17 donors with low-level AD pathology and 17 donors with intermediate AD pathology were evaluated for inflammasome protein expression. Analysis of hippocampal thickness, β-amyloid plaques, and hyperphosphorylated tau was performed to identify cytopathological changes occurring between low and intermediate AD pathology. Next, changes in cells expressing the inflammasome sensor proteins NOD-like receptor protein (NLRP) 1 and 3, and caspase-1 were measured. Additionally, sections were stained with IC100, a humanized monoclonal antibody against the inflammasome adaptor protein apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and a commercially available anti-ASC antibody. The results showed no significant changes in hippocampal cortical thickness between low and intermediate AD pathology, but increased pTau and β-amyloid clusters were observed in intermediate AD cases. NLRP3 was primarily identified in microglial populations, whereas NLRP1 was found in neuronal plasmic regions. There was a significant increase in ASC in neurons labeled with IC100, whereas microglia in the hippocampus and subiculum were labeled with a commercially available anti-ASC antibody. Caspase-1 was present in the parenchyma of the CA region where amyloid and pTau were identified. Collectively, our results demonstrate that inflammasome protein expression increases in the early pathological stages of AD, that IC100 identifies early AD neurons, and that ASC expression correlates with Aβ and pTau in postmortem AD brains, suggesting that IC100 may be a promising therapeutic and diagnostic tool. [Brief explanation of the drawings]

[0013] [Figure 1A] Figure 1 shows inflammasome activation in C57 / BL6 mouse cortical and lung tissues after TBI. Figure 1A shows representative immunoblots of active caspase-1, ASC, IL-18, IL-β, HMGB1, and AIM2 after TBI. [Figure 1B] Active caspase-1 is significantly elevated in cortical tissue 4 and 24 hours after TBI. [Figure 1C] ASC is significantly elevated in cortical tissue 4 and 24 hours after TBI. [Figure 1D] IL-18 is significantly elevated in cortical tissue 4 and 24 hours after TBI. [Figure 1E] HMGB1 is significantly elevated in cortical tissue 4 and 24 hours after TBI. [Figure 1F] AIM2 is significantly elevated in cortical tissue 4 and 24 hours after TBI. [Figure 1G] IL-β was significantly elevated in cortical tissue at 4 and 24 hours after TBI. Data are shown as mean ± SEM. ***p<0.001, ***p<0.01, ***p<0.01, *p<0.05 compared to sham. N=4-5 / group. [Figure 1H] Figure 1H shows representative immunoblots for activated caspase-1, ASC, IL-18, IL-β, HMGB1, and AIM2 in lung tissue. [Figure 1I] Active caspase-1 is significantly elevated in lung tissue 4 and 24 hours after TBI. [Figure 1J] ASCs are significantly elevated in lung tissue 4 and 24 hours after TBI. [Figure 1K] IL-18 is significantly elevated in lung tissue 4 and 24 hours after TBI. [Figure 1L] HMGB1 is significantly elevated in lung tissue 4 and 24 hours after TBI. [Figure 1M] AIM2 is significantly elevated in lung tissue 4 and 24 hours after TBI. [Figure 1N] IL-β was significantly elevated in lung tissue 4 and 24 hours after TBI. Data are shown as mean ± SEM. N=4-5 / group. ***p<0.001, ***p<0.01, **p<0.01, *p<0.05 compared to sham. [Figure 2A]Figure 2 shows the expression of inflammasome proteins in type II alveolar epithelial cells. Figure 2A shows the increase of AIM2 in lung tissue after CCI (4 and 24 hours) compared to mice. [Figure 2B] Figure 2B shows an increase in active caspase-1 in lung tissue after CCI (4, 24 h) compared with mice. [Figure 2C] Figure 2C shows increased ASC immunoreactivity in lung tissue after CCI (4 and 24 hours) compared to mice. Confocal images of AIM2, caspase-1, and ASC (green) and type II epithelial cells (surfactant protein C, red). [Figure 3A] Figure 3 shows that TBI increases nuclear and cytoplasmic HMGB1 expression in mouse lungs. Figure 3A shows a representative immunoblot of nuclear HMGB1 after TBI. [Figure 3B] Figure 3B shows that nuclear HMGB1 is significantly elevated in 4-hour injured animals compared to sham. [Figure 3C] Figure 3C shows a representative immunoblot of cytoplasmic HMGB1 after TBI. [Figure 3D] Figure 3D shows that cytoplasmic HMGB1 is significantly elevated in 4-hour injured animals compared to sham. Data are shown as mean + / - SEM; *p<0.05 compared to sham. N=4-5 / group. [Figure 3E] Figure 3E shows increased HMGB1 immunoreactivity in lung tissue after CCI compared to sham mice. Confocal images of HMGB1 and type II epithelial cells (surfactant protein C, red). [Figure 4A] Figure 4 shows pyrosome formation in the lungs of mice 4 hours after TBI. Figure 4A demonstrates that TBI induces ASC laddering in lung tissue, indicating the formation of pyrosomes, i.e., oligomerization of ASC dimers that leads to caspase-1 activation and pyroptosis. [Figure 4B] Figure 4B shows a representative immunoblot. [Figure 4C]Figure 4C shows quantification of gasdermin D. Gasdermin D is significantly elevated in lung tissue after TBI. Data are shown as mean ± SEM. N = 4-5 per group. **p<0.01 compared to sham. [Figure 5A] Figure 5 shows that TBI induces alveolar morphological changes and acute lung injury in mice. Figure 5A shows H&E staining of lung sections from sham and injured animals at 4 and 24 hours. The sections show evidence of neutrophil infiltration (arrowheads), altered alveolar-capillary membrane morphology (asterisks, *), interstitial edema (short arrows), and thickening of the interstitium and alveolar septa (pounds, #). [Figure 5B] Figure 5B shows that acute lung injury scores were significantly increased in injured animals compared to sham at 4 and 24 hours. N = 4-5 / group, *p<0.05 / sham compared to sham. [Figure 6] Figure 6 shows the expression of CD81 in serum-derived EVs from control and TBI-injured mice. Representative immunoblots of CD81 in serum-derived EVs from sham control and TBI-injured mice. [Figure 7A] Figure 7 shows that adoptive transfer of EVs from TBI animals induces caspase-1 and ASC in the lungs of non-injured mice. Figure 7A shows representative immunoblots demonstrating that caspase-1 (Figure 7B), ASC (Figure 7C), IL-18 (Figure 7D), AIM2 (Figure 7E), and HMGB1 (Figure 7F) are elevated in the lungs of animals receiving EVs isolated from TBI mice compared to EVs from sham animals. Data are presented as mean ± SEM; *p<0.05 compared to sham. N=3 / group. [Figure 7B] We show that caspase-1 is elevated in the lungs of animals that received EVs isolated from TBI mice compared to EVs from sham animals. [Figure 7C] We show that ASCs are elevated in the lungs of animals that received EVs isolated from TBI mice compared with EVs from sham animals. [Figure 7D]We show that IL-18 is elevated in the lungs of animals that received EVs isolated from TBI mice compared to EVs from sham animals. [Figure 7E] We show that AIM2 is elevated in the lungs of animals that received EVs isolated from TBI mice compared to EVs from sham animals. [Figure 7F] We show that HMGB1 is elevated in the lungs of animals that received EVs isolated from TBI mice compared to EVs from sham animals. [Figure 7G] EVs from TBI mice induced morphological changes in alveoli (reduced alveolar size) and inflammatory cell infiltration, as determined by H&E staining. ALI scores were significantly increased in EVs delivered from injured mice compared to non-injured mice. Data are shown as mean ± SEM; **p<0.01, *p<0.05 compared to non-injured mice. [Figure 8A] Figure 8 shows that treatment with enoxaparin (3 mg / kg) and IC100 (5 mg / kg) reduces inflammasome expression in the lungs of animals receiving EVs from injured mice. Figure 8A shows representative immunoblots demonstrating that caspase-1 (Figure 8B), ASC (Figure 8C), IL-1β (Figure 8D), AIM2 (Figure 8E), and HMGB1 (Figure 8F) are reduced in the lungs of animals treated with enoxaparin and IC100 compared to untreated positive control animals. Data are presented as mean ± SEM; ***p<0.001, ***p<0.01, **p<0.01, *p<0.05 compared to sham. N=4 / group. [Figure 8B] Figure 1 shows that caspase-1 is reduced in the lungs of animals treated with enoxaparin and IC100 compared to untreated positive control animals. [Figure 8C] Figure 1 shows that ASC is reduced in the lungs of animals treated with enoxaparin and IC100 compared to untreated positive control animals. [Figure 8D] IL-1β is shown to be reduced in the lungs of animals treated with enoxaparin and IC 100 compared to untreated positive control animals. [Figure 8E] Figure 1 shows that AIM2 is decreased in the lungs of animals treated with enoxaparin and IC100 compared to untreated positive control animals. [Figure 8F] 1 shows that HMGB1 is decreased in the lungs of animals treated with enoxaparin and IC 100 compared to untreated positive control animals. [Figure 9A] Figure 9 shows that treatment with enoxaparin (3 mg / kg) and IC100 (5 mg / kg) reduces ALI scores in the lungs of animals receiving EVs from injured mice. Figure 9A shows H&E staining of a section of a saline-treated mouse lung. The section shows evidence of neutrophil infiltration, morphological changes in the alveolar-capillary membrane, interstitial edema, and thickening of the interstitium and alveolar septa. [Figure 9B] Figure 9B shows H&E staining of a section of an untreated mouse lung, showing evidence of neutrophil infiltration, morphological changes in the alveolar-capillary membrane, interstitial edema, and thickening of the interstitium and alveolar septa. [Figure 9C] Figure 9C shows H&E staining of a section of a mouse lung treated with enoxaparin (Figure 9C). The section shows evidence of neutrophil infiltration, morphological changes in the alveolar-capillary membrane, interstitial edema, and thickening of the interstitium and alveolar septa. [Figure 9D] Figure 9D shows H&E staining of lung sections from mice treated with IC 100 (anti-ASC). The sections show evidence of neutrophil infiltration, morphological changes in the alveolar-capillary membrane, interstitial edema, and thickening of the interstitium and alveolar septa. [Figure 9E] Figure 9E shows that acute lung injury scores were significantly reduced in animals treated with enoxaparin, IC100, compared to untreated animals. Data are presented as mean + / - SEM. N=4 / group, ****p<0.01, *p<0.05. [Figure 10A]Figure 10 shows that delivery of serum-derived EVs from TBI patients leads to increased inflammasome protein expression in lung endothelial cells. Figure 10A shows Western blot analysis of caspase-1, ASC, AIM2, and HMGB1 in PMVECs after 4 hours of incubation with TBI-EVs and control-EVs. [Figure 10B] Figure 10B shows quantification of Western blot, n=3 filters / group, n=6 patients, t-test, ****p<0.001, ***p<0.01, **p<0.01, *p<0.05. [Figure 10C] Figure 10C shows quantification of Western blot, n=3 filters / group, n=6 patients, t-test, ****p<0.001, ***p<0.01, **p<0.01, *p<0.05. [Figure 10D] Figure 10D shows quantification of Western blot, n=3 filters / group, n=6 patients, t-test, ****p<0.001, ***p<0.01, **p<0.01, *p<0.05. [Figure 10E] Figure 10E shows quantification of Western blot, n=3 filters / group, n=6 patients, t-test, ****p<0.001, ***p<0.01, **p<0.01, *p<0.05. [Figure 10F] Figure 10F shows immunoassay results of a significant increase in IL-1β expression using the Ella Simple Plex assay, n=3 filters / group, n=6 patients, t-test, ****p<0.001, ***p<0.01, **p<0.01, *p<0.05. [Figure 11A] Figure 11 shows that delivery of TBI-EVs to lung endothelial cells increases active caspase-1 immunoreactivity and cell death. Figure 11A shows caspase-1 FLICA colocalization and PI staining in PMVECs incubated with TBI-EVs for 4 hours. [Figure 11B] FIG. 11B shows caspase-1 FLICA and PI staining in PMVECs incubated with control-EVs for 4 hours. [Figure 11C]Figure 11C shows fluorescence plate reader analysis of PMVECs incubated with TBI and control-EVs for 4 hours, n=6, ***p<0.05. [Figure 12A] Figure 12 shows that treatment with a humanized anti-ASC monoclonal antibody (i.e., IC-100) improves functional outcomes in EAE. Figure 12A shows the clinical course of MOG35-55-induced EAE in C57BL / 6 mice treated with vehicle or increasing doses of IC-100. Administration of IC-100 (10, 30, and 45 mg / kg ip every 4 days) began on day 8, before the mice showed signs of paralysis. Results are expressed as the mean daily clinical score ± SEM for 9-10 mice per group. The 30 mg / kg and 45 mg / kg curves are significantly different from the vehicle curve; **p ≤ 0.001 Mann-Whitney test. [Figure 12B] FIG. 12B shows a comparison of peak clinical scores (highest disease score reached by a mouse) between groups; *p≦0.05, Student's t-test. [Figure 12C] Figure 12C shows a comparison of the cumulative disease index (CDI) between groups. CDI is equal to the sum of all scores from the day of onset for each animal and is a measure of EAE severity; *p<0.05, Student's t-test. [Figure 12D] Figure 12D shows a comparison of the onset date between groups, which is considered the day when mice first showed EAE symptoms. [Figure 12E] Figure 12E shows a comparison of peak disease days between groups. Peak disease day is the day when mice reach the highest disease score. Onset day is considered the day when mice show the first EAE symptoms. [Figure 13A] Figure 13 shows that IC-100 treatment reduces peripheral immune cell infiltration into the spinal cord after EAE. Figure 13A shows flow cytometry quantification of leukocyte populations infiltrating the spinal cord at 35 dpi after EAE. Results are expressed as mean ± SEM of 5 mice / group, *p<0.05, **p<0.001, Student's t-test. [Figure 13B]Figure 13B shows flow cytometry quantification of leukocyte populations present in the spleen at 35 dpi after EAE. Results are expressed as mean ± SEM of 5 mice / group, *p<0.05, **p<0.001, Student's t-test. [Figure 14] Figure 14 shows that IC-100 treatment reduces infiltration of peripheral immune cells into the spinal cord after EAE. Flow cytometry quantification of total microglia and MHCII-activated microglia in the spinal cord 35 dpi after EAE. Results are expressed as mean ± SEM of 5 mice / group, *p<0.05, Student's t-test. [Figure 15] FIG. 15 shows how IC-100 inhibits inflammasome formation and prevents the initiation of an immune response. [Figure 16] FIG. 16 shows how IC 100 inhibits ASC in ASC plaques and prevents inflammation from persisting. [Figure 17] Figure 17 shows an overview of the hippocampus and a schematic diagram of the strategy for quantification of Aβ, ptau, and inflammasome components. In A, the dotted lines delineate the boundaries of the hippocampal regions: dentate gyrus (DG); cornu ammonis 3 (CA3); cornu ammonis 2 (CA2); cornu ammonis 1 (CA1), and subiculum (subiculum). In B, the thickness of the hippocampal formation is compared between low and intermediate AD. Scale = 1 mm; ns = not significant; microns = μm. [Figure 18]Figure 18 shows AD neuropathological changes observed with Aβ and pTau. Aβ (A) and ptau (B) are shown in cases with Braak scores of 0-II and low AD neuropathological changes (low AD). AD neuropathological changes observed in intermediate AD cases with Braak scores of III-VI are shown (C: Aβ and D: pTau). E) Distribution of diffuse and neuritic Aβ plaques in intermediate AD cases compared to controls. F) Number of neurofibrillary tangles in the hippocampal regions of DG, CA1, CA2, CA3, and subiculum, and entorhinal cortex between intermediate AD cases compared to controls. Alzheimer's disease (AD), beta-amyloid (Aβ), hyperphosphorylated (ptau), dentate gyrus (DG); cornu ammonis 3 (CA3); cornu ammonis 2 (CA2); cornu ammonis 1 (CA1); subiculum (Sub); entorhinal cortex (EC); ns = not significant; * = p < 0.05; **** = p < 0.0001; Scale bar = 60 μm. [Figure 19] Figure 19 shows the localization of NLRP3 in microglia. NLRP3 immunoreactivity is shown in the CA1 (A and C) and CA2 (B and D) hippocampal regions in representative sections from a low-grade AD case (A and B) and an intermediate-grade AD case (C and D). NLRP3 expression is primarily observed in the processes of ramified microglia in the low-grade AD case (A, Ai and B, Bi), whereas in the intermediate AD case, expression is strongly stained in the processes of activated amoeboid microglia (C, Ci and D, Di). Alzheimer's disease (AD); NOD-like receptor protein (NLRP); ammonoid cortex 2 (CA2); ammonoid cortex 1 (CA1); Scale bar = 60 μm; inset scale bar = 30 μm. [Figure 20]Figure 20 shows NLRP1 expression in hippocampal neurons. NLRP1 immunoreactivity is shown in the CA1 (A and C) and CA2 (B and D) hippocampal regions of a case of low AD (A and B) and a case of intermediate AD (C and D). NLRP1 expression is more prevalent in neurons and apical dendrites in the CA1 region of a case of low AD (A) than in the CA2 (B) hippocampal region. In intermediate AD, NLRP1 immunoreactivity is observed in the form of clusters (blue arrows) in numerous neurons and parenchyma in CA1 (C) and CA2 (D). Alzheimer's disease (AD); NOD-like receptor protein (NLRP); cornu ammonis 2 (CA2); cornu ammonis 1 (CA1); Scale bar = 60 μm. [Figure 21] Figure 21 shows the difference in ASC expression between neurons and microglia. Mouse anti-ASC (A, C, and E) cell-type binding differs from that of IC100 in the hippocampal formation and entorhinal cortex (B, D, and F). Significant region-specific changes in cell counts between intermediate and low AD cases (E and F) are shown. Different morphologies of microglia are present at the CA1-CA2 border between low AD (A) and intermediate AD (C). Stereological analysis of cells stained with mouse anti-ASC was significantly higher in the DG, CA2, CA1, and Subregions in intermediate AD cases, whereas IC100 stained primarily neurons in the DG, CA3, CA2, and CA1 hippocampal regions in intermediate AD cases (D) compared with low AD (B). The caspase recruitment domain-containing adaptor protein apoptosis-associated speck-like protein (ASC); dentate gyrus (DG); cornu ammonis 3 (CA3); cornu ammonis 2 (CA2); cornu ammonis 1 (CA1); subiculum (Sub); and entorhinal cortex (EC). Scale bar = 60 μm; * = p < 0.05; ** = p < 0.01; *** = p < 0.001; ns = p > 0.05. [Figure 22]Figure 22 shows caspase-1 protein expression in the CA1 and CA2 hippocampal regions. The distribution of caspase-1 adjacent to neurons and in the tissue parenchyma is shown, resembling amyloid deposits. The CA1 (A) region of a low-AD case shows sparse clusters of caspase-1 near Aβ plaques (B), neurofibrillary tangles, and neurites (C). The CA2 region, a region less affected by neuropathological changes in low-AD, shows no clear caspase-1 immunoreactivity (D), no Aβ-positive plaques (E), but occasional neurofibrillary tangles (F). In an intermediate-AD case, both the CA1 (G) and CA2 (J) regions show dense clusters of caspase-1 expression, moderate density of Aβ plaques (H), and neurofibrillary tangles and neurites (J). β-amyloid (Aβ); hyperphosphorylated (ptau); cortical area 2 (CA2); cortical area 1 (CA1); Alzheimer's disease (AD); Scale bar = 30 μm. [Figure 23] Figure 23 provides supplemental data showing protein expression in tonsil and skin tissues. Protein expression of NLRP3, NLRP1, and caspase-1 in tonsils (A, C, and E) and skin (B, D, and F) is shown. Images A and B show NLRP3 expression in the germinal centers of tonsils (A) and epidermal cells of skin (B). NLRP1 expression is seen at low levels in the germinal centers of tonsils (C) and primarily in Langerhans cells of skin (D). Moderate expression of caspase-1 immunoreactivity was observed in tonsils (E) and skin (F). NOD-like receptor protein (NLRP). DETAILED DESCRIPTION OF THE INVENTION

[0014] Detailed Description Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0015] The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents cited herein, including, but not limited to, patents, patent applications, papers, books, and treatises, are expressly incorporated herein by reference in their entirety for any purpose. In the event that one or more of the incorporated documents or portions of documents defines a term that contradicts the definition of that term in this application, the definition set forth in this application shall control. However, reference to documents, papers, publications, patents, patent publications, and patent applications cited herein is not, and should not be construed as, an acknowledgment or any form of suggestion that they are valid prior art or form part of the general knowledge anywhere in the world.

[0016] The term "a" or "an" can refer to one or more of that entity, i.e., it can refer to a plural referent. Thus, the terms "a" or "an," "one or more," and "at least one" are used interchangeably herein. In addition, when referring to an "element" by the indefinite article "a" or "an," it does not exclude the possibility that there is more than one element, unless the context clearly requires that there is only one element.

[0017] Unless the context requires otherwise, throughout this specification and claims, "comprise" and variations thereof, such as "comprises" and "comprising," are to be interpreted in their open, inclusive sense, meaning "including, but not limited to." The use of alternatives (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the terms "about" and "consisting essentially of" mean + / - 20% of the stated range, value, or structure, unless otherwise indicated.

[0018] References throughout this specification to "one embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of the present disclosure. Thus, the appearances of the phrase "in one embodiment" or "in an embodiment" in various places throughout this specification do not necessarily all refer to the same embodiment. It will be understood that certain features of the present disclosure that are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the present disclosure that are described in the context of a single embodiment may also be provided separately or in any suitable subcombination.

[0019] Throughout this disclosure, various aspects of the methods and compositions provided herein may be presented in a range format. It should be understood that this description in range format is merely for convenience and brevity and should not be construed as an indefinite limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, and 3 to 6, as well as individual numerical values ​​within that range, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0020] As used herein, "protein" and "polypeptide" are used interchangeably to mean any peptide-linked chain of amino acids, regardless of length or post-translational modification, e.g., glycosylation or phosphorylation.

