Methods for treating viral infections

Abstract

To provide methods and compositions for reducing virus titer and eliminating virus-infected cells from an individual.SOLUTION: Provided is the use of an anti-CD24 antibody in the production of medicament for use in a method for reducing hepatitis virus in an individual, where the method comprises: contacting the individual to a therapeutically effective dose of the anti-CD24 antibody for a period of time sufficient to reduce the amount of hepatitis virus, where the hepatitis virus is one or more of hepatitis B virus (HBV) and hepatitis delta virus (HDV).SELECTED DRAWING: None

Description

[Background technology]

[0001] Viral infections are a major cause of morbidity and mortality worldwide. Current treatments for many of these viruses are inadequate or nonexistent. For example, hepatitis B virus (HBV) infects over 300 million people worldwide, despite the existence of an effective vaccine.

[0002] Furthermore, current treatments for HBV fail to completely eliminate the virus, particularly from the liver where it integrates into the human genome, and such integrated, persistent HBV infection is a major risk factor for chronic hepatitis, cirrhosis, liver cancer, and premature death.

[0003] At present, current treatments are largely unable to eradicate chronic HBV infection. The goal of eliminating the HBV surface antigen (HBsAg), which serves to inhibit the host's immune response to HBV, is rarely achieved, and even then, occurs only after years of treatment. Treatment modalities are of great interest. Summary of the Invention

[0004] Methods are provided for significantly reducing and, in some cases, eliminating virally infected cells in an individual by administering a therapeutically effective dose of an anti-CD24 agent to the individual. This is for a period of time sufficient to substantially reduce the amount of virus and, in some cases, to eliminate virally infected cells in the individual. In some embodiments, the virus is a virus that causes chronic infection, for example, by integrating into the host genome. In some embodiments, the virus is a chronic hepatitis virus, such as hepatitis B, hepatitis C, or hepatitis D. In some embodiments, the virus is a lentivirus, such as HIV-1 or HIV-2. In some embodiments, the virus is a human papillomavirus (HPV). In some embodiments, the virus is a herpesvirus, such as herpes simplex virus (HSV), human cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), HHV-8, etc.

[0005] Anti-CD24 agents may specifically bind to CD24 and block its activity, for example, by blocking binding to one of its ligands (e.g., P-selectin, siglec10, etc.). Agents that specifically bind to CD24 include, but are not limited to, antibodies and fragments derived therefrom, non-antibody binding proteins such as adnectins and anticalins, high-affinity binding peptides, high-affinity binding nucleic acids, small molecules, etc. CD24 expression can also be reduced with CRISPR, siRNA, or locked nucleic acids (LNA) that target CD24. Alternatively, anti-CD24 agents may specifically bind to and block CD24's ligand. Binding of CD24 to Siglec-10, present on B cells, dendritic cells (DCs), macrophages, and neutrophils, leads to attenuation of the immune response, a "leave-me-alone" or "don't kill me" signal. Upregulation of CD24 by HBV and other chronic viral infections may allow infected cells to evade the immune response, leading to viral persistence and, in some cases, subsequent cancer formation.

[0006] It is shown herein that doses of anti-CD24 antibodies above about 8 mg / kg can result in rapid, overt clinical toxicity and subsequent death. Surprisingly, much lower ranges of anti-CD24 antibody doses significantly reduce the presence of chronic viruses, including, but not limited to, HBV, under conditions that do not cause substantial toxicity to uninfected cells, and in some cases, In some embodiments, treatment results in undetectable levels of viral antigen, e.g., HBsAg, in the blood of infected individuals. In some embodiments, levels of circulating viral antigen, e.g., HBsAg, are reduced by at least one log compared to baseline pre-treatment levels, and may be reduced by at least one log, at least two logs, at least three logs, or more. In some embodiments, individuals achieve seroconversion to an anti-HBsAg-positive phenotype after treatment. In some embodiments, individuals are tested for levels of viral antigen and / or anti-viral antibodies after treatment. These data indicate that there may be unacceptable toxicity with the therapeutic use of anti-CD24 antibodies at doses greater than about 8 mg / kg.

[0007] The effect on non-infected cells, such as hepatocytes, can be monitored by the release of hepatocyte markers into the blood of non-infected individuals exposed to the drug. Low levels of the markers are expected to be detected due to the death of infected cells in infected patients, but the dose is expected to be low enough so that there is no dose-limiting toxicity and any increase in the marker levels is transient. Adverse effects are preferably at a level of less than 3, and may be less than 2 or even less than 1, using conventional criteria, such as CDCAE v. 5. Importantly, not all hepatocytes in HBV-infected individuals are infected with HBV, and the liver can be repopulated with non-infected cells. Therefore, selective elimination of HBV-infected cells can have significant clinical benefits. In some embodiments, individuals are tested for adverse effects after treatment.

[0008] Advantages of the present method may include, for example, rapid and controlled loss of circulating viral antigens, killing and elimination of infected cells, a short course of treatment, and a convenient administration schedule. For example, the course of treatment may be less than 24 weeks, less than about 12 weeks, less than about 8 weeks, or less than about 4 weeks, such as 1 to 12 weeks, 2 to 12 weeks, 4 to 12 weeks, or 4 to 8 weeks. Administration may be weekly, twice weekly, every other day, daily, every two weeks, or the like, and in some embodiments, once weekly or once every two or four weeks. In some embodiments, two or more courses of treatment are administered.