[0021] As used herein, the term "antibody" refers generally and broadly to immunoglobulin (Ig) molecules and immunologically active portions or fragments of immunoglobulin molecules, i.e., molecules that contain an antigen-binding site that specifically binds (immunoreacts with) an antigen (e.g., ASC, NLRP1, AIM2, etc.). The antibodies provided herein can be polyclonal antibodies, monoclonal antibodies (mAbs), chimeric antibodies, humanized antibodies, anti-idiotypic (anti-Id) antibodies, antibodies that can be labeled in soluble or conjugated form, as well as active fragments, regions, or derivatives thereof. As used herein, antibodies can be chimeric, humanized, or human antibodies.

[0022] "Specifically binds" or "immunoreacts" means that an antibody reacts with one or more antigenic determinants of a desired antigen and does not react with other polypeptides. In certain embodiments, an antibody is said to specifically bind to an antigen if it preferentially recognizes its target antigen in a complex mixture of proteins and / or macromolecules. The term "antibody" generally refers broadly to an immunoglobulin (Ig) molecule, which generally comprises four polypeptide chains: two heavy (H) chains and two light (L) chains, or a functional fragment, mutant, variant, or derivative thereof that retains the essential target-binding characteristics of an Ig molecule. Such mutant, variant, or derivative antibody formats are known in the art. Such anti-ASC and anti-NLRP1 antibodies of the present invention can bind to portions of ASC and NLRP1, respectively, that inhibit caspase-1 activation.

[0023] As used herein, the term "humanized antibody" refers to an antibody in which minimal portions of an otherwise non-human antibody have been introduced into the human antibody.

[0024] As used herein, the term "human antibody" refers to an antibody in which substantially all portions of the protein have only minor sequence changes or mutations, making them substantially non-immunogenic in humans.

[0025] In a full-length antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains: CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain, CL. The VH and VL regions are further subdivided into regions of hypervariability called complementarity-determining regions (CDRs), which are separated by highly conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) and class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) or subclass. IgG, IgD, and IgE antibodies generally comprise two identical heavy chains and two identical light chains, and two antigen-binding domains, each consisting of a heavy-chain variable region (VH) and a light-chain variable region (VL). IgA antibodies generally comprise two monomers, each consisting of two heavy chains and two light chains (as with IgG, IgD, and IgE antibodies); thus, an IgA molecule has four antigen-binding domains, each also consisting of a VH and a VL. Certain IgA antibodies are monomers consisting of two heavy chains and two light chains. Secreted IgM antibodies are generally composed of five monomers, each consisting of two heavy chains and two light chains (similar to IgG and IgE antibodies); thus, an IgM molecule has 10 antigen-binding domains, each also consisting of a VH and VL. Cell surface forms of IgM also exist, which have a two-heavy / two-light chain structure similar to IgG, IgD, and IgE antibodies.

[0026] As used herein, the terms "antigen-binding fragment" or "antigen-binding portion" or "antigen-binding site" or "binding domain" or "binding region" can refer to a domain, region, portion, or portion of a protein, polypeptide, oligopeptide, or peptide, or an antibody or antibody-derived binding domain, that retains the ability to specifically bind to an antigen (e.g., an ASC protein). Exemplary binding domains include single-chain antibody variable regions (e.g., domain antibodies, sFv, scFv, scFab), fusion proteins comprising antibody portions (e.g., domain antibodies), receptor ectodomains, and ligands (e.g., cytokines, chemokines). In one embodiment, the fusion protein comprises one or more CDRs. In another embodiment, the fusion protein comprises CDR H3 (VH CDR3) and / or CDR L3 (VL CDR3). For purposes of this invention, a fusion protein comprises one or more antibodies and additional amino acid sequences, such as heterologous sequences or homologous sequences from other regions, attached to the N- or C-terminus of the antibody or antibody fragment thereof. Exemplary heterologous sequences include, but are not limited to, "tags" such as a FLAG tag or a 6His tag, or enzymes or polypeptides that increase the half-life of an antibody in the blood. Tags are known in the art. The additional amino acid sequences that may comprise amino- and / or carboxyl-terminal fusions can vary in length from one residue to over 100 residues, and even polypeptides containing intrasequence insertions of single or multiple amino acid residues.

[0027] An antigen-binding site is generally formed by heavy chain variable region (VH) and light chain variable region (VL) immunoglobulin domains, and the antigen-binding interface is formed by six surface polypeptide loops called complementarity-determining regions (CDRs). Three CDRs are present in each of the VH (HCDR1, HCDR2, HCDR3) and VL (LCDR1, LCDR2, LCDR3), along with framework regions (FR). In certain embodiments, the binding domain comprises or consists of an antigen-binding site (e.g., comprising variable heavy and variable light chain sequences or three light chain complementarity-determining regions (CDRs) and three heavy chain CDRs from an antibody arranged in alternative framework regions (FRs) (e.g., human FRs, optionally containing one or more amino acid substitutions)).

[0028] The term "CDR region" or "CDR" can refer to the hypervariable region of an immunoglobulin heavy or light chain as defined by Kabat et al., 1991 (Kabat, EA et al., (1991) Sequences of Proteins of Immunological Interest, 5th Edition. U.S. Department of Health and Human Services, Public Service, NIH, Washington) and subsequent editions. Antibodies typically contain three heavy chain CDRs and three light chain CDRs.

[0029] It has been demonstrated that the antigen-binding function of an antibody can also be performed by fragments of a full-length antibody. Antibody and antibody fragment embodiments may also be in a bispecific, trispecific, dual-specific, or multispecific format; specifically binding to two or more different antigens. Examples of binding fragments encompassed by the term "antigen-binding fragment" of an antibody include: (i) a Fab fragment consisting of the VL, VH, CL, and CH1 domains (Ward, ES et al., (1989) Nature 341, 544-546); (ii) a Fd fragment consisting of the VH and CH1 domains (McCafferty et al., (1990) Nature, 348, 552-554); (iii) a Fv fragment consisting of the VL and VH domains of a single antibody (Holt et al., (2003) Trends in Biotechnology 21, 484-490); (iv) a dAb fragment consisting of the VH or VL domain (Ward, ES et al., Nature 341, 544-546 (1989), McCafferty et al., (1990) Nature, 348, 552-554, Holt et al., (2003) Trends in Biotechnology 21, 484-490]; (v) isolated CDR regions; (vi) F(ab')2 fragments, bivalent fragments comprising two linked Fab fragments; and (vii) single-chain Fv molecules (scFv), in which the VH and VL domains are linked by a peptide linker that allows the two domains to associate and form an antigen-binding site (Bird et al., (1988) Science, 242, 423-426; Huston et al., (1988) PNAS USA, 85, 5879-5883). The present invention also encompasses Fab' fragments.Furthermore, the two domains of an Fv fragment, VL and VH, are encoded by separate genes but can be recombinantly linked by a synthetic linker that allows the VL and VH regions to pair and form a single protein chain (known as a single-chain Fv (scFv)) to form a monovalent molecule. Such single-chain antibodies are also intended to be encompassed by the term "antigen-binding fragment" of an antibody. In certain embodiments of the invention, scFv molecules can be incorporated into fusion proteins. In some embodiments, the invention includes single-chain camelid antibodies; (viii) bispecific single-chain Fv dimers (PCT / U.S. Pat. No. 92,109,965); and (ix) "diabodies," multivalent or multispecific fragments constructed by gene fusion (WO 94 / 13804; Holliger, P. (1993) et al., Proc. Natl. Acad. Sci. USA 90 6444-6448). Diabodies are bivalent, bispecific antibodies in which a VH domain and a VL domain are expressed on a single polypeptide chain, but these domains are forced to pair with complementary domains on another chain by using a linker that is too short to allow pairing between the two domains on the same chain, thereby creating two antigen-binding sites (see, e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1121-1123). Such antibody-binding fragments are known in the art (Kontermann and Dubel eds., Antibody Engineering (2001) Springer-Verlag. New York. 790 pp.). In some embodiments, the present invention includes single-domain antibodies. In general, as used herein, the term "antibody" encompasses "antibody fragments." Antibody fragments generally retain antigen-binding properties of the full-length antibody.

[0030] Fv, scFv, or diabody molecules can be stabilized by incorporating disulfide bridges linking the VH and VL domains (Reiter, Y. et al., Nature Biotech, 14, 1239-1245, 1996). Minibodies containing scFvs linked to CH3 domains can also be produced (Hu, S. et al., (1996) Cancer Res., 56, 3055-3061). Other examples of binding fragments are Fab', which differs from Fab fragments by the addition of a few residues to the carboxyl terminus of the heavy chain CH1 domain, such as one or more cysteines from the antibody hinge region, and Fab'-SH, a Fab' fragment in which the cysteine ​​residues of the constant domains bear free thiol groups.

[0031] As used herein, "Fv" can refer to the minimum antibody fragment that retains both the antigen recognition and binding sites. As used herein, "Fab" can refer to an antibody fragment that contains the constant domain of the light chain and the CH1 domain of the heavy chain. The term "mAb" refers to a monoclonal antibody.

[0032] "Fc region" or "Fc domain" refers to a polypeptide sequence corresponding to or derived from the portion of a source antibody responsible for binding to antibody receptors on cells and the C1q component of complement. Fc refers to "fragment crystalline," i.e., a fragment of an antibody that readily forms protein crystals. Individual protein fragments were initially described by proteolytic digestion, but the overall general structure of immunoglobulin proteins can be defined. As originally defined in the literature, an Fc fragment consists of the disulfide-linked heavy chain hinge region, CH2, and CH3 domains. However, more recently, the term has been used to refer to a single chain consisting of the CH3, CH2, and at least a portion of the hinge sufficient to form a disulfide-linked dimer with a second such chain. For a general overview of immunoglobulin structure and function, see Putnam, The Plasma Proteins, Vol. V (Academic Press, Inc., 1987), pp. 49-140; and Padlan, Mol. Immunol. 31:169-217, 1994. As used herein, the term Fc includes naturally occurring sequence variants. In one embodiment, the antibodies or antibody fragments derived therefrom (e.g., anti-ASC monoclonal antibodies or antibody fragments thereof) provided herein have a modified Fc region or domain. In some cases, the modified Fc region or domain can confer improved thermal stability to the resulting antibody or antibody fragment derived therefrom. Improved thermal stability can result in increased serum half-life. The Fc region or domain can be modified as described in U.S. Patent Application Publication No. 20160193295, the contents of which are incorporated herein by reference. As described in U.S. Patent Application Publication No. 20160193295, the Fc region or domain can be modified to have one or more cysteine ​​residues in the hinge region deleted and one or more CH3-interface amino acids substituted with sulfhydryl-containing residues.In another embodiment, the Fc region or domain of an antibody provided herein or antibody fragment thereof (e.g., an anti-ASC monoclonal antibody or antibody fragment thereof) can be stabilized by engineering the Fc region to have intradomain disulfide bonds, as described in Wozniak-Knopp G, Stadlmann J, Rueker F (2012) Stabilization of the Fc Fragment of Human IgG1 by Engineered Intradomain Disulfide Bonds. PLoS ONE 7(1): e30083, the contents of which are incorporated herein by reference. In yet another embodiment, the antibody has an Fc region modified as described in WO 99 / 58572, the contents of which are incorporated herein by reference. In yet another embodiment, the Fc region or domain can be modified as described in U.S. Pat. No. 9,574,010, the contents of which are incorporated herein by reference.

[0033] The terms "apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (CARD)" and "ASC" refer to the expression product of the ASC gene or an isoform thereof, or a protein that shares at least 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with ASC (e.g., human NP_037390 (Q9ULZ3-1), NP_660183 (Q9ULZ3-2), or Q9ULZ3-3, mouse NP_075747, or rat NP_758825 (BAC43754)), and exhibits the functional activity of ASC. A "functional activity" of a protein is any activity related to the physiological function of the protein. Functional activity of ASC includes, for example, activation of caspase-1 and recruitment of proteins for the initiation of cell death.

[0034] The term "ASC gene" or "ASC nucleic acid" refers to a native ASC-encoding nucleic acid sequence, a genomic sequence from which an ASC cDNA can be transcribed, and / or allelic variants and homologs thereof. The term encompasses double-stranded DNA, single-stranded DNA, and RNA.

[0035] As used herein, the term "inflammasome" refers to a multiprotein (e.g., at least two protein) complex that activates caspase-1. Furthermore, the term "inflammasome" can refer to a multiprotein complex that activates caspase-1 activity, which in turn regulates the processing and activation of IL-1β, IL-18, and IL-33. See Arend et al. 2008; Li et al. 2008; and Martinon et al. 2002 (each incorporated herein by reference). The term "NLRP1 inflammasome," "NALP1 inflammasome," "NLRP2 inflammasome," "NALP2 inflammasome," "NLRP3 inflammasome," "NALP3 inflammasome," "NLRC4 inflammasome," "IPAF inflammasome," or "AIM2 inflammasome" refers to a protein complex of at least caspase-1 and one adaptor protein, such as ASC. For example, the terms "NLRP1 inflammasome" and "NALP1 inflammasome" can refer to a multiprotein complex containing NLRP1, ASC, caspase-1, caspase-11, XIAP, and pannexin-1 for activating caspase-1 and processing interleukin-1β, interleukin-18, and interleukin-33. The terms "NLRP2 inflammasome" and "NALP2 inflammasome" can refer to a multiprotein complex containing NLRP2 (also known as NALP2), ASC, and caspase-1. The terms "NLRP3 inflammasome" and "NALP3 inflammasome" can refer to a multiprotein complex containing NLRP3 (also known as NALP3), ASC, and the terms "NLRC4 inflammasome" and "IPAF inflammasome" can refer to a multiprotein complex containing NLRC4 (also known as IPAF), ASC, and caspase-1. Furthermore, the term "AIM2 inflammasome" can refer to a multiprotein complex containing AIM2, ASC, and caspase-1.

[0036] As used herein, the phrase "sequence identity" refers to the percentage of identical subunits at corresponding positions in two sequences (e.g., nucleic acid sequences, amino acid sequences) when the two sequences (e.g., nucleic acid sequences, amino acid sequences) are aligned to maximize subunit correspondence, i.e., taking into account gaps and insertions. Sequence identity can be measured using sequence analysis software (e.g., Sequence Analysis Software Package from Accelrys CGC, San Diego, Calif.).

[0037] The phrases "therapeutically effective amount" and "effective dosage" refer to an amount sufficient to produce a therapeutically (e.g., clinically) desired result; the specific result may vary depending on the nature (type) of the disorder being treated. For example, when the disorder being treated is SCI, the result may be improved motor skills and function, reduced spinal cord pathology, etc. The compositions described herein may be administered one or more times daily to one or more times weekly. One of skill in the art will appreciate that certain factors, including but not limited to, the severity of the disease or disorder, previous treatments, the subject's general health and / or age, and other diseases present, may influence the dosage and timing required to effectively treat a subject. Furthermore, treatment of a subject with a therapeutically effective amount of a composition of the invention may include a single treatment or a series of treatments.

[0038] As used herein, the term "treatment" is defined as the application or administration of a therapeutic agent described herein, or identified by the methods described herein, to a patient having a disease, a symptom of a disease, or a predisposition to a disease, or to a tissue or cell line isolated from a patient, for the purpose of curing, ameliorating, alleviating, mitigating, altering, eliminating, ameliorating, or affecting the disease, symptom of a disease, or predisposition to a disease.

[0039] As used herein, the terms "patient," "subject," and "individual" are used interchangeably and refer to a mammalian subject to be treated. In one embodiment, the mammalian patient is a human. In some cases, the methods of the present invention are useful in experimental animals, veterinary applications, and the development of disease model animals, including, but not limited to, rodents such as mice, rats, and hamsters, and primates.

[0040] As used interchangeably herein, "Absent in Melanoma 2" and "AIM2" can refer to the expression product of the AIM2 gene or isoform; or a protein that shares at least 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with AIM2 and exhibits the functional activity of AIM2 (e.g., accession numbers NX_014862, NP004824, XP016858337, XP005245673, AAB81613, BAF84731, AAH10940).

[0041] As used interchangeably herein, "NACHT, LRR and PYD domain-containing protein 1," "NALP1," and "NLRP1" refer to the expression product of the NALP1 or NLRP1 gene or isoform; or a protein that shares at least 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with NALP1 and exhibits the functional activity of NALP1 (e.g., Accession Nos. AAH51787, NP_001028225, NP_127500, NP_127499, NP_127497, NP055737).

[0042] As used interchangeably herein, "NALP2" and "NLRP2" refer to the expression product of the NALP2 or NLRP2 gene or isoform; or a protein that shares at least 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with NALP2 and exhibits the functional activity of NALP2 (e.g., accession numbers NP_001167552, NP_001167553, NP_001167554, or NP_060322).

[0043] As used interchangeably herein, "NALP3" and "NLRP3" refer to the expression product of the NALP3 or NLRP3 gene or isoform; or a protein that shares at least 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with NALP3 and exhibits the functional activity of NALP3 (e.g., accession numbers NP_001073289, NP_001120933, NP_001120934, NP_001230062, NP_004886, NP_899632, XP_011542350, XP_016855670, XP_016855671, XP_016855672, or XP_016855673).

[0044] As used interchangeably herein, "NLRC4" and "IPAF" refer to the expression product of the NLRC4 or IPAF gene or isoform; or a protein that shares at least 65%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with NLRC4 and exhibits the functional activity of NLRC4 (e.g., accession numbers NP_001186067, NP_001186068, NP_001289433, or NP_067032).

[0045] The terms "stroke" and "ischemic stroke" refer to when blood flow to part of the brain or spinal cord is interrupted.

[0046] "Traumatic injury to the CNS" means any injury to the CNS due to an external mechanical force that can result in permanent or temporary impairment of CNS function.

[0047] The methods described herein include conventional molecular biology techniques.Such techniques are generally known in the art and are described in detail in methodological textbooks such as: Molecular Cloning: A Laboratory Manual, 3rd ed., vol. 1-3, ed. Sambrook et al., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2001; and Current Protocols in Molecular Biology, ed. Ausubel et al., Greene Publishing and Wiley-Interscience, New York, 1992 (regularly updated). Immunological techniques are generally known in the art and are described in detail in methodological treatises such as, for example, Advances in Immunology, volume 93, ed. Frederick W. Alt, Academic Press, Burlington, Mass., 2007; Making and Using Antibodies: A Practical Handbook, eds. Gary C. Howard and Matthew R. Kaser, CRC Press, Boca Raton, Fla., 2006; Medical Immunology, 6th ed., edited by Gabriel Virella, Informa Healthcare Press, London, England, 2007; and Harlow and Lane ANTIBODIES: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988.

[0048] Although compositions and methods similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable compositions and methods are described below. All publications, patent applications, and patents mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. The specific embodiments described below are illustrative only and are not intended to be limiting.

[0049] Provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to an apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC). The monoclonal antibody or fragment thereof is capable of specifically binding to an antigenic fragment of ASC comprising, consisting of, or consisting essentially of the amino acid sequence of SEQ ID NO:5. In addition to this embodiment, the present invention also contemplates the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. In one embodiment, the monoclonal antibody or antibody fragment thereof provided herein can be used in a method for reducing inflammation in a mammal, as described in U.S. Pat. No. 8,685,400, the contents of which are incorporated herein by reference in their entirety. The inflammation may be in the lungs and / or central nervous system (CNS). Inflammation of the lungs and / or CNS can be the result of infection (viral or bacterial), injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or affliction of or affecting the CNS. Viral infection can be caused by a coronavirus, such as SARS-CoV-2, or an influenza virus, such as influenza A H5N1 (avian influenza) and influenza A H1N1 (swine influenza). As provided herein, the disease, condition, or affliction of the CNS or a disease, condition, or affliction affecting the CNS can be stroke, and autoimmune and / or CNS diseases, including amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The monoclonal antibodies and fragments thereof described herein can also be used prophylactically in methods for preventing inflammation associated with any of the disorders disclosed herein.

[0050] Patients treated with the methods of the invention may be experiencing a hyperinflammatory response, also known as cytokine storm syndrome, a secondary hemophagocytic lymphohistiocytosis (sHLH) characterized by fulminant hypercytokinemia leading to multiple organ failure (Mehta, McAuley et al. 2020). The antibodies, antibody fragments, and methods of the invention can be used to treat or prevent hyperinflammation or cytokine storm syndrome associated with viral infections, including hyperinflammation and cytokine storm associated with SARS-CoV-2 infection.

[0051] The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. This reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In some embodiments, the antibodies and fragments thereof used in the methods described herein bind to and inhibit the ASC portion of the inflammasome, thereby preventing assembly of the multiprotein inflammasome, which is necessary for the initiation of an inflammatory response (Figure 15). The antibodies and fragments thereof can also bind to and inhibit ASC specks or ASC monomers, both intracellularly and extracellularly. This inhibits propagation of the large filamentous signaling platform and prevents the perpetuation of inflammation associated with chronic inflammatory diseases mediated by the ASC-dependent inflammasome (Figure 16).

[0052] In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat pneumonia associated with a viral infection by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from viral-associated pneumonia, including ARDS and / or ALI. The monoclonal antibody or antibody fragment thereof of this embodiment can be present in a composition, such as a pharmaceutical composition as provided herein. In some cases, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agents may be any of the agents provided herein (e.g., EV uptake inhibitors) and / or antibodies or antibody fragments against other inflammasome components (e.g., IL-18, caspase-1, NLRP1, NLRP3, AIM2, etc.). In some cases, the other agents are hydroxychloroquine, chloroquine, and immunosuppressants, including, but not limited to, steroids, selective cytokine blockers (e.g., anakinra or tocilizumab), JAK inhibitors, interleukin inhibitors including IL-1β and IL-6 inhibitors, TNF inhibitors, and CSF inhibitors, and anticoagulants such as low molecular weight heparin, tissue plasminogen activator, or enoxaparin.

[0053] In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. The monoclonal antibody or antibody fragment thereof of this embodiment may be present in a composition, such as a pharmaceutical composition as provided herein. In some cases, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agents may be any of the agents provided herein (e.g., EV uptake inhibitors) and / or antibodies or antibody fragments against other inflammasome components (e.g., IL-18, caspase-1, NALP1, AIM2, etc.).

[0054] The present invention also encompasses a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a heavy chain variable (VH) region and a light chain variable (VL) region, and the amino acid sequence of the VH region comprises HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3. Furthermore, in this embodiment, the present invention contemplates the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. In one embodiment, the monoclonal antibody or antibody fragment thereof provided herein can be used in a method for reducing inflammation in a mammal, as described in U.S. Patent No. 8,685,400, the contents of which are incorporated herein by reference in their entirety. The inflammation may be in the lungs and / or CNS. Inflammation in the lungs and / or CNS can be the result of viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or affliction of the CNS or a disease, condition, or affliction affecting the CNS. As provided herein, the disease, condition, or affliction of the CNS or a disease, condition, or affliction affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce innate immune or inflammasome-associated inflammation in a patient. The reduction may be relative to a control (eg, an untreated patient and / or a patient prior to treatment).In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat pneumonia associated with a viral infection by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from viral-associated pneumonia, including ARDS and / or ALI. The monoclonal antibody or antibody fragment thereof of this embodiment may be present in a composition, such as a pharmaceutical composition as provided herein. In some cases, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agents may be any of the agents provided herein (e.g., EV uptake inhibitors) and / or antibodies or antibody fragments against other inflammasome components (e.g., IL-18, caspase-1, NLRP1, NLRP3, AIM2, etc.). Optionally, the other agents include hydroxychloroquine, chloroquine, and immunosuppressants, including, but not limited to, steroids, selective cytokine blockers (e.g., anakinra or tocilizumab), JAK inhibitors, interleukin inhibitors including IL-1β and IL-6 inhibitors, TNF inhibitors, and CSF inhibitors, as well as anticoagulants such as low molecular weight heparin, tissue plasminogen activator, or enoxaparin. In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. The monoclonal antibody or antibody fragment thereof of this embodiment can be present in a composition, such as, for example, a pharmaceutical composition as provided herein. Optionally, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agent may be any agent provided herein (e.g., an EV uptake inhibitor) and / or an antibody or antibody fragment against other inflammasome components (e.g., IL-18, caspase-1, NALP1, AIM2, etc.).

[0055] In some embodiments, the present invention provides a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a light chain variable (VL) region and a heavy chain variable (VH) region, and the amino acid sequence of the VL region comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3. Further to this embodiment, the present invention contemplates the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. In one embodiment, the monoclonal antibody or antibody fragment thereof provided herein can be used in a method for reducing inflammation in a mammal, as described in U.S. Patent No. 8,685,400, the contents of which are incorporated herein by reference in their entirety. The inflammation may be in the lungs and / or CNS. Inflammation in the lungs and / or CNS can be the result of viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or affliction of the CNS or a disease, condition, or affliction affecting the CNS. As provided herein, the disease, condition, or affliction of the CNS or a disease, condition, or affliction affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce innate immune or inflammasome-associated inflammation in a patient. The reduction may be relative to a control (eg, an untreated patient and / or a patient prior to treatment).In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat pneumonia associated with a viral infection by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from viral-associated pneumonia, including ARDS and / or ALI. The monoclonal antibody or antibody fragment thereof of this embodiment may be present in a composition, such as a pharmaceutical composition as provided herein. In some cases, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agents may be any of the agents provided herein (e.g., EV uptake inhibitors) and / or antibodies or antibody fragments against other inflammasome components (e.g., IL-18, caspase-1, NLRP1, AIM2, etc.). Optionally, the other agents are hydroxychloroquine, chloroquine, and immunosuppressants, including, but not limited to, steroids, selective cytokine blockers (e.g., anakinra or tocilizumab), JAK inhibitors, interleukin inhibitors such as IL-1β and IL-6 inhibitors, TNF inhibitors, and CSF inhibitors, as well as anticoagulants such as low molecular weight heparin, tissue plasminogen activator, or enoxaparin. In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. The monoclonal antibody or antibody fragment thereof of this embodiment can be present in a composition, such as, for example, a pharmaceutical composition as provided herein. Optionally, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agent may be any agent provided herein (e.g., an EV uptake inhibitor) and / or an antibody or antibody fragment against other inflammasome components (e.g., IL-18, caspase-1, NALP1, AIM2, etc.).

[0056] In another embodiment, the present invention also provides a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region amino acid sequence comprises an HCDR1 of SEQ ID NO: 6, an HCDR2 of SEQ ID NO: 7, and an HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3; and the VL region amino acid sequence comprises an LCDR1 of SEQ ID NO: 12, an LCDR2 of SEQ ID NO: 13, and an LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3. Further to this embodiment, the present invention contemplates use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. In one embodiment, the monoclonal antibodies or antibody fragments thereof provided herein can be used in a method for reducing inflammation in a mammal, as described in U.S. Patent No. 8,685,400, the contents of which are incorporated herein by reference in their entirety. The inflammation can be in the lungs and / or CNS. The inflammation in the lungs and / or CNS can be the result of a viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or affliction of the CNS, or a disease, condition, or affliction affecting the CNS. As provided herein, the disease, condition, or affliction of the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce innate immune or inflammasome-associated inflammation in a patient.The reduction may be compared to a control (e.g., an untreated patient and / or a patient before treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat pneumonia associated with a viral infection by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from viral-associated pneumonia, including ARDS and / or ALI. The monoclonal antibody or antibody fragment thereof of this embodiment may be present in a composition, such as a pharmaceutical composition as provided herein. In some cases, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agents may be any of the agents provided herein (e.g., EV uptake inhibitors) and / or antibodies or antibody fragments against other inflammasome components (e.g., IL-18, caspase-1, NLRP1, AIM2, etc.). Optionally, the other agents are hydroxychloroquine, chloroquine, and immunosuppressants, including, but not limited to, steroids, selective cytokine blockers (e.g., anakinra or tocilizumab), JAK inhibitors, interleukin inhibitors such as IL-1β and IL-6 inhibitors, TNF inhibitors, and CSF inhibitors, as well as anticoagulants such as low molecular weight heparin, tissue plasminogen activator, or enoxaparin. In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. The monoclonal antibody or antibody fragment thereof of this embodiment can be present in a composition, such as, for example, a pharmaceutical composition as provided herein. Optionally, the monoclonal antibody or fragment thereof is used in combination with one or more other agents in the treatment methods provided herein. The other agent may be any agent provided herein (e.g., an EV uptake inhibitor) and / or an antibody or antibody fragment against other inflammasome components (e.g., IL-18, caspase-1, NALP1, AIM2, etc.).

[0057] Provided herein are compositions and methods for reducing innate immune or inflammasome-associated inflammation. In some cases, the inflammasome-associated inflammation is in the CNS of a mammal suffering from or afflicted with a condition that results in or causes innate immune or inflammasome-associated inflammation. The compositions and methods described herein can include an antibody or active fragment thereof provided herein that specifically binds to at least one component of a mammalian inflammasome (e.g., ASC) and / or a compound that modulates (e.g., inhibits or reduces) extracellular vesicle (EV) uptake, and have utility as a treatment for CNS inflammation in a mammal. Examples of conditions that can result in inflammation in the CNS include viral infection, CNS injury (e.g., spinal cord injury (SCI), traumatic brain injury (TBI), or stroke), neurodegenerative disease, autoimmune disease (e.g., MS), asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis, interstitial lung disease, or acute respiratory distress syndrome. The composition can be administered in a therapeutically effective amount. The therapeutically effective amount can be a dose as provided herein. The agent may be an extracellular vesicle (EV) uptake inhibitor and / or an antibody or an active fragment thereof described herein that binds to a component of inflammasome, or a combination thereof. The composition may be administered by any suitable route, for example, by inhalation, intravenous, intraperitoneal, intranasal, or intracerebroventricular administration. The composition may further comprise at least one pharmaceutically acceptable carrier or diluent.

[0058] Provided herein are compositions and methods for treating viral infection or pulmonary inflammation, including ARDS, in a subject suffering from or suspected of suffering from a viral infection. The methods for treating virus-associated pulmonary inflammation provided herein can involve administering a composition (e.g., a pharmaceutical composition) containing an agent to a subject suffering from or suspected of suffering from pulmonary inflammation. The subject can exhibit clinical symptoms consistent with a viral infection. The subject can be diagnosed with any type of virus known in the art. The virus can be a coronavirus, such as SARS-CoV-2 or MERS coronavirus, a herpes virus, such as herpes simplex virus (HSV) and cytomegalovirus (CMV), or an influenza virus, such as influenza A H5N1 (avian influenza) and influenza A H1N1 (swine influenza).

[0059] Provided herein are compositions and methods for treating multiple sclerosis (MS) in a subject suffering from or suspected of suffering from MS. The methods for treating MS provided herein can involve administering a composition (e.g., a pharmaceutical composition) containing an agent to a subject suffering from or suspected of suffering from MS. Multiple sclerosis (MS) is an autoimmune disease that affects the brain and spinal cord. The subject can exhibit clinical symptoms consistent with MS. The subject can be diagnosed with any type of MS known in the art. The MS can be relapsing-remitting MS (RRMS), secondary progressive MS (SPMS), primary progressive MS (PPMS), or progressive relapsing MS (PRMS). The MS diagnosis can be or has been determined using any method known in the art. In one embodiment, the subject has been diagnosed with MS using the methods detailed in U.S. Patent No. 62 / 560,963, filed September 20, 2017, the contents of which are incorporated herein by reference in their entirety.

[0060] The agent may be a standard of care known in the art for MS or viral infection, an EV uptake inhibitor (e.g., any EV uptake inhibitor from Table 1), an antibody or active fragment thereof described herein (e.g., an anti-ASC monoclonal antibody or antibody fragment thereof) that binds to a component of the inflammasome, or any combination thereof. The composition may be administered by any appropriate route, for example, by inhalation, intravenous, intraperitoneal, intranasal, or intracerebroventricular administration. The composition may further comprise at least one pharmaceutically acceptable carrier or diluent. The standard of care may be selected from therapies aimed at modifying disease outcome, managing relapses, managing symptoms, or any combination thereof. Therapies aimed at modifying disease outcome may be selected from beta interferon, glatiramer acetate, fingolimod, teriflunomide, dimethyl fumarate, mitoxantrone, ocrelizumab, alemtuzumab, daclizumab, and natalizumab.

[0061] Provided herein are compositions and methods for reducing pulmonary inflammation in a mammal suffering from or afflicted with a condition that leads to or causes pulmonary inflammation. The compositions and methods described herein can include an antibody or active fragment thereof described herein that specifically binds to at least one component of a mammalian inflammasome (e.g., ASC), and / or a compound that modulates (e.g., inhibits or reduces) extracellular vesicle (EV) uptake, and have use as a treatment for pulmonary inflammation in a mammal.

[0062] Described herein are methods for reducing pulmonary inflammation in a mammal having a condition that leads to and / or causes a pulmonary inflammatory response. In one embodiment, the method for treating pulmonary inflammation in a mammal comprises administering to the mammal a composition comprising an agent that inhibits inflammasome signaling. The mammal may be a patient or subject described herein. Examples of conditions that may lead to pulmonary inflammation include central nervous system (CNS) injury (e.g., spinal cord injury (SCI), traumatic brain injury (TBI), or stroke), neurodegenerative disease, autoimmune disease (e.g., MS), asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis, interstitial lung disease, or acute respiratory distress syndrome. The composition can be administered in a therapeutically effective amount. The therapeutically effective amount can be a dose as provided herein. The agent can be an extracellular vesicle (EV) uptake inhibitor and / or an antibody or active fragment thereof described herein that binds to a component of the inflammasome, or a combination thereof. The composition can be administered by any suitable route, for example, by inhalation, intravenous, intraperitoneal, nasal, or intracerebroventricular. The composition may further comprise at least one pharmaceutically acceptable carrier or diluent.

[0063] In one embodiment, administration of an agent (e.g., an antibody or antibody fragment derived therefrom, alone or in combination with, e.g., an EV uptake inhibitor) in the methods provided herein can result in a decrease in the activity and / or expression level of a mammalian inflammasome component in the CNS or lung of a subject. The decrease can occur, for example, in lung cells, such as type II pneumocytes. The decrease can be compared to a control. The control can be the subject before administration of the agent. The control can also be the activity and / or expression level of an inflammasome component in a subject not administered the agent. In one embodiment, administration of the agent results in a decrease in caspase-1 activation in at least the CNS or CNS cells of the subject. In one embodiment, administration of the agent results in a decrease in caspase-1 activation in at least the lung or lung cells of the subject. In one embodiment, administration of the agent results in a decrease in the expression level of one or more inflammasome components (e.g., ASC, AIM2, NALP1, NALP2, NALP3, or NLRC4) in at least the CNS or CNS cells of the subject. In one embodiment, administration of the agent results in a decrease in the expression level of one or more inflammasome components (e.g., ASC, AIM2, NALP1, NALP2, NALP3, or NLRC4) in at least the lungs or lung cells of the subject.

[0064] In another embodiment, administration of an agent (e.g., an antibody or antibody fragment derived therefrom, alone or in combination with, e.g., an EV uptake inhibitor) can result in the reduction or elimination of acute lung injury (ALI). In one embodiment, the reduction in ALI is evidenced by a reduction in neutrophil infiltration into the alveoli and / or interstitial spaces, a reduction or elimination of alveolar septal thickening, or a combination thereof. The reduction may be compared to a control. The control may be ALI in a subject before administration of the agent. The control may be ALI in a subject with ALI who has not been administered the agent.

[0065] In yet another embodiment, administration of an agent (e.g., an antibody or antibody fragment derived therefrom, alone or in combination with, for example, an EV uptake inhibitor) can result in the reduction or elimination of pyroptosis in the CNS or lungs of a subject. Pyroptosis is an inflammation-induced form of cell death involving activation of caspase-1. Pyroptosis can be induced by caspase-1-mediated cleavage of gasdermin D (GSDMD). In one embodiment, the reduction of pyroptosis is evidenced by reduced or absent cleavage of GSDMD in the lungs or lung cells (e.g., type II pneumocytes) of the subject. The reduction or elimination of pyroptosis can be compared to a control. The reduction or absence of GSDMD cleavage can be compared to a control. The control can be the level of pyroptosis in the subject before administration of the agent. The control can be the level of pyroptosis in a subject with pyroptosis who has not been administered the agent.

[0066] The success of or response to the therapeutic methods provided herein (e.g., treatment of MS, innate immune or inflammasome-associated inflammation, CNS inflammation, and / or pulmonary inflammation) can be monitored by measuring the level of at least one inflammasome protein. Thus, in some embodiments, the therapeutic methods provided herein further include measuring the level of at least one inflammasome protein in a biological sample obtained from the subject after treatment, creating a therapeutic inflammasome protein signature associated with a positive response to treatment, wherein the therapeutic protein signature comprises a low level of at least one inflammasome protein, and identifying subjects exhibiting the presence of the therapeutic protein signature as having a positive response to treatment. A decrease in the level, abundance, or concentration of one or more inflammasome proteins (e.g., ASC, IL-18, or caspase-1) indicates the effectiveness of the treatment in the subject. The one or more inflammasome proteins measured in the sample obtained after treatment may be the same as or different from the inflammasome proteins measured in the sample obtained before treatment. The level of the inflammasome protein can also be used to adjust the dose or frequency of treatment. Inflammasome protein levels can be confirmed using the methods and techniques provided herein or as described in U.S. Patent Publication No. WO 2019 / 060516, filed September 20, 2018.

[0067] In one embodiment, the agent administered in the treatment methods provided herein is an EV uptake inhibitor, which may be a compound provided herein, an antisense RNA, an siRNA, a peptide, an antibody, or an active fragment thereof, or a combination thereof. The compound or peptide can be one or more compounds selected from heparin, α-difluoromethylornithine (DFMO), enoxaparin, asialofetuin, human receptor-associated protein (RAP), RGD (Arg-Gly-Asp) peptide, cytochalasin D, cytochalasin B, ethylenediaminetetraacetic acid (EDTA), latrunculin A, latrunculin B, NSC23766, Dynasoa, chlorpromazine, 5-(N-ethyl-N-isopropyl) amiloride (EIPA), amiloride, bafilomycin A, monensin, and chloroquine, annexin-V, wortmannin, LY294002, methyl-β-cyclodextrin (MPCD), filipin, simvastatin, fumonisin B1, and N-butyldeoxynojirimycin hydrochloride, U0126, or a proton pump inhibitor. The EV uptake inhibitor antibodies or active fragments thereof provided herein may be one or more antibodies or active fragments thereof against the protein targets set forth in Table 1. The composition for treating and / or reducing inflammation in the CNS or lungs of a mammal using an EV uptake inhibitor may further comprise at least one pharmaceutically acceptable carrier or diluent.

[0068] [Table 1]

[0069] [Table 2]

[0070] In one embodiment, the administered agent is an antibody or active fragment thereof described herein directed against a component of a mammalian inflammasome or an antigen or epitope derived therefrom. In another embodiment, the administered agent is an antisense RNA or siRNA directed against a component of a mammalian inflammasome. The inflammasome component can be any inflammasome component known in the art, such as, for example, NAPL1, NALP2, NALP3, NLRC4, or AIM2 inflammasome. In typical embodiments, the antibody specifically binds to ASC or an antigen or epitope derived therefrom. However, antibodies directed against other components of a mammalian inflammasome (e.g., NALP1, NALP2, NALP3, NLRC4, or AIM2 inflammasome) may also be used.

[0071] The antibodies described herein may be monoclonal or polyclonal antibodies, or active fragments thereof. The antibodies or active fragments may be chimeric, human, or humanized, as described herein.

[0072] Any suitable antibody or active fragment thereof described herein that specifically binds to ASC can be used, for example, an antibody that inhibits ASC activity in the CNS (e.g., CNS cells) or lung cells (e.g., type II alveolar cells) of a subject can be used. In one embodiment, the antibody specifically binds to an amino acid sequence having at least 85% sequence identity to the amino acid sequence SEQ ID NO: 1 or SEQ ID NO: 2. In another embodiment, the antibody or fragment thereof binds to an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence KKFKLKLLSVPLREGYGRIPR (SEQ ID NO: 5). In yet another embodiment, the antibody or fragment thereof binds to the amino acid sequence KKFKLKLLSVPLREGYGRIPR (SEQ ID NO: 5), or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids of SEQ ID NO: 5. In yet another embodiment, the antibody or fragment thereof binds to 2-5, 5-10, 10-15, or 15-20 amino acids of SEQ ID NO: 5. In some embodiments, the epitope of ASC bound by the antibody or antibody fragment (e.g., the epitope having amino acid SEQ ID NO: 5) is contiguous. In some embodiments, the epitope of ASC bound by the antibody or antibody fragment (e.g., the epitope having amino acid SEQ ID NO: 5) is discontinuous. In some instances, the monoclonal antibodies or antibody fragments thereof provided herein inhibit or reduce the activity of ASC.

[0073] As used herein, the term "epitope" includes any protein determinant capable of specific binding to an immunoglobulin or immunoglobulin fragment. Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three-dimensional structural characteristics, as well as specific charge characteristics. The term "epitope" also refers to the unit of structure typically bound by a pair of immunoglobulin heavy chain variable (VH) and light chain variable (VL) regions. An epitope defines the minimal binding site of an antibody and represents the target of antibody specificity.

[0074] Similarly, in another embodiment, the inflammasome is an NALP1 inflammasome, and at least one component is NALP1 (i.e., NLRP1). In this embodiment, the antibodies or active fragments thereof provided herein specifically bind to an amino acid sequence having at least 85% sequence identity to the amino acid sequence SEQ ID NO:3 or SEQ ID NO:4.

[0075] In yet another embodiment, the agent is one or more EV uptake inhibitors in combination with one or more antibodies or active fragments thereof described herein that bind to a component of an inflammasome. The EV uptake inhibitor may be any EV uptake inhibitor provided herein. The antibody that binds to a component of an inflammasome may be any antibody that binds to any inflammasome component provided herein. In one embodiment, the agent administered to a subject suffering from CNS or lung inflammation comprises a combination of heparin (e.g., enoxaparin) and an antibody that binds to a component of the AIM2 inflammasome (e.g., ASC).

[0076] In one embodiment, the method comprises obtaining a therapeutically effective amount of a composition comprising an antibody or active fragment thereof described herein that specifically binds to at least one component (e.g., ASC) of a mammalian inflammasome (e.g., AIM2 inflammasome); and administering the composition to a mammal suffering from CNS or lung inflammation, viral infection, or MS, wherein administering the composition to the mammal results in reduced caspase-1 activation in the CNS or lung of the mammal. In another embodiment, the method comprises obtaining a therapeutically effective amount of a composition comprising an antibody that specifically binds to at least one component (e.g., ASC) of a mammalian inflammasome (e.g., AIM2 inflammasome); and administering the composition to a mammal suffering from CNS or lung inflammation or MS, wherein administering the composition to the mammal reduces the level of one or more inflammasome components (e.g., ASC). In yet another embodiment, the method comprises obtaining a therapeutically effective amount of a composition comprising an antibody that specifically binds to at least one component (e.g., ASC) of a mammalian inflammasome (e.g., AIM2 inflammasome); and administering the composition to a mammal suffering from CNS or pulmonary inflammation or MS, wherein administering the composition to the mammal results in a reduction of ALI. The CNS or pulmonary inflammation can be due to CNS injury (e.g., SCI or TBI), asthma, chronic obstructive pulmonary disease (COPD), neurodegenerative disease, or an autoimmune disease with an inflammatory component. In one embodiment, the pulmonary inflammation is caused by a CNS injury, such as TBI or SCI. In another embodiment, the pulmonary inflammation occurs as a result of a viral infection, such as infection with a coronavirus or influenza virus.

[0077] In one embodiment, the method provided herein further involves detecting the level or activity of one or more components of a mammalian inflammasome in a sample from a subject suspected of suffering from CNS or pulmonary inflammation or MS. The method for detecting the level or activity involves measuring the level of at least one inflammasome protein (e.g., ASC or AIM2) in a sample obtained from the subject; and determining whether or not the level or activity of the at least one inflammasome protein (e.g., ASC or AIM2) is elevated. The level or activity of the at least one inflammasome protein can be elevated relative to the level of the at least one inflammasome protein in a control sample. The level or activity of the at least one inflammasome protein in the protein signature can be elevated relative to a predetermined baseline value or range of baseline values. The at least one inflammasome protein can be nucleotide-binding leucine-rich repeat pyrin domain-containing protein 1 (NLRP1), NLRP2, NLRP3, NLRC4, AIM2, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase-1, or a combination thereof. The sample may be cerebrospinal fluid (CSF), saliva, blood, serum, plasma, urine or lung aspirate.