[0009] In some embodiments, the anti-CD24 agent is an antibody specific for human CD24, which optionally is a humanized or fully human monoclonal antibody. In some embodiments, the anti-CD24 antibody is administered at a dose of less than 8 mg / kg body weight, less than 2.5 mg / kg body weight, less than 1 mg / kg body weight, less than 0.75 mg / kg body weight, less than 0.5 mg / kg body weight, less than 0.25 mg / kg body weight, less than 0.1 mg / kg body weight, less than 0.05 mg / kg body weight, or less than 0.01 mg / kg body weight. The therapeutic dose may be, for example, 0.1 to 5 mg / kg, 0.25 to 5 mg / kg, 0.5 to 5 mg / kg, 0.75 to 5 mg / kg, 1 to 5 mg / kg, or 0.1 to 2.5 mg / kg, 0.25 to 2.5 mg / kg, 0.5 to 2.5 mg / kg, 0.7 to 2.5 mg / kg, 0.1 to 1 mg / kg, 0.25 to 1 mg / kg, 0.5 to 1 mg / kg, 0.75 to 1 mg / kg, etc.

[0010] Administration of an anti-CD24 agent may be combined with the simultaneous administration of an agent that prevents reinfection of new cells. Such agents may include, for example, the entry inhibitor myrcludex-b, anti-NTBC antibodies, HBV nucleoside analogs (e.g., TDF, TAF, etc.), etc. Administration of an anti-CD24 agent may also be combined with a second antiviral agent, for example, an HBsAg release inhibitor (nucleic acid polymer), an HBV core inhibitor, an HBV (or HDV)-targeting siRNA, an immunomodulator (a TLR agonist, interferon including alpha or lambda, etc.), an RT or polymerase inhibitor, a prenylation inhibitor, a therapeutic vaccine, etc.

[0011] In other embodiments, administration of an anti-CD24 agent is combined with administration of an agent that specifically targets a second antigen upregulated on virally infected cells. Markers of interest include CD1 Examples of antigens that bind to the second antigen include, but are not limited to, CD5, CD104, CD257, CD105, CD133, and CD47. In some embodiments, the agent that binds to the second antigen is an antibody. In some embodiments, a bispecific antibody that targets CD24 and the second antigen is administered.

[0012] In other embodiments, methods are provided for significantly reducing, and in some cases eliminating, virally infected cells in an individual by administering to the individual a therapeutically effective dose of an agent selected from an anti-CD15 agent, an anti-CD104 agent, an anti-CD133 agent, an anti-CD257 agent, or a combination thereof for a period sufficient to substantially eliminate virally infected cells. In some embodiments, the agent is other than an anti-CD47 agent. In some embodiments, the virus is a virus that causes chronic infection, for example, by integrating into the host genome. In some embodiments, the virus is a chronic hepatitis virus, e.g., hepatitis B, hepatitis C, or hepatitis D. In some embodiments, the virus is a lentivirus, e.g., HIV-1 or HIV-2. In some embodiments, the virus is a human papillomavirus (HPV). In some embodiments, the virus is a herpesvirus, e.g., herpes simplex virus (HSV), human cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), HHV-8, etc. Agents that specifically bind to CD15, CD104, CD133, CD47, and CD257 include, but are not limited to, antibodies and fragments derived therefrom, non-antibody binding proteins such as adnectins and anticalins, high affinity binding peptides, high affinity binding nucleic acids, small molecules, and the like.

[0013] In some embodiments, the agent is an antibody.Anti-CD15 agents can specifically bind to CD15 and block its activity, for example, by blocking binding to one of its ligands, such as P-selectin, CD43, CD44, etc.The expression of CD15, CD24, CD47, CD104, CD105, CD133, and CD257 can also be reduced with any antisense oligonucleotide, with or without CRISPR, siRNA, or locked nucleic acid (LNA), or chemically modified nucleotides / nucleosides that target them.CD15 expression can be further reduced by agents that target its host synthetic enzymes, such as fucosyltransferases FUT3, FUT5, FUT6, FUT7, and / or sialyltransferases ST3GAL3, ST3GAL4, and ST3GAL6.

[0014] In some embodiments, an antibody that binds to a marker selected from CD15, CD104, CD257, CD105, CD133, and CD47 and blocks binding of the marker to one of its ligands is administered at a dose of less than 40 mg / kg body weight, less than 35 mg / kg body weight, less than 30 mg / kg body weight, less than 25 mg / kg body weight, less than 20 mg / kg body weight, less than 15 mg / kg body weight, less than 10 mg / kg body weight, less than 5 mg / kg body weight, less than 2.5 mg / kg body weight, less than 1 mg / kg body weight, less than 0.75 mg / kg body weight, less than 0.5 mg / kg body weight, less than 0.25 mg / kg body weight, less than 0.1 mg / kg body weight, less than 0.05 mg / kg body weight, or less than 0.01 mg / kg body weight. The therapeutic dose may be, for example, 0.1 to 5 mg / kg, 0.25 to 5 mg / kg, 0.5 to 5 mg / kg, 0.75 to 5 mg / kg, 1 to 5 mg / kg, or 0.1 to 2.5 mg / kg, 0.25 to 2.5 mg / kg, 0.5 to 2.5 mg / kg, 0.7 to 2.5 mg / kg, 0.1 to 1 mg / kg, 0.25 to 1 mg / kg, 0.5 to 1 mg / kg, 0.75 to 1 mg / kg, 1 to 5 mg / kg, 5 to 10 mg / kg, 10 to 20 mg / kg, 20 to 30 mg / kg, 30 to 40 mg / kg, etc.

[0015] In other embodiments, compositions, e.g., anti-CD24 agents, for use in any of the methods described herein are provided, in effective unit doses, either alone or in combination with a second active agent as described above. In some embodiments, an anti-CD15 agent, an anti-CD104 agent, an anti-CD133 agent, an anti-CD47 agent, an anti-CD257 agent, or a combination thereof is provided in an effective unit use for use in the methods described herein. [Brief explanation of the drawings]

[0016] The invention will be best understood from the following detailed description when read in conjunction with the accompanying drawings. It is emphasized that, according to common practice, the various features of the drawings are not to scale. Conversely, the dimensions of the various features have been arbitrarily expanded or reduced for clarity. The drawings include the following figures:

[0017] [Figure 1] Anti-CD24 and anti-CD15 have been shown to prevent tumor formation in HBV-infected cells. [Figure 2-1]Monoclonal antibody-mediated phagocytic clearance of an HBV cell line (HepG2.2.15) is shown. HepG2.2.15 cells were labeled with mCherry (red) by transducing a lentiviral vector containing the mCherry gene under the PGK promoter. RAW 264.7 cells were cultured in IMDM medium with 10% FBS and activated with 50 nM phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) and 10 ng / ml M-CSF. After 2 days of activation, Raw264.7 cells were detached and labeled with 0.1 mM calcein AM (green) for 30 minutes at 37°C. Labeled HepG2.2.15 and Raw264.7 cells were mixed at a 1:10 ratio and seeded onto an 8-well chamber slide. They were then rapidly attached by low-speed centrifugation for 5 minutes. Cells were then cultured in IMDM containing 50 mM PMA and 10 ng / ml pigment epithelium-derived factor (PEDF). The indicated antibodies were added to each well to a final concentration of 10–20 μg / mL. After 24 h of co-culture, cells were washed three times with PBS and mounted in antifade medium containing DAPI. Images were captured using a KEYENCE BZ-X710 All-in-One Fluorescence Microscope (KEYENCE Corp. of USA, Itasca, IL, USA). The antibodies in each panel are: A. IgG2a negative control, B. anti-CD24, C. anti-CD15, D. anti-CD47, E. anti-CD104, F. anti-CD133, and G. anti-CD257. [Figure 2-2]Monoclonal antibody-mediated phagocytic clearance of an HBV cell line (HepG2.2.15) is shown. HepG2.2.15 cells were labeled with mCherry (red) by transducing a lentiviral vector containing the mCherry gene under the PGK promoter. RAW 264.7 cells were cultured in IMDM medium with 10% FBS and activated with 50 nM phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) and 10 ng / ml M-CSF. After 2 days of activation, Raw264.7 cells were detached and labeled with 0.1 mM calcein AM (green) for 30 minutes at 37°C. Labeled HepG2.2.15 and Raw264.7 cells were mixed at a 1:10 ratio and seeded onto an 8-well chamber slide. They were then rapidly attached by low-speed centrifugation for 5 minutes. Cells were then cultured in IMDM containing 50 mM PMA and 10 ng / ml pigment epithelium-derived factor (PEDF). The indicated antibodies were added to each well to a final concentration of 10–20 μg / mL. After 24 h of co-culture, cells were washed three times with PBS and mounted in antifade medium containing DAPI. Images were captured using a KEYENCE BZ-X710 All-in-One Fluorescence Microscope (KEYENCE Corp. of USA, Itasca, IL, USA). The antibodies in each panel are: A. IgG2a negative control, B. anti-CD24, C. anti-CD15, D. anti-CD47, E. anti-CD104, F. anti-CD133, and G. anti-CD257. DETAILED DESCRIPTION OF THE INVENTION

[0018] It is shown herein that HBV infection is associated with increased expression of host cell CD24 on the cell surface. Treatment of an in vivo model of HBV-infected human liver with a monoclonal antibody against human CD24 resulted in a rapid decline in HBV titers and a significant reduction, and in some cases, elimination, of HBV surface antigen (HBsAg). Current therapies can suppress extrachromosomal replication of HBV, but leave HBV-infected cells intact, with the integrated genome producing large amounts of immunosuppressive HBsAg expression. In some cases, treatment with anti-CD24 resulted in the death of HBV-infected cells, which eliminated all traces of HBV infection, eliminated infection-promoting integrated HBV, and eliminated immunosuppressive HBsAg. In some cases, this method actually eliminated HBV-infected cells with minimal damage to uninfected liver cells.

[0019] Other markers that also affect phagocytosis of virus-infected cells include CD15, CD104, CD133, CD47, and CD257. Blocking these markers on the surface of infected cells increases phagocytosis.

[0020] Before the present methods and compositions are described, it is to be understood that this invention is not limited to the particular method or compositions described, as such may, of course, vary. Further, the scope of the present invention will be limited only by the appended claims, and therefore all references thereto are to the terms "method" and "composition" as used herein. It is also to be understood that the terminology used is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0021] Where a range of values ​​is provided, unless the context clearly dictates otherwise, it is understood that each intervening value between the upper and lower limit of that range, to the tenth of the unit of the lower limit, is also specifically disclosed. Each smaller range between any stated value or intervening value within a stated range and any other stated value or intervening value within that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded, and each range in which either, neither, or both limits are included in the smaller range is also encompassed within the invention, subject to any specifically excluded limits in the stated range. Where a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also encompassed within the invention.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.Although any method and material similar or equivalent to the methods and materials described herein can be used to implement or test this invention, some potential and preferred methods and materials are described herein.All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publication is cited.In the event of any discrepancy, it should be understood that the present disclosure shall prevail over any disclosure of the incorporated publication.

[0023] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features which may be readily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0024] It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to "a cell" includes a plurality of such cells, and a reference to "the peptide" includes one or more peptides and equivalents thereof, such as polypeptides known to those skilled in the art.

[0025] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates, which may need to be independently confirmed.

[0026] The following description utilizes several terms commonly used in the art. The following definitions are provided to provide a clear and consistent understanding of the specification and claims, and the scope to be given such terms.

[0027] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms also apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.

[0028] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as amino acids that are later modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as naturally occurring amino acids, i.e., an alpha carbon bonded to a hydrogen, a carboxyl group, an amino group, and an R group, such as homoserine, norleucine, methionine sulfoxide, or methionine sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimetics refer to compounds that have a structure different from the general chemical structure of amino acids but function similarly to naturally occurring amino acids.