[0078] An antibody that specifically binds to at least one component of a mammalian inflammasome. The methods described herein for reducing inflammation in the CNS and / or lungs of a mammal include compositions containing an antibody or active fragment thereof described herein that specifically binds to at least one component (e.g., ASC, AIM2) of a mammalian inflammasome (e.g., the AIM2 inflammasome). Compositions for treating and / or reducing inflammation in the CNS and / or lungs of a mammal may further comprise at least one pharmaceutically acceptable carrier or diluent. Exemplary antibodies against components of a mammalian inflammasome for use in the methods described herein may be those found in U.S. Pat. No. 8,685,400, the contents of which are incorporated herein by reference in their entirety. Also provided herein are exemplary monoclonal antibodies or antibody fragments, such as a monoclonal antibody or antibody fragment comprising a VH region having a VH region amino acid sequence comprising an HCDR1 of SEQ ID NO: 6, an HCDR2 of SEQ ID NO: 7, and an HCDR3 of SEQ ID NO: 8, and a VL region having a VL region amino acid sequence comprising an LCDR1 of SEQ ID NO: 12, an LCDR2 of SEQ ID NO: 13, and an LCDR3 of SEQ ID NO: 14.

[0079] In one embodiment, the composition for treating and / or reducing inflammation in the CNS or lungs of a mammal comprises an antibody or an active fragment thereof described herein that specifically binds to a domain or a portion thereof of a mammalian ASC protein, such as, for example, a human, mouse, or rat ASC protein. Any suitable anti-ASC antibody can be used, and several antibodies are commercially available. Examples of anti-ASC antibodies used in the methods herein include those found in U.S. Patent No. 8,685,400 (the contents of which are incorporated herein by reference in their entirety). Examples of commercially available anti-ASC antibodies for use in the methods described herein include, but are not limited to, 04-147 anti-ASC from MilliporeSigma, clone 2EI-7 mouse monoclonal antibody, AB3607-anti-ASC antibody from MilliporeSigma, orb194021 anti-ASC from Biorbyt, LS-C331318-50 anti-ASC from LifeSpan Biosciences, AF3805 anti-ASC from R&D Systems, NBP1-78977 anti-ASC from Novus Biologicals, 600-401-Y67 anti-ASC from Rockland Immunochemicals, D086-3 anti-ASC from MBL International, AL177 anti-ASC from Adipogen, monoclonal anti-ASC (clone o93E9) antibody from Santa Cruz Biotechnology, anti-ASC antibody (F-9), anti-ASC antibody (B-3) from Santa Cruz Biotechnology, Enzo Life ASC polyclonal antibody - ADI-905-173 from Biosciences, or A161 anti-human ASC - Leinco Technologies. Human ASC protein can be under accession numbers NP_037390.2 (Q9ULZ3-1), NP_660183 (Q9ULZ3-2), or Q9ULZ3-3. Rat ASC protein can be under accession number NP_758825 (BAC43754). Mouse ASC protein can be under accession number NP_075747.3.In one embodiment, the antibody binds to the PYRIN-PAAD-DAPIN domain (PYD) of a mammalian ASC protein (e.g., human, mouse, or rat ASC), or a portion or fragment thereof. In this embodiment, the antibody described herein specifically binds to an amino acid sequence having at least 65% (e.g., 65, 70, 75, 80, 85%) sequence identity to the PYD domain of human, mouse, or rat ASC, or a fragment thereof. In one embodiment, the antibody binds to the C-terminal caspase recruitment domain (CARD) of a mammalian ASC protein (e.g., human, mouse, or rat ASC), or a portion or fragment thereof. In this embodiment, the antibody described herein specifically binds to an amino acid sequence having at least 65% (e.g., 65, 70, 75, 80, 85%) sequence identity to the CARD domain of human, mouse, or rat ASC, or a fragment thereof. In yet another embodiment, the antibody binds to a portion or fragment thereof of a mammalian ASC protein sequence (e.g., human, mouse, or rat ASC) located between the PYD domain and the CARD domain. In another embodiment, a composition for treating and / or reducing inflammation in the CNS and / or lungs of a mammal comprises an antibody that specifically binds to a region of rat ASC, e.g., the amino acid sequence ALRQTQPYLVTDLEQS (SEQ ID NO: 1) (i.e., residues 178-193 of rat ASC, Accession No. BAC43754). In this embodiment, the antibody described herein specifically binds to an amino acid sequence having at least 65% (e.g., 65, 70, 75, 80, 85%) sequence identity to the amino acid sequence ALRQTQPYLVTDLEQS (SEQ ID NO: 1) of rat ASC. In another embodiment, a composition for treating and / or reducing inflammation in the CNS and / or lungs of a mammal comprises an antibody that specifically binds to a region of human ASC, e.g., the amino acid sequence RESQSYLVEDLERS (SEQ ID NO: 2).In yet another embodiment, a composition for treating and / or alleviating inflammation in the CNS and / or lungs of a mammal comprises an antibody that specifically binds to a region of human ASC, e.g., the amino acid sequence KKFKLKLLSVPLREGYGRIPR (SEQ ID NO: 5; i.e., residues 21-41 of human ASC), or amino acids 5-10, 10-15, or 15-20 of SEQ ID NO: 5. In one embodiment, an antibody that binds to an ASC domain described herein or a fragment thereof inhibits ASC activity in mammalian lung cells, e.g., type II pneumocytes. In another embodiment, an antibody that binds to an ASC domain described herein or a fragment thereof (e.g., a monoclonal anti-ASC antibody or antibody fragment thereof provided herein) inhibits ASC activity in the CNS of a mammal suffering from or suspected of suffering from a CNS injury or disorder. Examples of CNS injuries or disorders include TBI, SCI, stroke, amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction, muscular CNS weakness, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD).

[0080] In certain embodiments, the present invention provides antibodies and antibody fragments that specifically bind to ASC and comprise one or more amino acid sequences set forth in Table 2. Also provided herein are isolated nucleic acid molecules encoding monoclonal antibodies or antibody fragments thereof comprising a nucleic acid sequence set forth in Table 2. Optionally, expression vectors comprise the nucleic acid molecules of Table 2. The expression vectors may comprise heavy or light chain constant regions. An example of a light and heavy chain expression vector system for use in the compositions and methods provided herein is the Antitope pANT expression vector system for IgG4(S241P) heavy chain and κ light chain. The heavy or light chain nucleic acid molecules can be operably linked to suitable regulatory sequences for expression of the nucleic acid segment in a host cell.

[0081] [Table 3]

[0082]

Table 4

[0083]

Table 5

[0084]

Table 6

[0085]

Table 7

[0086] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, and the amino acid sequence of the VH region comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lungs and / or CNS. The inflammation in the lungs and / or CNS may be due to a viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or disorder of or affecting the CNS. As provided herein, diseases, conditions, or afflictions of or affecting the CNS can include stroke, autoimmune diseases and / or CNS disorders such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction-induced CNS weakness, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of a monoclonal antibody or antibody fragment thereof in a method for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. This reduction may be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, a monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition as provided herein.

[0087] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VL region comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30, or 31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lungs and / or CNS. The inflammation in the lungs and / or CNS may be due to a viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or disorder of or affecting the CNS. As provided herein, diseases, conditions, or afflictions of or affecting the CNS can include stroke, autoimmune diseases and / or CNS disorders such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction-induced CNS weakness, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of a monoclonal antibody or antibody fragment thereof in a method for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. This reduction may be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, a monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition as provided herein.

[0088] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22; and the amino acid sequence of the VL region comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30, or 31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation can be innate immune inflammation. The inflammation can be inflammasome-associated inflammation. The inflammation can occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition.The composition can be a pharmaceutical composition provided herein.

[0089] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 18, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the amino acid sequence of the VL region comprises SEQ ID NO: 28, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0090] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 18, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the amino acid sequence of the VL region comprises SEQ ID NO: 29, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 29. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0091] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, the antibody or antibody fragment thereof comprising a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 18, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the amino acid sequence of the VL region comprises SEQ ID NO: 30, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 30. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation can be innate immune inflammation. The inflammation can be inflammasome-associated inflammation. The inflammation can occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0092] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 18, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the amino acid sequence of the VL region comprises SEQ ID NO: 31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0093] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 19, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the amino acid sequence of the VL region comprises SEQ ID NO: 28, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0094] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 19 or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the amino acid sequence of the VL region comprises SEQ ID NO: 29 or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 29. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0095] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 19 or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the amino acid sequence of the VL region comprises SEQ ID NO: 30 or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 30. In some cases, a monoclonal antibody or antibody fragment derived therefrom comprising a VH region amino acid sequence comprising SEQ ID NO: 19 and a VL region amino acid sequence comprising SEQ ID NO: 30 can be referred to as IC-100. In addition to this embodiment, provided herein is the use of a monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation can be innate immune inflammation. The inflammation can be inflammasome-associated inflammation. The inflammation can occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS.In some cases, the monoclonal antibody or antibody fragment thereof of the present embodiments is present in a composition, which can be a pharmaceutical composition as provided herein.

[0096] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 19, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the amino acid sequence of the VL region comprises SEQ ID NO: 31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition as provided herein.

[0097] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:20, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:20; and the amino acid sequence of the VL region comprises SEQ ID NO:28, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:28. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0098] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:20, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:20; and the amino acid sequence of the VL region comprises SEQ ID NO:29, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:29. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0099] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:20, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:20; and the amino acid sequence of the VL region comprises SEQ ID NO:30, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:30. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition as provided herein.

[0100] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:20, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:20; and the amino acid sequence of the VL region comprises SEQ ID NO:31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0101] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:21, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:21; and the amino acid sequence of the VL region comprises SEQ ID NO:28, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:28. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0102] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:21, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:21; and the amino acid sequence of the VL region comprises SEQ ID NO:29, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:29. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0103] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:21, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:21; and the amino acid sequence of the VL region comprises SEQ ID NO:30, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:30. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0104] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:21, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:21; and the amino acid sequence of the VL region comprises SEQ ID NO:31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0105] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the amino acid sequence of the VL region comprises SEQ ID NO: 28, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0106] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the amino acid sequence of the VL region comprises SEQ ID NO: 29, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 29. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. The inflammation may occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can result from viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition as provided herein.

[0107] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO: 22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the amino acid sequence of the VL region comprises SEQ ID NO: 30, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 30. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation can be innate immune inflammation. The inflammation can be inflammasome-associated inflammation. The inflammation can occur in the lung and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment). In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0108] In one embodiment, provided herein is a monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain or kappa chain variable (VL) region, wherein the amino acid sequence of the VH region comprises SEQ ID NO:22, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:22; and the amino acid sequence of the VL region comprises SEQ ID NO:31, or an amino acid sequence at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:31. In addition to this embodiment, provided herein is the use of the monoclonal antibody or antibody fragment thereof in a method for treating inflammation in a subject. The inflammation may be innate immune inflammation. The inflammation may be inflammasome-associated inflammation. In one embodiment, the monoclonal antibody or antibody fragment thereof provided herein can be used in a method for reducing inflammation in a mammal, as described in U.S. Pat. No. 8,685,400, the contents of which are incorporated herein by reference in their entirety. Inflammation can occur in the lungs and / or CNS. Inflammation in the lungs and / or CNS can be due to viral infection, injury (e.g., traumatic brain injury (TBI) or spinal cord injury (SCI)), or a disease, condition, or morbidity of or affecting the CNS. As provided herein, the disease, condition, or morbidity of or affecting the CNS can be stroke, as well as autoimmune and / or CNS diseases such as amyotrophic lateral sclerosis (ALS), Lou Gehrig's disease, multiple sclerosis (MS), immune dysfunction muscle CNS wasting, muscular dystrophy (MD), Alzheimer's disease (AD), and Parkinson's disease (PD). The use of monoclonal antibodies or antibody fragments thereof in methods for treating inflammation can reduce inflammation in the CNS and / or lungs of a patient. The reduction can be compared to a control (e.g., an untreated patient and / or a patient prior to treatment).In one embodiment, the monoclonal antibody or antibody fragment derived therefrom is used to treat MS by administering the monoclonal antibody or antibody fragment derived therefrom to a patient suffering from or suspected of suffering from MS. Optionally, the monoclonal antibody or antibody fragment thereof of this embodiment is present in a composition. The composition may be a pharmaceutical composition provided herein.

[0109] In another embodiment, the composition for reducing inflammation in the CNS or lungs of a mammal comprises an antibody or an active fragment thereof described herein that specifically binds to NLRP1 (e.g., an anti-NLRP1 chicken antibody) or a domain thereof. Any suitable anti-NLRP1 antibody can be used, and some antibodies are commercially available. Examples of anti-NLRP1 antibodies used in the methods herein include those described in U.S. Patent No. 8,685,400 (the contents of which are incorporated herein by reference in their entirety).Examples of commercially available anti-NLRP1 antibodies for use in the methods described herein include, but are not limited to, human NLRP1 polyclonal antibody AF6788 from R&D Systems, rabbit polyclonal anti-NLRP1 ABF22 from EMD Millipore, rabbit polyclonal anti-NLRP1 NB100-56148 from Novus Biologicals, mouse polyclonal anti-NLRP1 SAB1407151 from Sigma-Aldrich, rabbit polyclonal anti-NLRP1 ab3683 from Abcam, rabbit polyclonal anti-NLRP1 orb325922 from Biorbyt, rabbit polyclonal anti-NLRP1 MBS7001225 from mybiosource, sheep polyclonal AF6788 from R&D systems, mouse monoclonal anti-NLRP1 oaed00344 from Aviva Systems, and rabbit polyclonal anti-NLRP1 from Aviva Systems. ARO54478_P050, rabbit polyclonal anti-NLRP1 APO7775PU-N from Origene, rabbit polyclonal anti-NLRP1 ABIN768983 from Antibodies online, rabbit polyclonal anti-NLRP1 3037 from Prosci, rabbit polyclonal anti-NLRP1 12256-1-AP from Proteintech, mouse monoclonal anti-NLRP1 ALX-804-803-C100 from Enzo, mouse monoclonal anti-NLRP1 MA1-25842 from Invitrogen, mouse monoclonal anti-NLRP1 GTX16091 from GeneTex, rabbit polyclonal anti-NLRP1 200-401-CX5 from Rockland, or rabbit polyclonal anti-NLRP1 4990 from Cell Signaling Technology. The human NLRP1 protein may be accession number AAH51787, NP_001028225, NP_055737, NP_127497, NP_127499, or NP_127500. In one embodiment, the antibody binds to the Pyrin, NACHT, LRR1-6, FUND, or CARD domain of a mammalian NLRP1 protein (e.g., human NLRP1), or a portion or fragment thereof.In this embodiment, the antibodies described herein specifically bind to an amino acid sequence having at least 65% (e.g., 65, 70, 75, 80, 85%) sequence identity with a particular domain of human NLRP1 (e.g., Pyrin, NACHT, LRR1-6, FUND, or CARD) or a fragment thereof. In one embodiment, a chicken anti-NLRP1 polyclonal antibody custom designed and manufactured by Ayes Laboratories is used to reduce lung inflammation. The antibody may be directed against the following amino acid sequence of human NLRP1: CEYYTEIREREREKSEKGR (SEQ ID NO: 3). In one embodiment, an antibody that binds to the NLRP1 domain or a fragment thereof described herein inhibits NLRP1 activity in lung cells, e.g., mammalian type II pneumocytes.

[0110] In yet another embodiment, the composition for reducing inflammation in the CNS or lungs of a mammal comprises the antibody or its active fragment described herein, which specifically binds to AIM2 or its domain.Any suitable anti-AIM2 antibody can be used, and some antibodies are commercially available. Examples of commercially available anti-AIM2 antibodies for use in the methods provided herein include, but are not limited to, the following: rabbit polyclonal anti-AIM2 from Proteintech (catalog number 20590-1-AP), Abcam anti-AIMS antibody (ab119791), rabbit polyclonal anti-AIM2 (N-terminal region) from ECM biosciences (catalog number AP3851), rabbit polyclonal anti-ASC from Elabsciences (catalog number E-AB-30449), anti-AIM2 mouse monoclonal antibody designated AIM2 Antibody (3C4G11) from Santa Cruz Biotechnology (catalog number sc-293174), mouse monoclonal AIM2 antibody from Origene with catalog number TA324972, ThermoFisher AIM2 monoclonal antibody (10M2B3) from Scientific, AIM2 rabbit polyclonal antibody ABIN928372 or ABIN760766 from Antibodies-online, Biomatix coated anti-AIM2 polyclonal antibody with catalog number CAE02153, anti-AIM2 polyclonal antibody (OABF01632) from Aviva Systems Biology, rabbit polyclonal anti-AIM2 antibody LS-C354127 from LSBio, rabbit monoclonal anti-AIM2 antibody (catalog number MA5-16259) from Cell Signaling Technology, and rabbit polyclonal anti-AIM2 monoclonal antibody (catalog number AIM2) from Fab Gennix International Incorporated. 201AP, MyBiosource rabbit polyclonal anti-AIM2 (catalog no. MBS855320), Signalway rabbit polyclonal anti-AIM2 (catalog no. 36253), NovusRabbit polyclonal anti-AIM2 from Biologics (catalog number 43900002), rabbit polyclonal anti-AIM2 GTX54910 from GeneTex, rabbit polyclonal anti-AIM2 26-540 from Prosci, mouse monoclonal anti-AIM2 orb333902 from Biorbyt, rabbit polyclonal anti-AIM2 ab93015 from Abcam, rabbit polyclonal anti-AIM2 ab76423 from Abcam, mouse polyclonal anti-AIM2 SAB1406827 from Sigma-Aldrich, or Biolegend anti-AIM2 3B10. Human AIM2 protein can be under the accession numbers NX_014862, NP004824, XP016858337, XP005245673, AAB81613, BAF84731, or AAH10940. In one embodiment, the antibody binds to the Pyrin or HIN-200 domain of a mammalian AIM2 protein (e.g., human AIM2) or a part or fragment thereof. In this embodiment, the antibody described herein specifically binds to an amino acid sequence having at least 65% (e.g., 65, 70, 75, 80, 85%) sequence identity with a specific domain of human AIM2 (e.g., Pyrin or HIN-200) or a fragment thereof. In one embodiment, the antibody that binds to the AIM2 domain or a fragment thereof described herein inhibits AIM2 activity in lung cells, such as mammalian type II pneumocytes.

[0111] Anti-inflammasome (e.g., anti-ASC, anti-NLRP1, or anti-AIM2) antibodies described herein include polyclonal and monoclonal rodent antibodies, polyclonal and monoclonal human antibodies, or any portion thereof, having at least one antigen-binding region of an immunoglobulin variable region, which specifically bind to a component of a mammalian inflammasome, such as ASC or AIM2 (e.g., the AIM2 inflammasome). In some cases, such antibodies are specific for ASC, such as an antibody being specific for ASC if it is raised against an epitope of a polypeptide and binds to at least a portion of a natural or recombinant protein.

[0112] In certain embodiments, the antibodies provided herein include polypeptides having one or more amino acid substitutions, deletions, or insertions. For example, anti-ASC monoclonal antibodies or ASC-binding antibody fragments include polypeptides having one or more amino acid substitutions, deletions, or insertions compared to polypeptides having one or more amino acid sequences of SEQ ID NOs: 6-8, 12-14, 18-22, or 28-31. The antibodies provided herein may have one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid substitutions, deletions, or insertions. For example, anti-ASC monoclonal antibodies or ASC-binding antibody fragments may have one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid substitutions, deletions, or insertions. Substitutions, deletions, or insertions can be introduced by standard techniques, such as site-directed mutagenesis or PCR-mediated mutagenesis, of nucleic acid molecules encoding the anti-ASC antibody or ASC-binding antibody fragment polypeptides.

[0113] In certain embodiments, conservative amino acid substitutions are made at one or more positions in the amino acid sequence of an antibody or antibody fragment disclosed herein. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. In certain embodiments, conservative amino acid substitutions are made only in the FR sequences of an antibody or antibody fragment, and not in the CDR sequences. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan; histidine). Thus, for example, an amino acid residue in an anti-ASC monoclonal antibody or ASC-binding antibody fragment polypeptide can be replaced with another amino acid residue from the same side chain family. In certain embodiments, a chain of amino acids can be replaced with a structurally similar chain that differs in the order and / or composition of the members of the side chain family. A person skilled in the art will be able to assess whether an anti-ASC monoclonal antibody or ASC-binding antibody fragment comprising a polypeptide having one or more amino acid substitutions, deletions or insertions compared to a polypeptide having one or more amino acid sequences of SEQ ID NOs: 6-8, 12-14, 18-22 or 28-31 binds to an ASC protein using routine, skilled artisan-recognized methods, including but not limited to, ELISA, Western blot, phage display, etc.

[0114] Calculations of sequence homology or identity between sequences (the terms are used interchangeably herein) can be performed as follows.

[0115] To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced into one or both of the first and second amino acid or nucleic acid sequences for optimal alignment, and non-homologous sequences can be ignored for comparison purposes). In exemplary embodiments, the length of the reference sequence aligned for comparison purposes is at least 30%, 40%, 50%, 60%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position (as used herein, amino acid or nucleic acid "identity" is equivalent to amino acid or nucleic acid "homology"). The percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps and the length of each gap that need to be introduced for optimal alignment of the two sequences.

[0116] Comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In one embodiment, percent identity between two amino acid sequences is determined using the Needleman et al. ((1970) J. Mol. Biol. 48:444-453) algorithm incorporated into the GAP program of the GCG software package (available at www.gcg.com), using either a BLOSUM 62 matrix or a PAM250 matrix, a gap weight of 16, 14, 12, 10, 8, 6, or 4, and a length weight of 1, 2, 3, 4, 5, or 6. In yet another embodiment, percent identity between two nucleotide sequences is determined using the GAP program of the GCG software package (available at www.gcg.com), using a NWSgapdna.CMP matrix, a gap weight of 40, 50, 60, 70, or 80, and a length weight of 1, 2, 3, 4, 5, or 6. One set of parameters (and parameters that can be used if the artisan is unsure which parameters to apply to determine whether a molecule falls within the sequence identity or homology limits of the invention) is the BLOSUM62 scoring matrix, with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

[0117] The percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of Meyers et al. ((1989) CABIOS 4:11-17) as incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0118] In certain embodiments, the antibody is a monoclonal antibody. In other embodiments, the antibody is a polyclonal antibody. The term "monoclonal antibody" refers to a population of antibody molecules that contain only one species of antigen-binding site capable of immunoreacting with a particular epitope of an antigen. Thus, a monoclonal antibody composition typically exhibits a single binding affinity for a particular protein with which it immunoreacts.