[0029] The terms "recipient," "individual," "subject," "host," and "patient" are used interchangeably herein and refer to any mammalian subject, particularly humans, for whom diagnosis, treatment, or therapy is desired. "Mammal" for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sport, or pet animals such as dogs, horses, cats, cows, sheep, goats, pigs, etc. In some embodiments, the mammal is a human.

[0030] The term "sample" encompasses, with respect to a patient, blood and other liquid samples of biological origin, solid tissue samples such as biopsy specimens or tissue cultures or cells derived therefrom, and the progeny thereof. This definition also includes samples that have been manipulated in any way after procurement, such as treatment with reagents, washing, or enrichment for particular cell populations. This definition also includes samples enriched for particular types of molecules, e.g., nucleic acids, polypeptides, etc.

[0031] The term "biological sample" encompasses clinical samples, including tissue obtained by surgical resection, tissue obtained by biopsy, cells in culture, cell supernatants, cell lysates, tissue samples, organs, bone marrow, blood, plasma, serum, aspirates, and the like. A "biological sample" includes a sample containing, or suspected of containing, target cells and / or normal control cells. The definition includes biological fluids derived therefrom (e.g., infected cells, etc.), such as samples containing polynucleotides and / or polypeptides obtained from such cells (e.g., cell lysates or other cell extracts containing polynucleotides and / or polypeptides). Biological samples containing, e.g., infected cells from a patient, can also include uninfected cells.

[0032] The term "diagnosis" is used herein to refer to the identification of a molecular or pathological state, disease or condition.

[0033] The term "prognosis" is used herein to refer to a prediction of the likelihood of disease progression (eg, progression of an infection), including recurrence, drug resistance, and the like.

[0034] The term "prediction" is used herein to refer to the act of foreseeing or estimating based on observation, experience, or scientific reasoning. In one example, a physician may predict the likelihood that a patient will survive.

[0035] The terms "specific binding," "specifically binds," and the like refer to non-covalent or covalent preferential binding to a molecule compared to other molecules or moieties in a solution or reaction mixture (e.g., an antibody specifically binds to a particular polypeptide or epitope compared to other available polypeptides / epitopes). In some embodiments, the affinity between a molecule and another molecule to which it specifically binds is greater than or equal to 10. -5 M or less (e.g., 10 -6 M or less, 10 -7 M or less, 10 -8 M or less, 10 -9 M or less, 10 -10 M or less, 10 -11 M and below, 10-12 M or less, 10 -13 M or less, 10 -14 M or less, 10 -15 M or less, or 10 -16 M or less) K D "Affinity" refers to the strength of binding, and increased binding affinity is characterized by a lower K D correlates with.

[0036] As used herein, the term "specific binding member" refers to a member of a specific binding pair (i.e., two molecules, usually two different molecules, where one of the molecules, e.g., the first specific binding member, specifically binds to the other molecule, e.g., the second specific binding member, via non-covalent means).

[0037] Anti-CD24 agents. The CD24 gene is located on human chromosome 6q21 and encodes two protein isoforms (CD24A and CD24B). CD24B (129 aa) has a longer, more distinct N-terminus than CD24A (83 aa). CD24A is the predominant isoform in HCC and plays a major role in cell proliferation, migration, and invasion. Commercially available antibodies, such as ML5 and SWA11, target a shared amino acid sequence and therefore do not distinguish between the CD24A and CD24B isoforms. However, in some embodiments, isoform-specific antibodies, such as CD24A or CD24B, can be selected.

[0038] CD24 is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein expressed at multiple stages of B cell development, beginning in the bone marrow pro-B cell compartment and continuing through mature surface Ig-positive B cells. Expression is very low or negative on plasma cells. It is also expressed in the majority of B-lineage acute lymphoblastic leukemias, B-cell CCLs, and B-cell non-Hodgkin's lymphomas. CD24 may play a role in regulating B-cell proliferation and maturation. Protein reference sequences include GenBank NP_001278666, NP_001278667, NP_001278668, NP_037362, and NP_001346013. Antibodies known to bind to human CD24 are known and commercially available, including, but not limited to, SWA11 (Creative Biolab), MA5-11833, 12-0247-42, anti-CD24 clone ML5 (Biolegend), SN3 A5-2H10 (also referred to as SN3), ALB 9, EPR19925, EPR3006(N), SWA21, SWA22, OKB2, and the like. Anti-CD24 agents may include, for example, antibodies that bind to human CD24, such as SWA11 or ML5. Anti-CD24 antibodies may bind, for example, to the LAP (leucine-alanine-proline) motif, as known in the art. Alternatively, antibodies specific for human CD24 may be generated. Humanized anti-CD24 antibodies are known in the art and are described, for example, in Weber et al., Clinical Exp. Immunol, 1993; Shapira et al. in: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016; 76(14 Suppl): Abstract nr 3805 and Sun et al. (2017) Oncotarget Vol. 8, (No. 31), pp: 51238-51252, each of which is specifically incorporated herein by reference. Humanized anti-CD24 antibodies include, for example, the humanized SWA11 antibody (as described in Arber Patent US8,614,301B2) and the humanized anti-CD24 antibody as described in CN103819561A.

[0039] CD24 is a heavily glycosylated surface glycophosphatidylinositol (GPI)-anchored protein, and as it lacks an intracellular signaling domain, CD24 mediates signaling pathways indirectly through interactions with cir- or trans-cell surface receptors. CD24 may play a crucial role in the differentiation and stress response of different cell types. Signal transduction can be triggered by the binding of lectin-like ligands to CD24 carbohydrates and can be transduced by the release of second messengers derived from GPI anchors or related molecules. In association with Siglec10, CD24 may be involved in the selective suppression of immune responses to danger-associated molecular patterns (DAMPs), such as HMGB1, HSP70, and HSP90. CD24 also binds to P-selectin.

[0040] As used herein, the term "anti-CD24 agent" refers to any agent that binds to CD24 on the surface of virus-infected cells and blocks or inactivates it, or impairs its function. In some embodiments, a suitable anti-CD24 agent (e.g., an anti-CD24 antibody, peptide, etc.) specifically binds to CD24 and reduces the interaction of CD24 with its ligand. In other embodiments, an anti-CD24 agent binds to and blocks the interaction of CD24 with P-selectin. In other embodiments, an anti-CD24 agent binds to and blocks the interaction of CD24 and Siglec10. In some cases, the anti-CD24 agent is an antibody, and in some cases, a humanized antibody. Small molecule compounds, including peptides, DNA, or RNA aptamers, that inhibit the binding of CD24 to one of its ligands are also considered to be anti-CD24 agents.

[0041] The effectiveness of a suitable anti-CD24 agent can be evaluated by assaying the agent. In a non-limiting example of such an assay, target cells are incubated in the presence or absence of a candidate agent, and target cell killing is measured. Agents (anti-CD24 agents) for use in the subject methods increase cell death by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to cell death in the absence of the candidate agent. Additionally, agents for use in the subject methods (anti-CD24 agents) reduce the level of HBV surface Ag (in the case of HBV) or other markers of viral infection (such as viral titer) by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to the absence of the candidate agent.

[0042] Anti-CD15 agents. CD15 is a carbohydrate adhesion molecule found on neutrophils that mediates phagocytosis and chemotaxis and is expressed in patients with Hodgkin's disease, some B-cell chronic lymphocytic leukemia, acute lymphoblastic leukemia, and most acute nonlymphocytic leukemias. This is the Lewis x and SSEA-1 (stage-specific embryonic antigen 1), it represents a marker for mouse pluripotent stem cells and plays an important role in cell adhesion and migration in the preimplantation embryo.

[0043] Anti-CD15 agents may include, for example, antibodies that bind to human CD15. Alternatively, antibodies specific to human CD15 may be generated. Antibodies are known in the art, for example, Roge et al. (2014) Appl Immuohistochem Mol Morphol. 2014 Jul; 22 (6): 449-58; Pellgrini et al. Haematologica. 2007 May; 92 (5): 708-9; Jegatheeswaran et al. J Immunol. 2019 Dec 1; 203 (11): 3037-3044, and are incorporated herein by reference.

[0044] As used herein, the term "anti-CD15 agent" refers to any agent that binds to CD15 on the surface of virus-infected cells and blocks or inactivates it, or impairs its function. In some embodiments, a suitable anti-CD15 agent (e.g., an anti-CD15 antibody, peptide, etc.) specifically binds to CD15 and reduces the interaction of CD15 with its ligand. In some cases, the anti-CD15 agent is an antibody, and in some cases, a humanized antibody. Small molecule compounds that inhibit the binding of CD15 to one of its ligands, including peptides, DNA, or RNA aptamers, are also considered to be anti-CD15 agents.

[0045] The effectiveness of a suitable anti-CD15 agent can be evaluated by assaying the agent. In a non-limiting example of such an assay, target cells are incubated in the presence or absence of a candidate agent, and target cell killing is measured. Agents (anti-CD15 agents) for use in the subject methods increase cell death by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to cell death in the absence of the candidate agent. Additionally, agents for use in the subject methods (anti-CD15 agents) reduce the level of HBV surface Ag (in the case of HBV) or other markers of viral infection (such as viral titer) by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to the absence of the candidate agent.

[0046] Anti-CD15 agents can specifically bind to and block the activity of CD15, for example, by blocking binding to one of its ligands (e.g., P-selectin, CD43, CD44, etc.) CD15 expression can be further reduced by agents that target its host synthetic enzymes, such as FUT3, FUT5, FUT6, FUT7, and / or the sialyltransferases ST3GAL3, ST3GAL4, and ST3GAL6.

[0047] Anti-CD104 Agent. CD104 (integrin beta-4 (ITB4)), encoded by the ITGB4 gene, is a receptor for laminin and plays a structural role in epithelial cells, and is required for keratinocyte polarity and motility. The cellular localization of the 1822-amino acid protein is predicted to be membrane-associated.

[0048] Anti-CD104 agents may include, for example, antibodies that bind to human CD104. Alternatively, antibodies specific for human CD104 can be generated. Antibodies are known in the art and are commercially available from several sources.

[0049] As used herein, the term "anti-CD104 agent" refers to any agent that binds to CD104 on the surface of virus-infected cells and blocks or inactivates it, or impairs its function. In some embodiments, a suitable anti-CD104 agent (e.g., an anti-CD104 antibody, peptide, etc.) specifically binds to CD104 and reduces the interaction of CD104 with its ligand. In some cases, the anti-CD104 agent is an antibody, and in some cases, a humanized antibody. Small molecule compounds that inhibit the binding of CD104 to one of its ligands, including peptides, DNA, or RNA aptamers, are also considered to be anti-CD104 agents.

[0050] The efficacy of a suitable anti-CD104 agent may be assessed by assaying the agent. In a non-limiting example of such an assay, target cells are incubated in the presence or absence of a candidate agent, and target cell killing is measured. Agents (anti-CD104 agents) for use in the subject methods increase cell death by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to cell death in the absence of the candidate agent. Additionally, agents (anti-CD104 agents) for use in the subject methods reduce the level of HBV surface Ag (in the case of HBV) or other markers of viral infection (such as viral titer) by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to the absence of the candidate agent.