[0119] Optionally, the antibody (anti-ASC monoclonal antibody or ASC binding antibody fragment) of the present invention is a humanized, chimeric or human antibody.

[0120] In some embodiments, the antibodies of the invention are humanized antibodies.

[0121] "Humanized antibody," as the term is used herein, refers to an antibody that has been engineered to contain one or more human framework regions in the variable regions together with non-human (e.g., mouse, rat, or hamster) complementarity-determining regions (CDRs) of the heavy and / or light chains. In certain embodiments, a humanized antibody comprises sequences that are entirely human except for the CDR regions. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, a humanized antibody may comprise residues that are not found in the human form of the antibody or in the imported CDR or framework sequences, but are included to further refine and optimize antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. The FR regions can be modified by any method known in the art and / or provided herein. Modifications can confer desirable properties, such as increased half-life and / or improved expression in host cells. In one embodiment, the FR regions can be modified or mutated as described in U.S. Patent No. 20150232557, incorporated herein by reference. Other forms of humanized antibodies can have one or more CDRs (CDR L1, CDR L2, CDR L3, CDR H1, CDR H2, or CDR H3) that are altered with respect to the original antibody, also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody. Humanized antibodies optimally comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin.

[0122] Humanized antibodies generally have lower immunogenicity in humans than non-humanized antibodies, and therefore offer therapeutic advantages in certain situations. For example, antibody constant regions can be engineered to be immunologically inert (e.g., not induce complement lysis). See, for example, PCT Publication No. PCT / GB99 / 01441; UK Patent Application No. 9809951.8 (each of which is incorporated herein by reference in its entirety). Those skilled in the art will be familiar with humanized antibodies and suitable techniques for their production. See, for example, Hwang, WYK, et al., Methods 36:35, 2005; Queen et al., Proc. Natl. Acad. Sci. USA, 86:10029-10033, 1989; Jones et al., Nature, 321:522-25, 1986; Riechmann et al., Nature, 332:323-27, 1988; Verhoeyen et al., Science, 239:1534-36, 1988; Orlandi et al., Proc. Natl. Acad. Sci. USA, 86:3833-37, 1989; U.S. Patent Nos. 5,225,539; 5,530,101; 5,585,089; 5,693,761; 5,693,762; 6,180,370; and Selick et al., WO 90 / 07861 (each of which is incorporated by reference in its entirety). Other methods of humanizing antibodies that can be used are disclosed by Daugherty et al., Nucl. Acids Res. 19:2471-2476, 1991; and in U.S. Patent Nos. 6,180,377; 6,054,297; 5,997,867; 5,866,692; 6,210,671; and 6,350,861; and PCT Publication No. WO 01 / 27160, each of which is incorporated herein by reference in its entirety.For example, an anti-ASC antibody or anti-ASC antigen-binding fragment of the present invention comprises a VH region amino acid sequence comprising HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and HCDR3 of SEQ ID NO: 8; a VL region amino acid sequence comprising LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14; and one or more human framework region sequences.

[0123] In some embodiments, the antibody of the present invention is a chimeric antibody and specifically binds to ASC. In some cases, anti-ASC chimeric antibodies reduce the activity of ASC. As used herein, the term "chimeric antibody" refers to an antibody engineered to contain at least one human constant region. For example, one or all of the variable regions of the light chain and / or one or all of the variable regions of the heavy chain of a mouse antibody (e.g., a mouse monoclonal antibody) can each be linked to a human constant region, such as, but not limited to, an IgG1 human constant region. Chimeric antibodies are typically less immunogenic in humans than non-chimeric antibodies, and therefore offer therapeutic benefits in certain situations. Those skilled in the art will be familiar with chimeric antibodies and suitable techniques for their production. See, e.g., Cabilly et al., U.S. Pat. No. 4,816,567; Shoemaker et al., U.S. Pat. No. 4,978,775; Beavers et al., U.S. Pat. No. 4,975,369; and Boss et al., U.S. Pat. No. 4,816,397 (each of which is incorporated by reference herein in its entirety). For example, an antibody or antigen-binding fragment of the invention may comprise a VH region comprising SEQ ID NO: 22; a VL region comprising SEQ ID NO: 31; and a human constant region.

[0124] As used herein, the terms "immunological binding" and "immunological binding properties" refer to the type of non-covalent interactions that occur between an immunoglobulin molecule (e.g., an antibody) and an antigen for which the immunoglobulin is specific. The strength or affinity of an immunological binding interaction is determined by the dissociation constant (K d ) and K dA smaller K represents a higher affinity. The immunological binding properties of a selected polypeptide can be quantified using methods well known in the art. One such method involves measuring the rates of formation and dissociation of the antigen-binding site / antigen complex, which depend on the concentrations of the complex partners, the affinity of the interaction, and geometric parameters that affect the rates in both directions equally. Thus, the "on rate constant" (K on ) and the "off rate constant" (K off ) can be determined by calculating the concentration and the actual binding and dissociation rates (see Nature 361:186-87 (1993)). off / K on The ratio of α to β allows the release of all parameters unrelated to affinity and the dissociation constant K d (For a review, see Davies et al. (1990) Annual Rev Biochem 59:439-473). The equilibrium binding constant (K) as measured by an assay such as a radioligand binding assay or similar assay known to those skilled in the art is d An antibody of the present invention is said to specifically bind to an epitope (eg, an ASC fragment having amino acid sequence SEQ ID NO: 5) when the binding affinity of the antibody to the epitope is ≦10 μM, ≦10 nM, ≦10 nM, and ≦100 pM to about 1 pM.

[0125] In particular embodiments, the antibodies of the invention are monovalent or bivalent and comprise a single chain or two chains. Functionally, the binding affinity of the antibodies is 10 -5 M~10 -12 For example, the binding affinity of an antibody is in the range of 10 -6 M~10 -12 M, 10 -7 M~10 -12 M, 10 -8 M~10 -12 M, 10 -9 M~10 -12 M, 10 -5 M~10 -11 M, 10 -6 M~10 -11 M, 10 -7 M~10 -11 M, 10 -1M~10 -11 M, 10 -9 M~10 -11 M, 10 -10 M~10 -11 M, 10 -5 M~10 -10 M, 10 -6 M~10 -10 M, 10 -7 M~10 -10 M, 10 -8 M~10 -10 M, 10 -9 M~10 -10 M, 10 -5 M~10 -9 M, 10 -6 M~10 -9 M, 10 -7 M~10 -9 M, 10 -8 M~10 -9 M, 10 -5 M~10 -8 M, 10 -6 M~10 -8 M, 10 -7 M~10 -8 M, 10 -5 M~10 -7 M, 10 -6 M~10 -7 M or 10 -5 M~10 -6 I am M.

[0126] Methods for determining the specificity and affinity of monoclonal antibodies by competitive inhibition are described in Harlow, et al: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988; Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, NY, (1992, 1993); and Muller, Meth. Enzymol. 92:589-601, 1983, which are incorporated herein by reference in their entireties.

[0127] Anti-inflammasome (e.g., anti-ASC and anti-AIM2) antibodies of the present invention can be routinely produced according to methods such as, but not limited to, inoculation of appropriate animals with polypeptides or antigenic fragments, in vitro stimulation of lymphocyte populations, synthetic methods, hybridomas, and / or recombinant cells expressing nucleic acids encoding such anti-ASC or anti-NLR1 antibodies. Immunization of animals with purified recombinant ASC or a peptide fragment thereof, e.g., residues 178-193 (SEQ ID NO: 1) of rat ASC (e.g., accession number BAC43754), SEQ ID NO: 2 of human ASC, or residues 21-41 (SEQ ID NO: 5) of human ASC (e.g., accession number NP_037390.2), is an example of a method for preparing anti-ASC antibodies. Similarly, immunization of animals with purified recombinant NLRP1 or a peptide fragment thereof, e.g., residues MEE SQS KEE SNT EG-cys (SEQ ID NO: 4) of rat NALP1 or SEQ ID NO: 3 of human NALP1, is an example of a method for preparing anti-NLRP1 antibodies.

[0128] Monoclonal antibodies that specifically bind to ASC or NLRP1 can be obtained by methods well known to those skilled in the art. For example, see Kohler and Milstein, Nature 256:495-497, 1975; U.S. Patent No. 4,376,110; Ausubel et al., eds., Current Protocols in Molecular Biology, Greene Publishing Assoc. and Wiley Interscience, NY, (1987, 1992); Harlow and Lane, ANTIBODIES: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988; Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, NY, (1992, 1993) (the contents of which are incorporated herein by reference in their entirety). Such antibodies may be of any immunoglobulin class, including IgG, IgM, IgE, IgA, GILD, and any subclass thereof. Hybridomas producing the monoclonal antibodies of the present disclosure can be cultured in vitro, in situ, or in vivo. In one embodiment, the hybridoma producing the anti-ASC monoclonal antibody of the present disclosure is the ICCN1.OH hybridoma. In another embodiment, the hybridoma producing the anti-ASC monoclonal antibody of the present disclosure produces a monoclonal antibody comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the amino acid sequence of the VH region comprises an HCDR1 of SEQ ID NO: 6, an HCDR2 of SEQ ID NO: 7, and an HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3.In another embodiment, the hybridoma producing the anti-ASC monoclonal antibody of the present disclosure produces a monoclonal antibody comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3. In yet another embodiment, the hybridoma producing the anti-ASC monoclonal antibody of the present disclosure produces a monoclonal antibody comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the amino acid sequence of the VH region comprises HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3, and the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3.

[0129] Administration of the Composition The compositions of the present invention can be administered to mammals (e.g., rodents, humans) in any suitable formulation. For example, anti-ASC antibodies can be formulated in a pharmaceutically acceptable carrier or diluent, such as physiological saline or buffered saline solution. Suitable carriers and diluents can be selected based on the mode and route of administration and standard pharmaceutical practice. Exemplary pharmaceutically acceptable carriers and diluents, as well as descriptions of pharmaceutical formulations, can be found in Remington's Pharmaceutical Sciences and USP / NF, standard texts in this field. Other substances may be added to the composition to stabilize and / or preserve the composition.

[0130] The compositions of the present invention can be administered to mammals by any conventional technique. Typically, such administration will be by inhalation, intranasal, or parenteral (e.g., intravenous, subcutaneous, intratumoral, intramuscular, intraperitoneal, or intrathecal introduction). The compositions may also be administered directly to the target site, for example, by surgical delivery to an internal or external target site or by a catheter to a site accessible by blood vessels. The compositions may be administered by a single bolus, multiple injections, or continuous infusion (e.g., intravenous, peritoneal dialysis, pump infusion). For inhalation or intranasal administration, the compositions may be administered using an inhaler, nebulizer, or any suitable device. For parenteral administration, the compositions may be formulated in a sterile, pyrogen-free form.

[0131] Effective dose The compositions described above can be administered to a mammal (e.g., a rat, a human) in an effective amount, i.e., an amount capable of producing the desired result in the treated mammal (e.g., reducing inflammation in the CNS of a mammal that has suffered a traumatic injury to the CNS or a stroke, or a mammal with an autoimmune disease or a CNS disorder). Such a therapeutically effective amount can be determined as described below. A therapeutically effective amount of a composition comprising an agent provided herein (e.g., a monoclonal antibody provided herein or an antibody fragment derived therefrom, e.g., IC-100) can generally be about 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 mg / kg of patient body weight. A therapeutically effective amount of a composition comprising an agent provided herein (e.g., a monoclonal antibody provided herein or an antibody fragment derived therefrom, e.g., IC-100) can generally be from about 0.001 to about 200 mg / kg of patient body weight. A therapeutically effective amount of a composition comprising an agent provided herein (e.g., a monoclonal antibody provided herein or an antibody fragment derived therefrom, e.g., IC-100) can generally be about 0.001 mg / kg to about 0.01 mg / kg, about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 1 mg / kg to about 10 mg / kg, about 10 mg / kg to about 25 mg / kg, about 25 mg / kg to about 50 mg / kg, about 50 mg / kg to about 75 mg / kg, about 75 mg / kg to about 100 mg / kg, about 100 mg / kg to about 125 mg / kg, about 125 mg / kg to about 150 mg / kg, about 150 mg / kg to about 175 mg / kg, or about 175 mg / kg to about 200 mg / kg of patient body weight. Compositions containing agents provided herein (monoclonal antibodies provided herein or antibody fragments derived therefrom, e.g., IC-100) can be administered in single or multiple doses.

[0132] The toxicity and therapeutic efficacy of the compositions used in the methods of the present invention are measured using LD 50 The LD (the dose at which 50% of a population is lethal) can be determined by standard pharmaceutical procedures using either cultured cells or experimental animals. The dose ratio between toxic and therapeutic effects is the therapeutic index, which is known as the LD 50 / ED 50 In some cases, the compositions provided herein exhibit a large therapeutic index. While those that exhibit toxic side effects may be used, care should be taken to design a delivery system that minimizes the potential damage of such side effects. In some cases, the dosage of the compositions provided herein may be such that the ED 50 The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.

[0133] As is well known in human and veterinary medicine, the dosage for any one subject will depend on many factors, including the subject's size, body surface area, age, the particular composition being administered, the time and route of administration, general health, and other medications being administered concomitantly. [Example]

[0134] Example The present invention is further described by the following specific examples, which are provided for illustrative purposes only and should not be construed as limiting the scope of the invention in any way.

[0135] Example 1: The role of EV-mediated inflammasome signaling in ALI after TBI and the effect of its neutralization Pulmonary dysfunction is a common complication of severe traumatic brain injury. Approximately 20–25% of TBI subjects develop acute lung injury (ALI), but the mechanisms mediating the pathogenesis of TBI-induced ALI remain unclear. Previous literature has supported the view that post-TBI pulmonary dysfunction is due to cardiopulmonary dysfunction resulting from the sympathetic nervous response to increased intracranial pressure. However, more recent studies have demonstrated that the systemic inflammatory response also plays an important role in TBI-induced lung injury. Specifically, the HMGB1-RAGE ligand receptor pathway serves as a central mediator of post-TBI pulmonary dysfunction. Furthermore, HMGB1 induces activation of the AIM2 inflammasome. Furthermore, previous literature has revealed that pathogens secrete EVs carrying DAMPs, such as HMGB1, to trigger inflammation (Buzas et al., 2014). Various studies have shown that after TBI, the blood-brain barrier (BBB) ​​becomes permeable as early as 3–6 hours after injury, damaging the protective barrier between the brain and intravascular compartments and resulting in the leakage of proteins and fluids. When the BBB is disrupted after injury, inflammatory mediators such as DAMPs are secreted, which can promote brain inflammation and damage distant organs. Although some inflammatory mediators function as clear markers of brain injury, their validity is not generally recognized. Furthermore, there are currently no clinically approved treatments or biomarkers for TBI-induced ALI. In recent years, EVs have become a hot area of ​​research in biomarkers for several different types of diseases, including lung injury and TBI. EVs isolated from the cerebrospinal fluid of TBI patients have previously been shown to have increased inflammasome proteins compared to control samples. In this example, we examined the contribution of EV-mediated inflammasome signaling in the pathogenesis of TBI-induced ALI.

[0136] Materials and Methods Animals and traumatic brain injury All animal experiments were approved by the Institutional Animal Care and Use Committee of the University of Miami Miller School of Medicine (Animal Welfare Assurance A3224-01) and conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals. The ARRIVE guidelines were followed in conducting this study. All C57 / BL6 mice were 8-12 weeks old and weighed 24-32 grams. Mice were pre-randomized into TBI experimental groups (sham, 4 h, 24 h) and adoptive transfer and treatment experimental groups (naive, sham-saline, untreated, enoxaparin, anti-ASC). For the TBI experimental group, sham animals underwent surgery but were not injured. For the adoptive transfer treatment study, the sham-saline group underwent surgery and received saline as vehicle treatment. Naive animals did not undergo surgery. Based on power analysis (effect size F = 0.85, with α set at 0.05 using G* power analysis) and previous data, a sample size of 5–6 mice per group was used. All mice were housed in a viral antigen-free (VAF) animal facility at the University of Miami Lois Pope Life Center under a 12-hour light-dark cycle with free access to food and water. The facility performed husbandry procedures twice weekly and checked the animals daily. After surgery, animals were observed in the operating room while being maintained on a heating pad and their body temperature controlled with a rectal probe, maintained at 37°C, and then transferred to the animal room.

[0137] Prior to surgery, animals were anesthetized with ketamine and xylazine (intraperitoneally, i.p.). The anesthetized animals were then placed on a heating pad to maintain a body temperature of 37°C. TBI was performed using the Controlled Cortical Impact (CCI) model. A 5 mm craniotomy was performed in the right cortex (-2.5 mm posterior and 2.0 mm lateral from bregma). Injury was induced using an ECCI-6.3 device (Custom Design & Fabrication, Richmond, Va., USA) with a 3 mm impounder at a velocity of 6 m / s, a depth of 0.8 mm, and an impact duration of 150 m. After these procedures, animals were returned to their cages and provided with food and water. Animals were sacrificed 4 and 24 hours after TBI, as described. Sham animals were anesthetized and underwent the same preoperative incisions as the injured animals, but did not undergo a craniotomy or contusion.

[0138] Tissue collection All animals were anesthetized with ketamine and xylazine before perfusion. Then, animals underwent tracheal perfusion. Using a 20 cmH2O tracheal catheter, 4% paraformaldehyde (PFA) was instilled into the lungs, followed by fixation in 4% PFA overnight at 4°C. Fixed lung tissue was embedded in paraffin and processed into 5-m sections. Right lung tissue was harvested for protein isolation and molecular analysis. Subsequently, animals were decapitated, and right cortical tissue was harvested for protein isolation and molecular analysis.

[0139] Pyrosome isolation assay Mouse lung tissue lysates were filtered through a 5 μm low-binding polyvinylidene fluoride (PVDF) membrane (Millipore). After filtration, the supernatant was centrifuged at 2,700 × g for 8 minutes. The pellet was resuspended in 40 μl of 3[(3-cholamidopropyl)dimethylammonio]-propanesulfonic acid (CHAPS) buffer (20 mmol / L HEPES-KOH, pH 7.5, 5 mmol / L MgCl2, 0.5 mmol / L EGTA, 0.1 mmol / L phenylmethylsulfonyl fluoride, protease inhibitor cocktail, 0.1% CHAPS). Pyroptosomes were pelleted by centrifugation at 2,700 × g for 8 minutes. The pellet was resuspended and incubated in 27.8 μl of CHAPS buffer containing 2.2 μl of disuccinimidyl substrate at room temperature for 30 minutes to crosslink ASC dimers. Finally, an equal volume of 2× Laemmli buffer was added, and proteins were analyzed by immunoblotting using commercially available antibodies against ASC and gasdermin D (GSD).

[0140] Nuclear and cytoplasmic extraction Nuclear and cytoplasmic fractions were extracted using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific) according to the manufacturer's instructions. Briefly, mouse lung tissue samples were cut into 20–100 mg sections and centrifuged at 500 × g for 5 minutes. The tissue pieces were then homogenized in cytoplasmic extraction reagent and centrifuged at 16,000 × g for 5 minutes. The supernatant (cell extract) was then removed, and the pellet was centrifuged with Nuclear Extraction Reagent (Thermo Scientific) at 16,000 × g for 10 minutes. This supernatant, which corresponds to the nuclear fraction, was removed and stored at -80°C.

[0141] Immunoblotting Lung and brain tissue samples were snap-frozen in liquid nitrogen and stored at -80°C. Two-mm sections of right lower lung and right cortical tissue were homogenized in extraction buffer containing a protease and phosphatase inhibitor cocktail (Sigma, St. Louis, MO, USA) and separated in 4-20% Tris-TGX Criterion precast gels (Bio-Rad, Hercules, CA, USA) using antibodies against caspase-1 (Novus Biologicals), ASC (Santa Cruz), IL-1 (Cell Signaling), IL-18 (Abcam), AIM2 (Santa Cruz), and HMGB1 (Millipore) as described by de Rivero Vaccari et al. (2015). Band density was quantified using Image Lab, and all data were normalized to 0-actin.

[0142] immunohistochemistry Tissue sections were deparaffinized with xylene and then rehydrated with ethanol and Tris-buffered saline. Immunohistochemical procedures for double staining were then performed as previously described. Sections were incubated overnight at 4°C with antibodies against caspase-1 and ASC (Millipore), AIM2 (Santa Cruz), HMGB1 (Millipore), and SPC (Millipore). Immunostained lung sections from sham, 4-hour, and 24-hour mice were examined with a Zeiss laser scanning confocal microscope (Zeiss, Inc., Thornwood, NY, USA). Lung sections were analyzed by individuals blinded to each group.

[0143] EV isolation EVs were isolated from serum of TBI-injured and injured mice using Total Exosome Isolation Solution according to the manufacturer's instructions (Invitrogen). Briefly, 100 μl of each sample was centrifuged at 2000 × g for 30 minutes. The supernatant was incubated with 20 μl of Total Exosome Isolation (TEI) reagent at 4°C for 30 minutes and then centrifuged at 10,000 × g for 10 minutes at room temperature. The supernatant was discarded, and the pellet was resuspended in 100 μl of PBS. EVs were characterized by CD81 expression and Nanosight tracking analysis (Figure 6).

[0144] Adoptive transfer of EVs Serum-derived EVs from C57BL-6TBI mice and sham mice were collected at 1.0 x 10 per gram / body weight. 10 A dose of 1000 particles was injected into naive C57BL-6 mice via the jugular vein. Particle counts were measured using Nanosight Tracking analysis, and samples were diluted appropriately. Prior to surgery, animals were anesthetized with ketamine and xylene. A 1-2 cm incision was made between the chin and clavicle. The jugular vein was then pulled up and tied off, after which a catheter was placed. Serum-derived EVs were transplanted, and lung and brain tissues were collected and analyzed 24 hours after injection (n=5).

[0145] Enoxaparin and anti-ASC treatment Serum-derived EVs from TBI mice were injected into naive C57-BL6 mice via the jugular vein. One hour later, enoxaparin (3 mg / kg) (n = 4) and anti-ASC (5 mg / kg) (n = 4) were administered to recipient animals. The following groups were used: 1) the naive group, which received no treatment; 2) the sham saline group, which served as a negative control and received saline alone via the jugular vein; 3) the untreated group, which received EVs from TBI mice without treatment and served as a positive control; 4) the ENOX group, which received EVs from TBI mice and enoxaparin; and 5) the anti-ASC group, which received EVs from TBI mice and anti-ASC. The treatment order was randomized. 24 hours after injection, lung and brain tissues were collected and analyzed. The anti-ASC antibody used in the treatment experiments was a humanized monoclonal antibody against ASC, which recognizes mouse, human, and porcine ASCs.

[0146] Histology and lung injury scoring Lung tissue sections were stained with standard hematoxylin and eosin for histology, morphology, and ALI scoring. Lung sections were scored by a blinded pathologist using the lung injury scoring system from the American Thoracic Society Workshop Report. Twenty random high-power fields were selected for scoring. ALI scoring criteria were based on the number of neutrophils in the alveolar space, the presence of interstitial spaces, hyaline membranes, protein debris filling the airspaces, and thickening of the alveolar septa. Based on these criteria, a score ranging from 0 (no injury) to 1 (severe injury) was assigned.

[0147] statistical analysis Data were analyzed using Student's t-test for two groups and one-way ANOVA for more than two groups, followed by Tukey's multiple comparison test (GraphPad Prism version 7.0). Normality was verified using the D'Agostino-Pearson test. Data are expressed as mean ± SEM. The P value used for significance was *p<0.05.

[0148] result Severe TBI increases AIM2 inflammasome protein and HMGB1 expression in the mouse brain Excessive levels of inflammatory cytokines IL-1β and IL-18, as well as inflammasome proteins, are associated with secondary injury after fluid percussion brain injury. Cortical lysates were analyzed to determine whether severe CCI induces changes in inflammatory cytokine processing and inflammasome protein levels, but research on inflammasome activation in severe TBI is limited. In this example, we examined the levels of caspase-1 (Figure 1A, B) (p<0.001), ASC (Figure 1A, C) (p=0.003), IL-18 (Figure 1A, D) (p=0.0042), AIM2 (Figure 1A, F) (p=0.0197), and IL-1β (Figure 1A, G) (p=0.0141) in cortical lysates 4 and 24 hours after injury after severe CCI. Levels of caspase-1, ASC, AIM2, and IL-1β peaked 4 hours after CCI and declined by 24 hours. The time course of inflammatory cytokine maturation varied slightly, but peaked by 24 hours after TBI. Because other studies have demonstrated the role of the inflammasome DAMP HMGB1 in activating the AIM2 inflammasome, we also measured the levels of these proteins in cortical lysates. As shown in Figures 1A and 1E, CCI induced a significant increase in HMGB1 (Figures 1A and 1E) levels at 4 and 24 hours after injury (p = 0.0121). These data indicate that after severe CCI in mice, the levels of the AIM2 inflammasome protein were significantly elevated in the injured cerebral cortex.

[0149] Severe TBI increases the expression of AIM2 inflammasome protein and HMGB1 in mouse lungs To determine whether CCI induced inflammasome activation in the lung, we performed immunoblot analysis of lung lysates for caspase-1 (Figure 1H,I) (p = 0.0026), ASC (Figure 1H,J) (p = 0.0427), IL-18 (Figure 1H,K) (p = 0.0025), IL-1β (Figure 1H,N) (p = 0.0012), and AIM2 (Figure 1H,M) (p < 0.001), as well as NLRP3 (p = 0.0047) (Supplementary Figure 1). Increased levels of caspase-1, ASC, IL-18, and AIM2 were significantly increased at 4 and 24 h after injury compared with sham controls. However, the time course of increased protein expression was somewhat different from that observed in the brain, which peaked at 24 h after CCI. Because the HMGB1-RAGE axis is involved in the mechanism by which TBI causes lung dysfunction, lung lysates were analyzed for HMGB1 protein expression levels. Figures 1H and 1L (p=0.0158) show that HMGB1 expression increased 4 and 24 hours after TBI, indicating that the AIM2 inflammasome and HMGB1 are involved in the lung inflammatory response after TBI.

[0150] TBI induces pyroptosis in the lungs of mice As previously shown, activation of the AIM2 inflammasome in cortical neurons triggers pyroptotic cell death. To investigate whether TBI induces pyroptosis in mouse lung tissue, pyrosomes were isolated from lung tissue after TBI. TBI animals sacrificed 4 h after injury showed evidence of ASC oligomerization compared with sham animals (Figure 4A). ASC dimers and trimers (50 and 75 kDA, respectively) were observed in TBI animals. These results indicated pyrosome formation, characterized by supramolecular assemblies of ASC oligomers. Furthermore, gasdermin D (GSDMD), which is cleaved upon caspase-1 activation and triggers pyroptosis and IL-1β release, was significantly increased in the lungs of TBI animals compared with sham animals (Figures 4B and 4C) (p = 0.0001). These observations indicate that pyroptosis contributes to lung cell death after TBI.

[0151] TBI increases inflammasome protein immunoreactivity in type II alveolar epithelial cells TBI leads to capillary leakage, resulting in increased vascular permeability and potentially damaging specialized alveolar epithelial cells known as type II alveoli. To examine the cellular effects of TBI on inflammasome expression in the lung after injury, we performed immunohistochemical analysis on lung sections from sham, 4-hour, and 24-hour injured animals. Type II alveolar epithelial cells are known to be the predominant type of lung cell damaged in ALI. Lung sections were stained with antibodies against AIM2, caspase-1, and ASC (green) and co-stained with pro-surfactant protein C (Pro-SPC, red), a marker for type II epithelial cells, and DAPI nuclear stain (blue). As shown in Figures 2A–2C, active caspase-1 (Figure 2A), ASC (Figure 2B), and AIM2 (Figure 2C) are present in SPC-positive cells (arrows). The immunoreactivity of these inflammasome proteins increased after TBI. These results indicate that type II alveolar epithelial cells express inflammasome proteins and that TBI induces increased immune reactivity in these cells.

[0152] TBI increases nuclear and cytoplasmic HMGB1 expression To determine the cellular distribution of HMGB1 in lung cells after TBI, nuclear and cytoplasmic fractions were isolated from lung homogenates (Figures 3A and 3C) (p = 0.0337). Immunoblotting showed a significant increase in HMGB1 expression in both fractions at 4 h after TBI (Figures 3B and 3D) (p = 0.0345). To examine changes in immunoreactivity in lung sections after TBI, immunohistochemical analysis of HMGB1 was also performed. Sections were co-stained with HMGB1 (green) and SPC (red) and DAPI nuclear stain (blue). HMGB1 immunoreactivity increased at 4 and 24 h compared with sham controls. Weak HMGB1 immunoreactivity was observed in SPC-positive cells (arrows) (Figure 3E); therefore, it is suggested that changes in HMGB1 in injured lung tissue may be cytoplasmic.

[0153] TBI induces morphological changes in the lungs and induces ALI ALI is characterized by an inflammatory process that causes alveolar and interstitial edema and infiltration of inflammatory cells into the alveolar space. Histopathological analysis of lung tissue (Figure 5A) demonstrates that severe TBI induces substantial changes in lung structure and morphology at 4 and 24 hours after injury. Sham animals exhibited normal alveolar morphology, whereas injured animals showed acute changes in alveolar edema, which slightly decreased by 24 hours after injury (long arrows). Furthermore, evidence of neutrophil infiltration (arrowheads) and morphological changes in the alveolar-capillary membrane (*) was observed at both time points. Injured animals showed signs of interstitial edema, which was more pronounced at 4 hours after injury but was still evident at 24 hours after injury (short arrows). Finally, injured animals also showed evidence of thickening of the interstitial areas and alveolar septa (lb, #).

[0154] To confirm that severe injury induces ALI, tissue sections were analyzed using the ALI scoring system established by the American Thoracic Society. This system is based on evidence of neutrophil infiltration into the alveolar and interstitial spaces, hyaline membrane formation, protein debris filling the airspaces, and thickening of the alveolar septa. These features were significantly elevated in injured animals, and ALI scores were overall higher in TBI animals compared to sham animals (Figure 5B) (p = 0.0017).

[0155] Enoxaparin and anti-ASC antibody treatment significantly reduced inflammasome expression and ALI after adoptive transfer of EVs from TBI mice To obtain evidence that EVs and their cargoes, which may be released into the circulation after TBI, can induce inflammasome activation in the lung, we performed a classical adoptive transfer experiment using serum-derived EVs from mice with severe CCI. EV preparations were validated using Western blots for the EV marker CD81 (Figure 6). Controls received EVs isolated from sham or naive animals. As shown in Figures 7A–7F, active caspase-1 (Figures 7A, 7B), ASC (Figures 7A, 7C), IL-18 (Figures 7A, 7D), AIM2 (Figures 7A, 7E), and HMGB1 (Figures 7A, 7F) were significantly elevated in the lungs of animals receiving EVs from TBI-injured animals compared with the lungs of animals receiving EVs from non-injured or naive mice. Furthermore, in the lungs treated with EVs from TBI mice, infiltration of inflammatory cells (arrows) was evident (Figure 7G). Finally, ALI scores were also significantly higher in animals receiving EVs from injured mice (Figure 7G). These studies provided evidence of a neuro-respiratory-inflammasome axis in which EVs released into the circulation after TBI activate inflammasomes in target cells in the lung, which contributes to the pathogenesis of ALI.

[0156] Next, we attempted to block exosome uptake by treating naive mice with either enoxaparin or a monoclonal antibody against ASC after adoptive transfer of EVs from injured mice. Negative control animals received saline, and positive control animals received no treatment. As shown in Figures 8A–8F, caspase-1 (Figures 8A, 8B), ASC (Figures 8A, 8C), TL-1β (Figures 8A, 8D), AIM2 (Figures 8A, 8E), and HMGB1 (Figures 8A, 8F) were significantly reduced after treatment with enoxaparin or a humanized anti-ASC monoclonal antibody (e.g., IC100 antibody) compared with the untreated (positive control) group (p = <0.0001). Furthermore, H&E-stained lung sections showed significantly less neutrophil infiltration into the alveolar and interstitial spaces, with no signs of septal thickening (Figures 9A–D). The ALI scores of animals treated with enoxaparin and anti-ASC antibody (IC100) were significantly lower than those of the untreated group (Figure 9E) (p=<0.0001). Thus, EVs released into the circulation after TBI are involved in inflammasome activation in lung cells, leading to ALI.

[0157] conclusion TBI may be associated with a higher rate of certain medical complications, particularly pulmonary and central nervous system dysfunction. This study demonstrates that severe TBI increases HMGB1 and inflammasome expression (e.g., AIM2, caspase-1, and ASC expression) in cortical and lung tissue and induces lung morphological changes consistent with ALI (e.g., neutrophil infiltration into alveolar and interstitial spaces, alveolar septal thickening, and alveolar edema and hemorrhage), supporting the concept of a neurorespiratory inflammation axis. Importantly, TBI induces pyroptosis in lung tissue (e.g., the presence of GSDMD cleavage) and increases the expression of inflammasome proteins in type II alveolar epithelial cells. Furthermore, adoptive transfer of EVs from TBI mice activated inflammasomes and induced ALI, indicating that brain injury induces the release of EVs containing inflammasome protein cargo, which in turn leads to ALI. Furthermore, it was shown that both inhibition of EV uptake (enoxaparin) and inflammasome activation (treatment with anti-ASC antibody (IC100)) resulted in suppression of inflammasome protein expression and the development of ALI.

[0158] In summary, this example demonstrated that AIM2 inflammasome signaling plays a central role in the pathogenesis of lung injury after TBI and demonstrated a mechanism of TBI-induced ALI involving EV-mediated inflammasome signaling. These data provided evidence that EV-mediated inflammasome signaling may play a central role in mediating the neuro-respiratory-inflammatory axis. Therefore, targeting this axis with antibodies against inflammasome proteins or drugs that block EV uptake could provide a therapeutic approach for neurotraumatic ALI in all fields of intensive care medicine. These results suggest that the disclosed therapeutic strategy may be useful for the treatment of pulmonary inflammatory diseases in general.

[0159] Example 2: Role of EV-mediated inflammasome signaling in ALI after TBI in human patients As a follow-up to the mouse experiments in Example 1, we investigated the role of EVs isolated from human TBI patients on inflammasome signaling in human pulmonary endothelial cells.

[0160] In the first experiment, serum-derived EVs were isolated from TBI and control patients using the Total Exosome Isolation Kit (ThermoFisher). Human pulmonary microvascular endothelial cells (HMVEC-Lonza) were cultured and plated in 12-well plates. After reaching confluence, EVs isolated from TBI and control patients were delivered to the cells (1.94 × 10 particles / ml) and incubated for 4 hours. After incubation, cells were harvested using 200 μl of lysis buffer, and cell lysates were used for Western blot analysis.

[0161] In the second experiment, serum-derived EVs were isolated from TBI and control patients using the Total Exosome Isolation Kit (ThermoFisher). Human pulmonary microvascular endothelial cells (HMVEC-Lonza) were cultured and plated in 96-well plates. After reaching confluence, EVs isolated from TBI and control patients were delivered to the cells (1.94 × 10 particles / ml). After 3 hours of incubation, caspase-1 FAM FLICA (Immunohistochemistry Technologies) was added at a volume ratio of 1:30 for an additional hour. After incubation, the medium was removed, and the cells were washed three times with apoptosis wash buffer (Immunohistochemistry Technologies). Cells were then co-stained with Hoechst for nuclear staining and propidium iodide for cell death. Images were captured using an EVOS microscope, and cells were read using a fluorescence plate reader at an excitation wavelength of 492 nm and an emission wavelength of 520 nm.

[0162] result As shown in Figures 10A-10F, delivery of serum-derived EVs from TBI patients increased the expression of inflammasome proteins in lung endothelial cells. Figures 10A-10E showed that PMVECs incubated with TBI-EVs for 4 hours showed elevated levels of caspase-1, ASC, AIM2, and HMGB1 compared with PMVECs incubated with control-EVs for 4 hours. Immunoassay results showed a significant increase in IL-1β expression using the Ella simple plex assay (Figure 10F).

[0163] As shown in Figures 11A-11C, delivery of TBI-EVs to pulmonary endothelial cells increased caspase-1 immunoreactivity and cell death.

[0164] conclusion These studies provided further evidence for the neuro-respiratory-inflammasome axis, in which EVs released into the circulation after TBI activate inflammasomes in target cells in the lung and contribute to the pathogenesis of ALI.

[0165] Example 3: Effect of the use of humanized anti-ASC antibodies in an animal model of multiple sclerosis To determine the utility of a humanized anti-ASC monoclonal antibody in treating MS, the antibody was administered to mice with experimental allergic encephalomyelitis (EAE). EAE is an animal (i.e., rodent) model of MS, as described in Hoeftberger R, Leisser M, Bauer J, Lassmann H (December 2015). "Autoimmune encephalitis in humans: how closely does it reflect multiple sclerosis?" Acta Neuropathol. Commun. 3(1): 80; and Lassmann Hans (February 2010). "Acute disseminated encephalomyelitis and multiple sclerosis." Brain. 133: 317-319; and L. Gomez Vicente et al. Relapse in a paucisymptomatic form of multiple sclerosis in a patient treated with nivolumab, Neuro Oncol (2016) 18 (suppl 4): iv25.

[0166] method Induction of EAE and treatment with IC-100 Active EAE was induced in 2-month-old C57BL / 6 female mice by injecting myelin oligodendrocyte glycoprotein 35-55 peptide (MOG 35-55 , BioSynthesis) as previously described (Brambilla et al., 2014). Briefly, mice received an intraperitoneal (ip) injection of pertussis toxin (dissolved in PBS (350 ng / mouse; day 0)) followed by MOG emulsified in complete Freund's adjuvant. 35-55Mice received a subcutaneous injection of pertussis toxin (300 ng / mouse; day 1), followed by a second ip injection of pertussis toxin (350 ng / mouse; day 2). Mice were injected ip with vehicle (0.9% saline) or IC-100 at three different doses (10, 30, and 45 mg / kg) every four days starting on day 8 after EAE induction. Clinical signs of EAE were scored daily on a scale of 0 to 6 as follows: 0, no clinical signs; 1, loss of tail tone; 2, flaccid tail; 3, complete hindlimb paralysis; 4, complete forelimb paralysis; 5, moribund; and 6, death.

[0167] Cell isolation for flow cytometry After transcardial perfusion with PBS, the spinal cord was harvested and Mg 2+ and Ca 2+ The cells were then introduced into cold Hanks' Balanced Salt Solution (HBSS w / o) without ATP. The samples were manually dissociated into a single-cell suspension using a 70-um strainer and washed with HBSS w / o. The spleen samples were spun at 1200 rpm for 10 minutes at 4°C, the supernatant removed, and red blood cells (RBCs) were lysed in 2 ml of RBC lysis buffer (eBioscience) according to the manufacturer's instructions. The spleen cells were then resuspended in PBS. Cells isolated from the spinal cord were suspended in flow cytometry buffer (FCB, eBioscience) and incubated with Myelin Removal Beads II (Miltenyi). Myelin was removed using an LS magnetic column as described in the manufacturer's protocol (Miltenyi). Similar to the splenocytes, the spinal cord cells were also resuspended in PBS and stained as described below.

[0168] Immunolabeling and flow cytometry analysis The FAM FLICA™ Caspase 1 Kit (BioRad) was used. Cells were incubated in FLICA solution (BioRad) for 30 minutes at 4°C, washed with Apoptosis Wash Buffer (BioRad), and resuspended in 1 ml of PBS. Samples were then incubated with fixable viability reagent (Tonbo Biosciences) for 30 minutes at 4°C, spun at 1200 rpm for 10 minutes at 4°C, and the supernatant was removed. Cells were resuspended in 100 μl of FACS buffer, blocked with anti-CD16 / 32 (FcR block, eBioscience) for 5 minutes at room temperature, immunostained for 30 minutes at 4°C, and then fixed with 1% PFA. Samples were analyzed using a CytoFLEX S flow cytometer equipped with CytExpert 2.1 software (Beckman Coulter). Spinal cord leukocyte counts were measured using 123count eBeads (eBioscience). Splenic leukocyte counts were determined by flow cytometry combined with trypan blue exclusion counting using a TC20™ Automated Cell Counter (Bio-Rad). A list of flow cytometry antibodies is provided in Table 3 below.

[0169] [Table 8]

[0170] Luxol fast blue staining and quantification of demyelinated white matter volume Paraformaldehyde (PFA)-fixed sections of the spinal cord were paraffin-embedded, cut into 10-mm-thick cross sections using a Leica RM 2135 microtome, and stained with Luxol Fast Blue (LFB). Ten serial sections spaced 50 μm apart were used to estimate the volume of demyelinated white matter. The demyelinated areas were outlined using an Olympus BX51 microscope, and the volume of demyelinated white matter was quantified using Stereoinvestigator software (MicroBrightfield). 3D reconstruction of the demyelinated spinal cord was performed on the same serial sections using Neurolucida software (MBF Bioscience).

[0171] result Treatment with anti-ASC antibody IC-100 improves functional outcome in experimental autoimmune encephalomyelitis (EAE) To evaluate the therapeutic potential of IC-100, 2-month-old female C57BL / 6 mice were treated with MOG 35-55 EAE was induced using a peptide (Brambilla et al., 2014), and IC-100 or vehicle alone was administered starting 8 days post-induction of the disease (dpi). Administration was repeated every 4 days until sacrifice (set at 35 dpi). Three doses were administered: 10, 30, and 45 mg / kg.

[0172] IC-100, when used at doses of 30 mg / Kg and 45 mg / Kg, significantly improved functional recovery and resulted in a robust reduction in clinical disease scores over the course of the experiment (Figure 12A). Treatment reduced the mean peak clinical score (Figure 12B) and the overall severity of EAE, measured as a reduction in the cumulative disease index (CDI) (Figure 12C). Mice treated with 30 mg / Kg and 45 mg / Kg of IC-100 also showed a trend toward delayed disease onset (Figure 12D). No differences were observed in the day on which mice reached peak disease scores (Figure 12E).

[0173] Treatment with the anti-ASC antibody IC-100 suppresses peripheral immune cell infiltration into the spinal cord after EAE The onset, duration, and severity of EAE clinical symptoms directly correlate with immune cell infiltration into the spinal cord. To assess whether IC-100 influences this process, immune cell populations isolated from spinal cords at 35 dpi were profiled by flow cytometry. Administration of 30 mg / kg of IC-100 significantly reduced brain-derived CD4+ T cells, the most important immune cell populations in driving EAE pathology, as well as total CD8+ T cells (Figure 13A). All other immune cell populations showed a clear trend toward reduction. No differences in cell numbers were observed in the spleen at any dose of IC-100, suggesting that the treatment did not interfere with the ability of mice to mount a sufficient immune response to EAE challenge (Figure 13B).

[0174] Treatment with anti-ASC antibody IC-100 reduces the number and activation state of microglia after EAE Microglia are involved in the immune-inflammatory response to CNS diseases. Increased activation of microglia leads to proliferation and increased surface expression of MHCII. To assess whether IC-100 influences this response, we measured the total number of microglia and MHCII expression in the spinal cord. + The number of activated microglia was quantified by flow cytometry. Both populations were significantly reduced by treatment with 30 mg / kg of IC-100, indicating that at this dose, IC-100 is effective in suppressing microglial activation and microglial-mediated neuroinflammation (see Figure 14).

[0175] Example 4: Use of humanized anti-ASC antibodies in an animal model of virus-associated pneumonia We use a murine coronavirus, mouse hepatitis virus strain A59 (MHV-A59), as a model to study the pathogenesis of coronavirus infection and the host response to infection. This model virus has been shown to infect the lungs, liver, spleen, brain, and spinal cord, among other organs. In addition, this model also exhibits demyelination consistent with a multiple sclerosis-like phenotype (Weiss and Leibowitz 2011). Therefore, we propose to use the MHV-A59 model to test the effect of IC100 in ameliorating the effects of coronavirus infection on survival / recovery and inflammatory cytokine production. We hypothesize that IC100 binds to ASC and inhibits inflammasome activation as a result of viral infection, thereby increasing survival and reducing the production of inflammatory cytokines, including IL-1β, TNF, and IL-6.