[0051] Anti-CD133 agent. The CD133 antigen, also known as prominin-1, is a glycoprotein encoded in humans by the PROM1 gene. It is a member of a pentaspan transmembrane glycoprotein that specifically localizes to cell protrusions. When embedded in the cell membrane, the membrane topology of prominin-1 is such that the N-terminus extends into the extracellular space and the C-terminus resides in the intracellular compartment. The protein consists of five transmembrane segments, with the first and second and third and fourth segments connected by intracellular loops, and the second, third, and fourth and fifth transmembrane segments connected by extracellular loops.

[0052] The anti-CD133 agent may, for example, comprise an antibody that binds to human CD133. Alternatively, an antibody specific to human CD133 may be generated. Antibodies are known in the art (see, for example, Glumac et al. Prostate. 2018 Sep; 78(13): 981-991; Wang et al. Mol Pharm. 2019 Nov 4; 16(11): 4582-4593; Schmied et al. Cancers (Basel). 2019 Jun 7; 11(6): 789, etc.), each of which is incorporated herein by reference.

[0053] As used herein, the term "anti-CD133 agent" refers to any agent that binds to CD133 on the surface of virus-infected cells and blocks or inactivates it, or impairs its function. In some embodiments, a suitable anti-CD133 agent (e.g., an anti-CD133 antibody, peptide, etc.) specifically binds to CD133 and reduces the interaction of CD133 with its ligand. In some cases, the anti-CD133 agent is an antibody, and in some cases, a humanized antibody. Small molecule compounds that inhibit the binding of CD133 to one of its ligands, including peptides, DNA, or RNA aptamers, are also considered to be anti-CD133 agents.

[0054] The effectiveness of a suitable anti-CD133 agent can be assessed by assaying the agent. In a non-limiting example of such an assay, target cells are incubated in the presence or absence of a candidate agent, and target cell killing is measured. Agents (anti-CD133 agents) for use in the subject methods increase cell death by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to cell death in the absence of the candidate agent. In addition, agents (anti-CD133 agents) for use in the subject methods increase cell death by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to cell death in the absence of the candidate agent. The antibody reduces the level of HBV surface Ag (in the case of HBV) or other markers of viral infection (such as viral titer) by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to the presence of the antibody.

[0055] Anti-CD257 agent. The CD257 antigen, B-cell activating factor (BAFF), also known as tumor necrosis factor ligand superfamily member 13B, is a protein encoded by the TNFSF13B gene in humans. BAFF is also known as B-lymphocyte stimulator (BLyS), TNF- and APOL-related leukocyte-expressed ligand (TALL-1), and dendritic cell-derived TNF-like molecule (CD257 antigen; cluster of differentiation 257). This cytokine is a ligand for the receptors TNFRSF13B / TACI, TNFRSF17 / BCMA, and TNFRSF13C / BAFF-R. This cytokine is expressed in B-cell lineage cells and acts as a potent B-cell activator. It has also been shown to play an important role in B-cell proliferation and differentiation. BAFF is a 285-amino acid-long peptide glycoprotein that is glycosylated at residue 124. It is expressed as a membrane-bound type II transmembrane protein in various cell types, including monocytes, dendritic cells, and bone marrow stromal cells. The transmembrane form can be cleaved from the membrane, generating soluble protein fragments. BAFF is the natural ligand for three unusual tumor necrosis factor receptors: BAFF-R (BR3), TACI (transmembrane activator and calcium regulator and cyclophilin ligand interactor), and BCMA (B-cell maturation antigen), all of which have different binding affinities for it. These receptors are expressed primarily on mature B lymphocytes, and their expression changes depending on B-cell maturation (TACI is also found on a subset of T cells and BCMA on plasma cells). BAFF-R is involved in positive regulation during B-cell development. TACI binds to a BAFF-like protein called proliferation-inducing ligand (APRIL) with high affinity, making it the worst-case receptor. BCMA exhibits an intermediate binding phenotype and functions to varying degrees with either BAFF or APRIL.

[0056] Anti-CD257 agents may include, for example, antibodies that bind to human CD257. Alternatively, antibodies specific for human CD257 can be generated. Antibodies are known in the art and are commercially available from several sources.

[0057] As used herein, the term "anti-CD257 agent" refers to any agent that binds to CD257 on the surface of virus-infected cells and blocks or inactivates it, or impairs its function. In some embodiments, a suitable anti-CD257 agent (e.g., an anti-CD257 antibody, peptide, etc.) specifically binds to CD257 and reduces the interaction of CD257 with its ligand. In some cases, the anti-CD257 agent is an antibody, and in some cases, a humanized antibody. Small molecule compounds that inhibit the binding of CD257 to one of its ligands, including peptides, DNA, or RNA aptamers, are also considered to be anti-CD257 agents.

[0058] The effectiveness of a suitable anti-CD257 agent can be assessed by assaying the agent. In a non-limiting example of such an assay, target cells are incubated in the presence or absence of a candidate agent and target cell killing is measured. Agents (anti-CD257 agents) for use in the subject methods may reduce cell death by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, at least 220%, at least 240%, at least 260%, at least 280%, at least 290%, at least 300%, at least 310%, at least 320%, at least 330%, at least 340%, at least 350%, at least 360%, at least 370%, at least 380%, at least 390%, at least 400%, at least 410%, at least 420%, at least 430%, at least 440%, at least 450%, at least 460%, at least 470%, at least 480%, at least 490%, at least 500%, at least 510%, at least 520%, at least 530%, at least 540%, at least 550%, at least 560%, at least 570%, at least 580%, at least 590%, at least 600%, at least 610%, at least 610%, at least 620%, at least 630%, at least 640%, at least 650%, at least 660%, at least 670%, at least 680%, at least 690%, at least 700%, at least 71 In addition, agents for use in the subject methods (anti-CD257 agents) reduce the level of HBV surface Ag (in the case of HBV) or other markers of viral infection (such as viral titer) by at least 10% (e.g., at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 120%, at least 140%, at least 160%, at least 180%, at least 200%, or at least 300%) compared to the absence of the candidate agent.