[0176] Example 5: Diagnosis and Treatment of Alzheimer's Disease Materials and Methods Postmortem human brain Informed consent was obtained for the postmortem examination study in accordance with the guidelines of the University of Miami Institutional Regulatory Board (IRB). Research ethics were obtained from the Human Subjects Research Office, University of Miami, Miami, Florida (IRB ethics number, 19920348 (CR00012340) Brain Endowment Bank).

[0177] tissue preparation This study used 17 postmortem brains from donors with intermediate AD (intermediate AD neuropathological changes, mean age 83.41 ± 7.05 years; 10 women and 7 men) and 17 age-matched non-demented controls with age-related neuropathological changes (low AD, mean age 84.59 ± 10.26 years; 10 women and 7 men). Diagnosis of cognitive status was made by a clinician noted in the patient's medical record and reassessed by a cognitive neurologist (XS). Cases with clinical findings of neurodegenerative diseases other than AD were excluded from the study. The primary cause of death for each case was indicated by the clinician, and the level of AD neuropathological changes was confirmed by a neuropathologist (SHG). Briefly, whole brains were harvested an average of 17.23 ± 8.72 hours after death. The left hemisphere was frozen, and the right hemisphere was fixed in 10% formalin (pH 7.0) for 1 month, then sliced, sampled, and embedded in paraffin blocks processed with a Leica Tissue-Tek Processor (Leica Biosystems, Buffalo Grove, IL, USA). Paraffin-embedded tissue sections were then obtained from the hippocampal formation, including the entorhinal cortex, cornu ammonis (CA1-CA3), subiculum, and dentate gyrus (DG), for immunohistochemical and histochemical staining, as described below. The paraffin-embedded tissue blocks were cut at 20 μm thickness using a Leica RM2245 microtome (Leica Microsystems Ltd.), and 20 serial sections were collected. For further validation, three sections were selected from each block based on the principle of systematic extraction and a 1:5 section extraction ratio.

[0178] Immunohistochemistry and histochemistry Standard immunohistochemical procedures for 20-μm-thick brain sections have been described previously. Briefly, after deparaffinization, endogenous peroxidase activity was quenched by placing slides in 3% hydrogen peroxide (HO) for 10 min. Sections were immersed in preheated 10 mM citric acid, pH 6.0 (VWR., Radnor, PA, USA) for 30 min, cooled in ice water, or pretreated with formic acid (Sigma-Aldrich, St. Louis, MO, USA) for 5 min. Next, sections were blocked with 5% goat serum (Vector Laboratories, Burlingame, Calif., USA) for 20 min and then incubated with mouse anti-phosphotau (Ser202, Thr205) antibody (AT-8; 0.4 μg / mL; ThermoFisher Chemicals, Richmond, VA, USA), mouse anti-β-amyloid (6E10; 1.0 μg / mL; Biolegend, San Diego, CA, USA), rabbit anti-NLRP3 (3.0 μg / mL; MilliporeSigma, St. Louis, MO, USA), mouse anti-NLRP1 (1.0 μg / mL; Enzo Life Sciences, Farmingdale, NY, USA), and mouse anti-ASC (B-3; Santa Cruz Biotechnology, Santa Cruz, CA, USA), as described. Sections were incubated overnight at 4°C in PBS with a solution of primary antibodies (0.4 μg / mL), human anti-ASC (IC100, 2 μg / mL), and rabbit anti-caspase 1 (0.6 μg / mL; MilliporeSigma). The next day, sections were exposed to biotinylated horse anti-mouse IgG or biotinylated goat anti-rabbit IgG secondary antibodies (15 μg / mL; Vector Laboratories) in PBS for 1 h, followed by avidin-biotin complex (1:200, ABC; Vector Laboratories) for 1 h. The reaction was visualized with 3,3'-diaminobenzidine (MilliporeSigma) for 10 min. Finally, sections were dehydrated, cleared with xylene, and coverslipped. Staining was performed in the absence of primary antibodies (as a negative low-concentration AD), but no specific staining was observed with these preparations.Furthermore, we compared immunoreactivity using skin and tonsil samples as positive low-AD samples. https: / / www.proteinatlas.org / (Supplementary Data, Figure S1).

[0179] Microscopic analysis Hematoxylin and eosin (H&E) staining was used to assess the general morphology of brain tissue and the orientation of brain regions, as previously described. For H&E staining, standard histological paraffin blocks were cut at 20 μm thickness, and the slides were dried and then heated at 60 °C for 30 min. Prior to staining, sections were deparaffinized in three changes of xylene and rehydrated through graded concentrations of ethanol. A strategy for identifying sampling sites and regions of interest was demonstrated using standard H&E-stained sections (Figure 17A).

[0180] Hippocampal thickness Images of H&E-stained sections were obtained using a standard virtual histology scan (EasyScan, Motic Microscopes, Schertz, TX, USA) at 40x magnification as previously described. Unbiased measurements of hippocampal thickness were obtained using the "Incremental Distances" plugin (Image-Pro Premier; Media Cybernetics, Warrendale, PA, USA) to measure the distance between the white matter border and the edge of the hippocampal formation (i.e., from the edge of the stratum oriens to the edge of the stratum lacunosum) using a vertical line in 10-μm steps, with an average of 3 mm per slide. 2 An average of 100 measurements was taken over an area of ​​.

[0181] Neurofibrillary tangles, neuritic plaques and ASC positive counts Using an extended depth of focus (EDF) virtual tissue scan (EasyScan, Motic Microscopes), which allows a series of Z-stack images to be converted into a single image, tissue was scanned at 1 mm depth. 2Unbiased cell counts of AT-8 pTau-positive neurons, ASC-positive cells, and Aβ clusters were determined in the CA1, CA2, CA3, subiculum, and adjacent DG using the Image-Pro Premier (Media Cybernetics) program. 2 Three contours were drawn from each region of interest (ROI), encompassing the average area of ​​the brain. ROIs were determined by cytoarchitecture as previously described and sampled from postmortem brains: CA1 was sampled from the anterior region of the DG, CA2 from the posterior region of the DG, and CA3 from the region adjacent to the opening of the DG. For each region, CA scans covered all layers.

[0182] The cell density of positively stained cells and Aβ clusters in all contours was quantified by investigators blinded to the case data. Histological scans were reviewed (by RV) to determine criteria to avoid duplicate counts (e.g., positive processes or Aβ clusters [>20 μm]). 2 ] and connected positive nuclei [>10 μm 2 In a pilot study, we confirmed that the number of labeled cells and nuclei was correctly counted using the aforementioned counting profile (using an Image J cell counter).

[0183] The number density was estimated by applying the following formula:

number

[0184] where N is the total number of cells or clusters per volume of brain region; ΣQ - is the number of cells counted; V is the volume of the region of interest per sampling frame.

[0185] Data analysis In this study, all variables between two groups (AD and control) were compared using Student's t-test. Data are presented as mean ± standard deviation (SD); significance was assumed at p < 0.05. All statistical analyses and plots were performed using GraphPad Prism 9.0 (GraphPad Software, San Diego, CA).

[0186] result Donors and AD pathology The experiments in this study were designed to determine the density and distribution of Aβ and pTau, as well as the expression of inflammatory proteins NLRP1, NLRP3, ASC, and caspase-1 in postmortem human brains with and without neuropathological changes of AD. The neuropathological scores and demographics of low- and intermediate-AD patients are shown in Tables 4 and 5 below, respectively.

[0187] [Table 9]

[0188] [Table 10]

[0189] Braak scores in the low AD group ranged from 0 to II; no AD neuropathological changes to low AD neuropathological changes. Neuropathological scores for neurofibrillary tangle distribution ranged from B0 to B1, with a mode of B1. Aβ diffuse plaque distribution scores, TAR phase, and A scores ranged from A0 to A2, with a mode of A1. Neuritic plaque density or CERAD scores ranged from C0 to C1, with a mode of C0. Causes of death included cardiovascular complications (14 donors), renal failure (1 donor), cancer (1 donor), and liver failure (1 donor).

[0190] Donors with intermediate AD had moderate neuropathological changes, with Braak scores ranging from III to VI. Neuropathological scores for neurofibrillary tangle distribution ranged from B2 to B3, with a mode of B2. Aβ diffuse plaque distribution scores, Thal stage, and A scores ranged from A1 to A3, with A2 as the mode. Neuritic plaque density or CERAD scores ranged from C1 to C3, with C2 as the mode. The intermediate AD group had similar comorbidities to the low AD group, including cardiovascular disease (10 donors), renal failure (4 donors), cancer (1 donor), and liver failure (1 donor). The age of low AD donors was not significantly higher than that of intermediate AD donors (p > 0.05). Furthermore, the two groups did not differ significantly in terms of PMI time (p > 0.05).

[0191] Hippocampal assessment The changes in cell density in the CA1 region may be due to the shrinkage of the layers due to changes in the neural disk. Furthermore, the volumetric reduction of the layers may also be related to differences in layer thickness. To address this possibility, we measured the hippocampal layers between the gray / white matter interface and the alveolar surface. When hippocampal thickness was measured in the CA1, CA2, and CA3 regions (Figure 17B), there was no significant difference in hippocampal thickness between intermediate and low AD cases (p>0.05), indicating that the changes in cell population are unrelated to changes in hippocampal volume.

[0192] The number of Aβ clusters in the hippocampus does not differ between intermediate and early AD Pathological changes, including histopathological assessment of Aβ deposition and staging of neurofibrillary changes, were also investigated. It was important to determine whether the amount of Aβ clusters varied between low and intermediate AD within our ROIs. In the CA region and the DG of the hippocampus, the number of Aβ clusters did not significantly change between intermediate AD cases compared with low AD (p > 0.05; Figures 18A, 18C, and 18E). However, in intermediate AD cases, a significant increase in Aβ deposition was observed in the subiculum (p = 0.02; intermediate AD: 54.08 ± 25.62, n = 17; low AD: 22.94 ± 20.75, n = 17) and entorhinal cortex (p = 0.01; intermediate AD: 71.27 ± 28.43, n = 17; low AD: 23.62 ± 24.46, n = 17). Outside the ROI, we examined the neuropathological scores (e.g., Thal score or A score) of diffuse Aβ plaques identified during case evaluation. The mean Thal score for intermediate AD cases was "A2" and the mean CERAD (e.g., neuritic amyloid plaque extent or C score) score was "C2," whereas the mean Thal score for low AD cases was "A1" and the mean CERAD score was "C0." The increase in Aβ plaques observed in the subiculum and entorhinal cortex indicates the presence of multiple regions susceptible to significant changes in protein accumulation as neurodegeneration progresses.

[0193] Tau positivity increased in CA1 in intermediate AD cases. pTau levels are used in the Braak staging system, which is based on the development of neurofibrillary tangles and neurites. We sought to determine whether the number of neurons with neurofibrillary tangles changes between intermediate and low AD. The number of pTau-positive neurons was significantly increased in intermediate AD cases compared with low AD cases in the subiculum (p=0.0006; intermediate AD: 64±48, n=17; low AD: 6.2±14, n=17) and entorhinal cortex (p<0.0001; intermediate AD: 55±36, n=17; low AD: 7.5±14, n=17). In the hippocampus, the intermediate AD group showed significant increases in the DG (p<0.04; intermediate AD: 39±27, n=17; low AD: 7.5±2.8, n=17), CA2 (p<0.002; intermediate AD: 99±56, n=17; low AD: 16±8, n=17), and CA1 regions (p<0.0002; intermediate AD: 99.96±57, n=17; low AD: 16.33±13, n=17), but not in CA3 (p>0.05; Figures 18B, 18D, and 18F). The increased number of neurofibrillary tangles illustrates the extent of neurodegeneration occurring in specific hippocampal regions. These data also suggest that the number of neurofibrillary tangles remains unchanged in certain brain regions (e.g., CA3), thus indicating that certain regions remain at this stage of AD pathology.

[0194] NLRP3 is primarily present in microglia-like structures Although the increase in pTau-positive neurons in intermediate AD cases has been well documented, the distribution and location of the sensor NLRP3 in AD cases has not been fully characterized. In early AD cases, NLRP3 is present in all regions of the hippocampus with a comparable distribution pattern in microglia (CA1: Figure 19A and CA2: Figure 19B). Similarly, in intermediate AD cases, NLRP3-positive microglial whorls were observed in both the CA1 (Figure 19C) and CA2 (Figure 19D) regions of the hippocampus, indicating that NLRP3 is present in microglia in two distinct regions of the hippocampus: CA1 and CA2.

[0195] Perinuclear expression of NLRP1 is present in hippocampal neurons of CA1 After confirming the morphological changes of NLRP3 in intermediate AD cases, we set out to detect changes in NLRP1 expression. In the intermediate AD cases, a small number of neurons in the CA1, but not the CA2, region showed perinuclear expression of NLRP1 (Figures 20A and 20B). In contrast, in the intermediate AD cases, more neuronal perinuclear expression was observed in the CA1 and CA2 hippocampal regions (Figures 20C and 20D), indicating differential expression of NLRP1 in neurons within the CA1 and CA2 regions of the hippocampus associated with AD pathology.

[0196] Detecting differential expression of ASC in neurons and microglia using antibodies raised against different epitopes of ASC Next, we evaluated whether intermediate AD brains exhibited altered expression of ASC in the hippocampus and entorhinal cortex. Mouse anti-ASC was found primarily in cells with microglia-like morphology in both the low AD and intermediate AD brains (Figures 21A and 21C). Furthermore, we observed a significant increase in the number of mouse anti-ASC-labeled microglia-like cells in all examined ROIs: EC (p = 0.0002; intermediate AD: 201 ± 87, n = 17; low AD: 86 ± 53, n = 17) and subiculum (p = 0.01; intermediate AD: 214 ± 105, n = 17; low AD: 74 ± 65, n = 17). DG (p = 0.001; mid-AD: 236 ± 87, n = 17; low-AD: 104 ± 53, n = 17), CA3 (p = 0.002; mid-AD: 214 ± 111, n = 17; low-AD: 105 ± 46, n = 17), CA2 (p = 0.002; mid-AD: 301 ± 111, n = 17; low-AD: 98 ± 46, n = 17), and CA1 (p = 0.015; mid-AD: 238 ± 162, n = 17; low-AD: 99 ± 66, n = 17; Figure S1E) all demonstrated upregulated microglial ASC protein expression in AD pathology.

[0197] Because ASC expression in human neurons during AD pathology is key to understanding how AD progresses, we sought to determine whether ASC expression occurs at different rates between the hippocampal formation and various regions of the entorhinal cortex (Fig. 21B and 21D). Therefore, using IC100, an antibody specific for human ASC, we detected increased ASC expression in neurons mainly in the DG (p = 0.002; mid-AD: 270 ± 189, n = 17; low-AD: 49 ± 35, n = 17), CA3 (p = 0.003; mid-AD: 260 ± 84, n = 17; low-AD: 39 ± 31, n = 17), CA2 (p = 0.001; mid-AD: 260 ± 93, n = 17; low-AD: 51 ± 42, n = 17), CA1 (p = 0.005; mid-AD: 219 ± 93, n = 17; low-AD: 72 ± 42, n = 17), and subiculum (p = 0.00; mid-AD: 267 ± 93, n = 17; low-AD: 86 ± 42, n = 17). Interestingly, in the EC, a region commonly present in early stages of AD pathology, there was no significant difference in the number of neurons detected with IC100 (p>0.05; Figure 21F), which may typically indicate a temporal response or indicate that the antibody specific for human ASC identifies a response specific to human neurons.

[0198] Caspase-1 expression is found in the parenchyma of low- and intermediate-stage AD cases Because the most significant pathological changes between intermediate AD cases and controls were evident between the CA1 and CA2 hippocampal regions, we further examined caspase-1 expression by focusing on these regions. In both low-AD (Figures 22A, 22B, and 22C) and intermediate-AD (Figures 22G, 22H, and 22I) cases, caspase-1 expression was present in the tissue parenchyma at the site of diffuse amyloid plaque-like formations and pTau neurofibrillary tangles in the CA1 region. Importantly, intermediate-AD cases (Figures 22J, 22K, and 22L) showed more prominent caspase-1 expression, amyloid plaque formation, and pTau neurofibrillary tangles in the CA2 region compared with low-AD cases (Figures 22D, 22E, and 22F), suggesting increased caspase-1 activity and consequent inflammasome activity in the hippocampus of intermediate-AD patients.

[0199] Consideration In this study, we used a panel of human-specific inflammasome antibodies to compare the expression of inflammasome signaling proteins NLRP1, NLRP3, ASC, and caspase-1 in the brains of donors with intermediate and low AD pathology. The results indicate that in early AD pathology, neurons and microglia exhibit increased expression of inflammasome proteins prior to cellular and hippocampal volume loss. NLRP1 was primarily expressed in neurons, whereas NLRP3 was present in microglia. Furthermore, caspase-1 was present in the hippocampal tissue parenchyma. Importantly, IC100 revealed increased ASC expression in neurons during the early stages of AD, whereas a commercially available antibody against a different domain of the ASC protein (the CARD domain) primarily labeled microglia. To our knowledge, this is the first demonstration of ASC expression in distinct cell populations during the early stages of AD, highlighting the importance of the inflammasome in early AD pathology.

[0200] Recent evidence has accumulated that inflammasome-inducing cytokines and inflammasome signaling proteins released from activated microglia interact with AD-related proteins, exacerbating the pathological progression and cellular damage of AD. NLRP3 expression in microglia from early and intermediate AD cases has been previously reported (see references). NLRP3 may contribute to chronic neuroinflammation through the production of IL-1β, resulting in impaired Aβ plaque clearance. In intermediate AD cases, NLRP3 expression was observed in microglia adjacent to neurons or present in clusters. The clustered distribution of NLRP3 may be mediated by TANK-binding kinase 1 (TBK1), which interacts with tau protein. Furthermore, we demonstrated that NLRP1 is expressed in the cytoplasm of hippocampal neurons and is upregulated in intermediate AD cases. This observation is consistent with our previous studies showing that NLRP1 is present in motor neurons in the anterior horn of the human spinal cord and is upregulated after spinal cord injury, traumatic brain injury, stroke, and brain aging. In addition, Saresella et al. and Yap et al. reported that NLRP1 is primarily expressed in hippocampal pyramidal neurons and is activated by aggregated Aβ. Recent studies have shown that knockout of NLRP1 in AD model mice reduces Aβ plaque burden, normalizes hippocampal dendritic spines, and improves spatial and episodic memory performance. Collectively, these results suggest that the NLRP1 inflammasome is involved in neurodegenerative processes in neurons, in addition to upregulating the NLRP3 inflammasome in microglia. However, it remains unclear whether other inflammasomes present in various CNS cell types also contribute to the enhanced inflammatory response in AD, and whether tau, Aβ, or ASC specks released from inflammasome-activated cells trigger cell death processes such as pyroptosis, which may contribute to hippocampal cell death in the early stages of AD.

[0201] Although the ages of the cohorts examined in this study were similar in the intermediate and early AD groups, cognitive impairment was higher in the intermediate AD group. We cannot exclude the possibility that some early AD cases would have progressed to AD if the donors had lived longer. Nevertheless, our data suggest that changes in inflammasome protein expression in neurons and microglia accompany AD neurodegeneration in the central nervous system of aging individuals.

[0202] The hippocampal formation cortical zone showed no difference in thickness between low and intermediate AD, indicating that neuronal death and atrophy were not evident in these cases. Two other studies examined hippocampal thickness and neuronal counts in AD cases classified into different Braak stages V-VI and reported hippocampal neuronal loss in Braak stages V-VI, supporting our findings. In those studies, the authors evaluated neuropathological changes in all CA regions of the hippocampus between low and intermediate AD groups and found no significant difference in the number of Aβ plaque clusters between the low and intermediate AD groups. However, we found that the plaque morphology was more dense in the low AD group than in the intermediate group. In contrast, in the intermediate group, the number of pTau-labeled neurons was significantly increased in the subiculum, CA1, CA2, and DG regions, consistent with previous studies of tau pathology in postmortem AD cases.

[0203] Protein aggregation is associated with over 30 proteinopathies, including many neurodegenerative diseases such as AD, where both Aβ and tau aggregates are present. Recent evidence indicates that heterotypic interactions of aggregation proteins occur in diverse amyloid processes, and these interactions alter fundamental aspects of amyloid aggregation, such as seeding, aggregation kinetics, and toxicity. In this study, we investigated the expression of three protein aggregates: ASC, Aβ, and p-tau. The expression of all three aggregation proteins is higher in intermediate AD compared with low AD, indicating increased deposition associated with disease progression. In AD pathology, ASC specks mediate Aβ aggregation in the extracellular space. 1-42It has been shown that Aβ cross-seeding occurs in microglia. 1-42 This suggests that disease-associated protein aggregation may regulate the morphology or aggregation rate of amyloidogenic proteins, enhancing the toxicity of AD. Supporting this idea are the observations that excessive inflammasome stimulation in microglia and neurons induces ASC oligomerization to form inflammasome complexes, and that this response is exacerbated by tauopathy. Our findings demonstrating the presence of ASC specks in the brains of AD patients suggest that immunotherapy targeting protein aggregates may be a promising target for the treatment of neurodegenerative diseases.

[0204] In addition to the canonical ASC, the inflammasome adaptor ASC has different isoforms. These other isoforms, termed ASCb, ASCc, and ASCd, differ in amino acid composition. ASC and ASCb are very similar, both containing PYD and CARD domains. However, the length of the interdomain linker differs. ASCc contains a CARD domain and a fragment of the PYD, whereas ASCd has a partial PYD and lacks the CARD domain. Full-length ASC and ASCb colocalize with the sensors NLRP-3 and procaspase-1 and activate inflammasomes. However, ASCb does not form typical ASC specks but rather forms filamentous aggregates of ASC, resulting in lower levels of inflammasome activation, as determined by IL-1β release. ASC can oligomerize more rapidly and assemble into more uniformly sized oligomers than ASCb. In contrast, ASCc only colocalizes with caspase-1 and reduces IL-1β release in the presence of ASC, potentially inhibiting inflammasomes. ASCd, whose function remains undefined, does not colocalize with NLRP3 and is unable to produce mature IL-1β. Different combinations of ASC splice variants may be expressed intracellularly, which may affect inflammasome activity at different stages of the inflammatory response. Our results show that antibodies raised against two distinct epitopes of ASC (CARD vs. PYD) differentially distinguish neurons from microglia in the early stages of AD, suggesting that ASC exists in different conformational states within different populations of CNS cells or that different ratios of ASC isoforms expressed in microglia and neurons are differentially recognized by the two antibodies. Furthermore, the expression of ASC in the early stages of AD pathology is consistent with our previous studies showing that ASC is elevated in the blood of patients with mild cognitive impairment and that ASC is a reliable biomarker for AD.Therefore, both ASC and IL-18 were significantly higher in the serum of MCI patients compared with controls, and ASC protein levels were also higher in the serum of MCI patients compared with AD patients.Furthermore, future studies are needed to understand the differential expression ratios of activating and inhibitory isoforms of ASC in cells and how they promote inflammation in the early stages of infection and tissue injury.