[0059] The term "antibody" is used herein in the broadest sense and specifically encompasses monoclonal antibodies, polyclonal antibodies, monomers, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), heavy-chain-only antibodies, triple-chain antibodies, single-chain Fvs, single-domain antibodies, nanobodies, and the like, as well as antibody fragments, with or without pegylation, so long as they exhibit the desired biological activity (Miller et al. (2003) Jour. of Immunology 170:4854-4861). Antibodies may be murine, human, humanized, chimeric, or derived from other species. Antibodies, also called immunoglobulins, traditionally contain at least one heavy chain and one light chain; the amino-terminal domains of the heavy and light chains are variable in sequence and are therefore commonly referred to as variable region domains, or variable heavy (VH) or variable light (VL) domains. The two domains traditionally associate to form a specific binding region.

[0060] A "functional" or "biologically active" antibody or antigen-binding molecule is one that can exert one or more of its natural activities in structural, regulatory, biochemical, or biophysical events. For example, a functional antibody or other binding molecule may have the ability to specifically bind to an antigen, and the binding may then induce or alter cellular or molecular events such as signal transduction or phagocytosis. A functional antibody may also block ligand activation of a receptor, or act as an agonist or antagonist, or as an allosteric modulator.

[0061] The term antibody can refer to a full-length heavy chain, a full-length light chain, an intact immunoglobulin molecule, or an immunologically active portion of any of these polypeptides, i.e., a polypeptide comprising an antigen-binding site that immunospecifically binds to an antigen of a target of interest or a portion thereof, including, but not limited to, infected cells or cells that produce autoimmune antibodies associated with autoimmune disease. The immunoglobulins disclosed herein can contain any suitable Fc region, including, but not limited to, human or other mammalian, e.g., cynomolgus monkey, IgG, IgE, IgM, IgD, IgA, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2, or subclasses of immunoglobulin molecules, including hybrid Ig, hybrid Fc, and subclasses engineered with modified Fc portions that provide reduced or enhanced effector cell activity. The immunoglobulins can be derived from any species.

[0062] The term "variable" refers to the fact that certain portions of the variable domains vary widely in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called hypervariable regions in both the light and heavy chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs). Natural heavy and light chain variable domains each contain four FRs that primarily adopt a beta-sheet configuration, connected by three hypervariable regions that form loops connecting, and in some cases, part of, the beta-sheet structure. The hypervariable regions of each chain are held together in close proximity by the FRs, which, together with the hypervariable regions from the other chain, They contribute to the formation of the antigen-binding site of antibodies (see Kabat et al (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.). The constant domains are not directly involved in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity (ADCC).

[0063] The term "hypervariable region" as used herein refers to the amino acid residues of an antibody which are responsible for antigen binding. A hypervariable region may comprise amino acid residues from the "complementarity determining regions" or "CDRs" and / or those residues from the "hypervariable loops". "Framework Region" or "FR" residues are those variable domain residues other than the hypervariable region residues as herein defined.

[0064] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible natural mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method.

[0065] The term "antibodies" as used herein specifically includes "chimeric" antibodies in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical to or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Patent No. 4,816,567, and M. Orrison et al. (1984) Proc. Natl. Acad. Sci. USA, 81:6851-6855). Chimeric antibodies of interest herein include "primatized" antibodies containing variable domain antigen-binding sequences derived from a non-human primate (e.g., Old World monkey, ape, etc.) and human constant region sequences.

[0066] As used herein, an "intact antibody chain" includes a full-length variable region and a full-length constant region. An intact, "traditional" antibody includes an intact light chain and an intact heavy chain, as well as a light chain constant domain (CL) and heavy chain constant domains, CH1, hinge, CH2, and CH3, for secreted IgG. Other isotypes, such as IgM or IgA, may have different CH and CL domains. The constant domains may be native-sequence constant domains (e.g., human native-sequence constant domains) or amino acid sequence variants thereof. Intact antibodies may have one or more "effector functions," which refer to those biological activities attributable to the Fc constant region of an antibody (native-sequence Fc region or amino acid sequence variant Fc region). Examples of antibody effector functions include C1q binding, complement-dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADDC), phagocytosis, and downregulation of cell surface receptors. Constant region variants include those that alter the effector profile, such as Fc receptor binding.

[0067] Intact antibodies can be assigned to different "classes" depending on the amino acid sequence of the constant domain of their heavy chains. There are five major classes of intact immunoglobulin antibodies (IgA, IgD, IgE, IgG, and IgM), and some of these have "subclasses" (isotypes), e.g., IgG1, IgG2, IgG3, They can be further classified into IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different classes of antibodies are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. Ig forms include hinge-modified and hingeless forms (Roux et al. (1998) J. Immunol. 161: 4083-4090, Lund et al. (2000) Eur. J. Biochem. 267: 7246-7256, US 2005 / 0048572, US 2004 / 0229310). The light chains of antibodies from any vertebrate species can be assigned to one of two distinct types, called κ and λ, based on the amino acid sequence of their constant domains.

[0068] A "functional Fc region" has the "effector functions" of a native sequence Fc region. Exemplary effector functions include C1q binding, CDC, Fc receptor binding, ADCC, ADCP, down-regulation of cell surface receptors (e.g., B cell receptors), and the like. Such effector functions generally require the Fc region to interact with receptors, such as FcγRI; FcγRIIA; FcγRIIB1; FcγRIIB2; FcγRIIIA; FcγRIIIB receptors, and the recycling receptor FcRn, and can be assessed using various assays, e.g., as disclosed in the definitions herein. A "dead" or silenced Fc is one that has been mutated to retain activity, e.g., with respect to extended serum half-life, but does not bind to or activate low-affinity and high-affinity Fc receptors.