[0205] In this study, we compared the cell-specific staining properties of two ASC antibodies (one against the CARD of the ASC and the other against the PYD, IC100) with a commercially available antibody panel identifying Aβ, pTau, NLRP1, NLRP3, and caspase-1. Compared with a commercially available mouse ASC antibody, IC100 identified neurons at an early stage of neurodegeneration in the intermediate stages of AD. The expression pattern of IC100 was consistent with Aβ and pTau staining in postmortem AD brains, revealing the spatial and temporal relationship of this sensor molecule in the pathogenesis of AD neuropathology. The report of a differential pattern of IC100 neuronal immunostaining without evidence of cell loss in human AD specimens highlights the importance of targeting this unique ASC configuration for future molecular imaging and therapeutic approaches to reduce neuronal vulnerability and subsequent ASC speckle shedding, which leads to continued disease progression.

[0206] Several alternative markers are under development to assess early signs of disease development, including molecular neuroimaging, blood or CSF biomarkers, and sensitive indicators of cognitive impairment. Furthermore, numerous studies and potential therapeutic interventions are currently being investigated to prevent, mitigate, or reverse the effects of AD pathology. The inflammasome pathway provides an attractive therapeutic target for mitigating the harmful inflammatory secondary injury cascade. IC100 is a humanized monoclonal antibody against the adaptor protein ASC and has therapeutic benefit in treating several experimental models of neurodegeneration, injury, and aging. In this study, IC100 immunoreactivity demonstrates regional specificity and a cell-specific labeling pattern based on the severity of AD pathology, in contrast to commercially available mouse anti-ASC antibodies, which are primarily immunoreactive with cells with microglia-like morphology.

[0207] In conclusion, our observations of donor brains with low and intermediate AD pathology indicate that the NLRP1 inflammasome is primarily present in neurons, whereas the NLRP3 inflammasome is primarily present in microglia. Furthermore, ASC is present in both neurons and microglia. However, antibodies against the CARD of ASC detected ASC primarily in microglia, whereas IC100, which recognizes the PYD of ASC, detected it primarily in neurons. Thus, our data suggest that IC100 identifies neurons at an early stage of neurodegeneration in intermediate AD, and that ASC expression corresponds to the levels of Aβ and pTau in postmortem AD brains. Collectively, these results indicate that IC100 is a promising novel therapeutic approach for the early diagnosis and treatment of AD.

[0208] Numbered Embodiments of the Present Disclosure Other subject matter contemplated by the present disclosure is described in the following numbered embodiments: 1. A method for treating pulmonary inflammation in a patient in need thereof, said method comprising administering to said patient a composition comprising an agent that inhibits inflammasome signaling, thereby treating pulmonary inflammation.

[0209] 2. The method of embodiment 1, wherein said pulmonary inflammation is caused by a condition selected from a viral infection, a central nervous system (CNS) injury, a neurodegenerative disease, an autoimmune disease, asthma, chronic obstructive pulmonary disease, cystic fibrosis, interstitial lung disease, and acute respiratory distress syndrome.

[0210] 3. The method of embodiment 2, wherein said CNS injury is selected from traumatic brain injury (TBI), stroke, and spinal cord injury (SCI).

[0211] 4. The method of embodiment 2, wherein said neurodegenerative disease is selected from the group consisting of Alzheimer's disease (AD) / dementia, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), and Parkinson's disease (PD).

[0212] 5. The method of any one of embodiments 1 to 4, wherein administration of the composition results in inhibition of inflammasome activation in germ cells of the patient.

[0213] 6. The method of any one of embodiments 1 to 4, wherein administration of the composition results in a decrease in the level of caspase-1, nucleotide-binding leucine-rich repeat pyrin domain-containing protein 1 (NLRP1), nucleotide-binding leucine-rich repeat pyrin domain-containing protein 2 (NLRP2), nucleotide-binding leucine-rich repeat pyrin domain-containing protein 3 (NLRP3), NLR family CARD domain-containing protein 4 (NLRC4), caspase-11, X-linked inhibitor of apoptosis protein (XIAP), pannexin-1, caspase-activation recruitment domain-containing apoptosis-associated speck-like protein (ASC), interleukin-18 (TL-18), high-mobility group box 1 (HMGB1), or absent in melanoma 2 (AIM2) in lung cells of the patient compared to a control, and the control is an untreated patient.

[0214] 7. The method of embodiment 5 or 6, wherein said lung cells are type II pneumocytes.

[0215] 8. The method of any one of embodiments 1 to 5, wherein administration of the composition results in a reduction in acute lung injury (ALI) compared to a control, wherein the control is an untreated patient.

[0216] 9. The method of embodiment 8, wherein the alleviation of ALI is evidenced by a decrease in neutrophil infiltration into the alveoli and / or interstitial spaces, a reduction or elimination of alveolar septal thickening, or a combination thereof.

[0217] 10. The method of any one of embodiments 1 to 9, wherein the agent is an extracellular vesicle (EV) uptake inhibitor, an antibody that binds to an inflammasome component, or a combination thereof.

[0218] 11. The method of embodiment 10, wherein the EV uptake inhibitor is a compound or an antibody, and the antibody is selected from Table 1.

[0219] 12. The method of any one of embodiments 10-11, wherein the agent is an EV uptake inhibitor in combination with an antibody that binds to an inflammasome component.

[0220] 13. The method of embodiment 12, wherein the EV uptake inhibitor is heparin.

[0221] 14. The method of embodiment 13, wherein the heparin is enoxaparin.

[0222] 15. The method of any one of embodiments 10 to 14, wherein the antibody that binds to the inflammasome component is an antibody that specifically binds to a component of the mammalian AIM2, NLRP1, NLRP2, NLRP3, or NLRC4 inflammasome.

[0223] 16. The method of embodiment 10 or 15, wherein said inflammasome component is caspase-1, ASC or AIM2.

[0224] 17. The method of embodiment 16, wherein the inflammasome component is ASC.

[0225] 18. The method of embodiment 17, wherein the antibody binds to the N-terminal PYRIN-PAAD-DAPIN domain (PYD), the C-terminal caspase recruitment domain (CARD) of the ASC protein, or an epitope derived from the PYD or CARD domain.

[0226] 19. The method of embodiment 17, wherein said antibody binds to an amino acid having at least 85% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:2.

[0227] 20. The method of any one of embodiments 17-19, wherein said antibody inhibits ASC activity in the lungs of said patient.

[0228] 21. The method of any one of embodiments 1-20, wherein the composition is formulated together with a pharmaceutically acceptable carrier or diluent.

[0229] 22. The method of any one of embodiments 1-21, wherein the composition is administered intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation.

[0230] 23. A method for treating inflammation in the lungs of a patient suffering from a viral infection or central nervous system (CNS) injury, the method comprising administering to the patient a composition comprising an agent that inhibits inflammasome signaling, thereby treating inflammation in the lungs of the patient.

[0231] 24. The method of embodiment 23, wherein said CNS injury is selected from the group consisting of traumatic brain injury (TBI), stroke, and spinal cord injury (SCI).

[0232] 25. The method of any one of embodiments 23-24, wherein administration of the composition results in inhibition of inflammasome activation in lung cells of the patient.

[0233] 26. The method of any one of embodiments 23-24, wherein administration of the composition results in a decrease in the level of caspase-1, NLRP1, NLRP2, NLRP3, NLRC4, caspase-11, XIAP, pannexin-1, apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC), interleukin-18 (IL-18), high-mobility group box 1 (HMGB1), or absent in melanoma 2 (AIM2) in the patient's germ cells compared to a control, and the control is an untreated patient.

[0234] 27. The method of embodiment 25 or 26, wherein said lung cells are type II pneumocytes.

[0235] 28. The method of any one of embodiments 23-27, wherein administration of said composition results in a reduction in acute lung injury (ALI) compared to a control, wherein the control is an untreated patient.

[0236] 29. The method of embodiment 28, wherein said reduction in ALI is evidenced by a decrease in neutrophil infiltration into the alveoli and / or interstitial spaces, a decrease or disappearance of alveolar septal thickening, or a combination thereof.

[0237] 30. The method of any one of embodiments 23 to 29, wherein the agent is an extracellular vesicle (EV) uptake inhibitor, an antibody that binds to an inflammasome component, or a combination thereof.

[0238] 31. The method of embodiment 30, wherein the EV uptake inhibitor is a compound or an antibody, and the antibody is selected from Table 1.

[0239] 32. The method of any one of embodiments 30-31, wherein the agent is an EV uptake inhibitor in combination with an antibody that binds to an inflammasome component.

[0240] 33. The method of embodiment 32, wherein the EV uptake inhibitor is heparin.

[0241] 34. The method of embodiment 33, wherein the heparin is enoxaparin.

[0242] 35. The method of any one of embodiments 30 to 34, wherein the antibody that binds to an inflammasome component is an antibody that specifically binds to a component of the mammalian AIM2, NLRP1, NLRP2, NLRP3, or NLRC4 inflammasome.

[0243] 36. The method of embodiment 30 or 35, wherein the inflammasome component is caspase-1, ASC, or AIM2.

[0244] 37. The method of embodiment 36, wherein the inflammasome component is ASC.

[0245] 38. The method of embodiment 37, wherein the antibody binds to the PYD, CARD domain, or an epitope derived from the PYD or CARD domain of the ASC protein.

[0246] 39. The method of embodiment 37, wherein said antibody binds to an amino acid having at least 85% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:2.

[0247] 40. The method of any one of embodiments 37-39, wherein the antibody inhibits ASC activity in the lungs of the patient.

[0248] 41. The method of any one of embodiments 23-40, wherein the composition is formulated together with a pharmaceutically acceptable carrier or diluent.

[0249] 42. The method of any one of embodiments 23-41, wherein the composition is administered intracerebroventricularly, intraperitoneally, intravenously, intranasally, or by inhalation.

[0250] 43. A monoclonal antibody or antibody fragment thereof that binds to an apoptosis-associated speck-like protein containing a caspase-activating recruitment domain (ASC), wherein the antibody or antibody fragment specifically binds to an epitope of ASC, and the epitope comprises or consists of the amino acid sequence of SEQ ID NO: 5, or 5 to 10, 10 to 15, or 15 to 20 amino acids of SEQ ID NO: 5.

[0251] 44. A monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a heavy chain variable (VH) region and a light chain variable (VL) region, and the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and HCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3.

[0252] 45. A monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a light chain variable (VL) region and a heavy chain variable (VH) region, and the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3.

[0253] 46. ​​A monoclonal antibody or antibody fragment thereof that specifically binds to ASC, wherein the antibody or antibody fragment comprises a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 6, HCDR2 of SEQ ID NO: 7, and LCDR3 of SEQ ID NO: 8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3; and the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3.

[0254] 47. The monoclonal antibody or antibody fragment thereof of embodiment 44, wherein the VH region amino acid sequence comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22.

[0255] 48. The monoclonal antibody or antibody fragment thereof of embodiment 45, wherein the VL region amino acid sequence comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

[0256] 49. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21 or 22; and the VL region amino acid sequence comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

[0257] 50. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 18 or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 28 or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 28.

[0258] 51. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 18 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 29 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29.

[0259] 52. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 18 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 30 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30.

[0260] 53. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 18 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 18; and the VL region amino acid sequence comprises SEQ ID NO: 31 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31.

[0261] 54. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 19 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 28 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28.

[0262] 55. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 19 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 29 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29.

[0263] 56. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 19 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 30 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30.

[0264] 57. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 19 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 19; and the VL region amino acid sequence comprises SEQ ID NO: 31 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31.

[0265] 58. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 20 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 20; and the VL region amino acid sequence comprises SEQ ID NO: 28 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28.

[0266] 59. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 20 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 20; and the VL region amino acid sequence comprises SEQ ID NO: 29 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29.

[0267] 60. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 20 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 20; and the VL region amino acid sequence comprises SEQ ID NO: 30 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30.

[0268] 61. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 20 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 20; and the VL region amino acid sequence comprises SEQ ID NO: 31 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31.

[0269] 62. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 21 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 21; and the VL region amino acid sequence comprises SEQ ID NO: 28 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28.

[0270] 63. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 21 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 21; and the VL region amino acid sequence comprises SEQ ID NO: 29 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29.

[0271] 64. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 21 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 21; and the VL region amino acid sequence comprises SEQ ID NO: 30 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30.

[0272] 65. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 21 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 21; and the VL region amino acid sequence comprises SEQ ID NO: 31 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31.

[0273] 66. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 22 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the VL region amino acid sequence comprises SEQ ID NO: 28 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28.

[0274] 67. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 22 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the VL region amino acid sequence comprises SEQ ID NO: 29 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 29.

[0275] 68. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 22 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the VL region amino acid sequence comprises SEQ ID NO: 30 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 30.

[0276] 69. The monoclonal antibody or antibody fragment thereof of embodiment 46, wherein the VH region amino acid sequence comprises SEQ ID NO: 22 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 22; and the VL region amino acid sequence comprises SEQ ID NO: 31 or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 31.

[0277] 70. The monoclonal antibody or antibody fragment thereof according to any one of embodiments 44 to 69, wherein said ASC is a human ASC protein.

[0278] 71. The monoclonal antibody fragment of any one of embodiments 44 to 70, wherein the antibody fragment is a Fab, F(ab')2, Fab', scFv, single domain antibody, diabody or single chain camelid antibody.

[0279] 72. The monoclonal antibody or antibody fragment thereof according to any one of embodiments 44 to 71, wherein said monoclonal antibody or antibody fragment thereof is a human, humanized or chimeric antibody or antibody fragment thereof.

[0280] 73. An isolated nucleic acid molecule encoding a monoclonal antibody or antibody fragment thereof according to any one of embodiments 44 to 72.

[0281] 74. An expression vector comprising the nucleic acid molecule of embodiment 73.

[0282] 75. The expression vector of embodiment 32, wherein the nucleic acid molecule is operably linked to a regulatory sequence suitable for expression of the nucleic acid segment in a host cell.

[0283] 76. A recombinant host cell comprising an expression vector according to embodiment 74 or 75.

[0284] 77. A method for producing an antibody or antibody fragment that specifically binds to ASC, comprising culturing a recombinant host cell comprising the expression vector of embodiment 74 or 75 under conditions in which the nucleic acid molecule is expressed, thereby producing the monoclonal antibody or antibody fragment thereof that specifically binds to ASC.

[0285] 78. A pharmaceutical composition comprising a monoclonal antibody or antibody fragment thereof according to any one of embodiments 44 to 72, and a pharmaceutically acceptable carrier, diluent or excipient.

[0286] 79. A method for treating inflammation in a subject, comprising administering to the subject a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof described in any one of embodiments 44 to 72, thereby treating inflammation in the subject.

[0287] 80. The method of embodiment 79, wherein administration of said monoclonal antibody or antibody fragment thereof reduces the level of at least inflammatory cytokines.

[0288] 81. The method of embodiment 80, wherein the inflammation is inflammasome-associated inflammation.

[0289] 82. The method of embodiment 81, wherein said inflammasome-associated inflammation is associated with a viral infection, a central nervous system (CNS) injury, an autoimmune disease, or a neurodegenerative disease.

[0290] 83. The method of embodiment 82, wherein said CNS injury is selected from the group consisting of traumatic brain injury (TBI), stroke, and spinal cord injury (SCI).

[0291] 84. The method of embodiment 82, wherein said autoimmune disease or neurodegenerative disease is amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, muscular dystrophy (MD) or multiple sclerosis (MS).

[0292] 85. The method of any one of embodiments 79 to 84, wherein administration of the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject.

[0293] 86. The method of any one of embodiments 79 to 84, wherein administration of the monoclonal antibody or antibody fragment thereof results in a decrease in the activity of ASC compared to a control.

[0294] 87. The method of embodiment 86, wherein the control is an untreated subject.

[0295] 88. The method of any one of embodiments 79 to 87, wherein said administration is performed intracerebroventricularly, intraperitoneally, intravenously or by inhalation.

[0296] 89. A method for treating multiple sclerosis (MS) in a subject, comprising administering to the subject a therapeutically effective amount of a monoclonal antibody or antibody fragment thereof described in any one of embodiments 44 to 72, thereby treating MS in the subject.

[0297] 90. The method of embodiment 89, wherein administration of said monoclonal antibody or antibody fragment thereof reduces the level of at least inflammatory cytokines.

[0298] 91. The method of embodiment 89, wherein administration of the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject.

[0299] 92. The method of any one of embodiments 89 to 91, wherein administration of the monoclonal antibody or antibody fragment thereof results in a decrease in the activity of ASC compared to a control.

[0300] 93. The method of embodiment 92, wherein the control is an untreated subject.

[0301] 94. The method of any one of embodiments 89-93, wherein said administration is intracerebroventricular, intraperitoneal, intravenous, intranasal, or by inhalation.

[0302] 95. The method of any one of embodiments 2, 23, or 82, wherein the viral infection is caused by a coronavirus or an influenza virus.

[0303] 96. The method of embodiment 95, wherein the coronavirus is a SARS virus or a MERS virus.

[0304] 97. The method of embodiment 96, wherein the SARS coronavirus is SARS-CoV or SARS-CoV-2.

[0305] 98. The method of embodiment 95, wherein the influenza virus is pandemic influenza.

[0306] 99. The method of embodiment 98, wherein the novel influenza virus is influenza A H5N1 (avian influenza) or influenza A H1N1 (swine influenza).

[0307] 100. The method of embodiment 95, wherein the viral infection causes acute respiratory distress syndrome (ARDS).

[0308] This application contains a Sequence Listing, which is incorporated herein by reference in its entirety and is reproduced herein as follows:

[0309] Sequence Listing [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

change

change

change

change

change

change

change

change

change

change

change

change

change

change

change

[0310] The various embodiments described above can be combined to provide further embodiments. All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent literature referenced herein are incorporated herein by reference in their entirety. Aspects of the embodiments can be modified as necessary to employ concepts from the various patents, applications, and publications to provide further embodiments.

[0311] These and other changes can be made to the embodiments in light of the above detailed description. Generally, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and claims, but should be construed to include all possible embodiments, along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by this disclosure.

Claims

1. A method for treating an inflammatory neurological disease, the method comprising administering a therapeutically effective amount of an inflammasome antibody, wherein the antibody is a monoclonal antibody, i.e., IC100, that specifically binds to ASC, or an antibody fragment thereof.

2. 2. The method of claim 1, wherein the inflammatory neurological disease is selected from the group consisting of mild cognitive impairment (MCI), Alzheimer's disease (AD), Parkinson's disease, multiple sclerosis (MS), and amyotrophic lateral sclerosis (ALS).

3. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence comprises an HCDR1 of SEQ ID NO:6, an HCDR2 of SEQ ID NO:7, and an HCDR3 of SEQ ID NO:8, or a variant thereof having at least one amino acid substitution in HCDR1, HCDR2, and / or HCDR3; and The method of claim 1, wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 12, LCDR2 of SEQ ID NO: 13, and LCDR3 of SEQ ID NO: 14, or a variant thereof having at least one amino acid substitution in LCDR1, LCDR2, and / or LCDR3.

4. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence which is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

5. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 18, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

6. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO:19, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:19; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

7. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO:20, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:20; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

8. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO:21, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:21; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

9. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO:22, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:22; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 28, 29, 30, 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28, 29, 30 or 31.

10. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence which is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22; 2. The method of claim 1, wherein the VL region amino acid sequence comprises SEQ ID NO:28, or an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:

28.

11. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence which is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO:29, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:

29.

12. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence which is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 30, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:

30.

13. The IC100 monoclonal antibody or antibody fragment thereof comprises a heavy chain variable (VH) region and a light chain variable (VL) region; the VH region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 18, 19, 20, 21, 22, or an amino acid sequence which is at least 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 18, 19, 20, 21, or 22; The method of claim 1, wherein the VL region amino acid sequence of the monoclonal antibody or antibody fragment thereof comprises SEQ ID NO: 31, or an amino acid sequence that is at least 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:

31.

14. The method of claim 1 , wherein the ASC is a human ASC protein.

15. The antibody fragments include Fab, F(ab') ２ , Fab', scFv, single domain antibody, diabody or single chain camelid antibody.

16. The method of claim 1 , wherein the monoclonal antibody or antibody fragment thereof is a human, humanized, or chimeric antibody or antibody fragment thereof.

17. The method of claim 1, wherein administration of the monoclonal antibody or antibody fragment thereof reduces the level of at least a pro-inflammatory cytokine.

18. The method of claim 1, wherein administration of the monoclonal antibody or antibody fragment thereof results in inhibition of inflammasome activation in the subject.

19. 2. The method of claim 1, wherein administration of the monoclonal antibody or antibody fragment thereof results in a decrease in ASC activity compared to an untreated control subject.

20. 10. The method of claim 1, wherein the administration is intracerebroventricular, intraperitoneal, intravenous, or by inhalation.

21. A composition for treating an inflammatory neurological disease, the composition comprising a therapeutically effective amount of an inflammasome antibody, the antibody being a monoclonal antibody, i.e., IC100, or an antibody fragment thereof, that specifically binds to ASC.

22. 22. The composition of claim 21, wherein the inflammatory neurological disease is Alzheimer's disease.

23. 1. A method for imaging neurodegenerative neurons in an early stage Alzheimer's disease patient, the method comprising: providing a monoclonal antibody or antibody fragment thereof having an IC100 that specifically binds to ASC; Binding the antibody to ASCs in brain tissue; and Determining levels of IC100-ASC binding correlate with early Alzheimer's disease A method comprising:

Images