[0069] "Fv" is the minimum antibody fragment that contains a complete antigen-recognition and antigen-binding site. Fab fragments contain the constant domain of the light chain (CL) and the first constant domain of the heavy chain (CH1). Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region.

[0070] As used herein, an "antibody fragment," and all grammatical variations thereof, is defined as a portion of an intact antibody that contains the antigen-binding site or variable region of the intact antibody, which portion does not include the constant heavy chain domains of the Fc region of the intact antibody (i.e., CH2, CH3, and CH4, depending on the antibody isotype). Examples of antibody fragments include Fab, Fab', Fab'-SH, F(ab')2, and Fv fragments, diabodies; (1) any antibody fragment that is a polypeptide having a primary structure consisting of one uninterrupted sequence of consecutive amino acid residues (referred to herein as a "single-chain antibody fragment" or "single-chain polypeptide"), including, but not limited to, single-chain Fv (scFv) molecules; nanobodies or domain antibodies containing a single Ig domain from human or non-human species, or other specific single-domain binding modules, including, but not limited to, non-antibody binding proteins such as adnectins and anticalins, and multispecific or multivalent structures formed from antibody fragments.

[0071] As used in this disclosure, the term "epitope" refers to any antigenic determinant on an antigen to which the paratope of an antibody binds. 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.

[0072] As used herein, the term "label" refers to a detectable compound or composition that is directly or indirectly conjugated to a binding protein. The label may be detectable itself (a directly detectable label) (e.g., a radioisotope label or a fluorescent label), or the label may be indirectly detectable (e.g., in the case of an enzyme label, the enzyme may catalyze a chemical change in a substrate compound or composition and the reaction product is detectable).

[0073] As used herein, terms such as "correlate" or "correlated with" refer to a statistical association between instances of two events, where the events include numerical values, data sets, etc. For example, when the events include numerical values, a positive correlation (also referred to herein as a "direct correlation") means that as one increases, the other also increases. A negative correlation (also referred to herein as an "inverse correlation") means that as one increases, the other decreases.

[0074] Hepatitis B virus (HBV) is a hepatotropic virus that can cause severe liver diseases, including acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). Globally, approximately 250 million people suffer from chronic HBV infection, resulting in approximately 1 million deaths annually. HBV is an enveloped virus with a circular and partially double-stranded DNA genome of approximately 3.2 kb. After infection of hepatocytes, the HBV genome is delivered to the nucleus, where it is repaired to form covalently closed circular DNA (cccDNA), which then serves as a template to direct viral RNA transcription. This cccDNA is highly stable within the nucleus of infected hepatocytes, making chronic HBV infection difficult to treat, as cessation of treatment often leads to viral reemergence. Additionally, the HBV genome integrates into the host cell genome, and no specific treatment exists to reverse this process. A significant proportion of the vast amount of immunosuppressive HBsAg secreted by infected cells is derived from the integrated genome.

[0075] The viral genome is compact and encodes four overlapping genes, designated S, C, P, and X. The S gene encodes the viral envelope protein known as the surface antigen (HBsAg). The C gene encodes the 21-kDa core protein and the 25-kDa precore protein. The core protein packages its own mRNA, also known as pregenomic RNA (pgRNA), to form a core particle displaying the core antigenic determinant (i.e., core antigen, HBcAg). The precore protein contains the entire core protein sequence plus a 29-amino acid amino-terminal extension. The secreted precore protein derivative is known as the e antigen (HBeAg). Spontaneous loss of HBcAg and development of antibodies against HBeAg (known as HBcAg seroconversion) have been observed in many chronic HBV patients and is often preceded by a flare of hepatitis due to an increased cytotoxic T lymphocyte (CTL) response. The P gene encodes the viral DNA polymerase, which is also a reverse transcriptase, while the X gene encodes a regulatory protein with multiple functions, including enhancing viral gene expression and replication. After the formation of the viral core particle, the HBV pgRNA is reverse transcribed by the viral DNA polymerase, which is then packaged into a circular and partially double-stranded DNA genome. The core particle then interacts with HBsAg within the intracellular membrane to form mature viral particles, which are then released from infected hepatocytes. HBsAg can often be released from cells as empty subviral particles during massive virion overgrowth.

[0076] HBV causes chronic infection in approximately 250 million people worldwide. Recent research has generated a wealth of information to understand how HBV evades host immunity and establishes persistence. HBV can utilize type I IFN responses and suppress NK cells to enhance its replication. It can also exploit the infant's developed immune system and gut microbiota to educate fetal immunity to promote persistence in patients after vertical transmission. Studies by various groups have pointed to the important role of Kupffer cells in promoting HBV clearance and persistence. Current treatments for HBV infection include IFN-α or its pegylated derivatives and nucleoside / nucleotide analogs. These drugs are unsatisfactory because they produce durable responses in only a small proportion of patients.

[0077] Hepatitis D (delta hepatitis) is a small, spherical, enveloped virus. Hepatitis B virus (HDV) is a disease caused by hepatitis B virus (HBV). HDV can only be transmitted in the presence of hepatitis B virus (HBV) and is therefore considered a subviral satellite. HDV infection can occur through coinfection with HBV or in the context of chronic hepatitis B or hepatitis B carrier states (superinfection). Both superinfection and HDV coinfection result in more severe complications compared with HBV infection alone. These complications include a higher likelihood of liver failure and rapid progression to cirrhosis in acute infection and an increased risk of developing liver cancer in chronic infection.

[0078] HDV is a small, spherical virus with a diameter of 36 nm. It possesses an outer envelope containing three HBV envelope proteins (large, middle, and small hepatitis B surface antigens) and host lipids surrounding an internal nucleocapsid. The nucleocapsid contains a 1,679-nucleotide single-stranded circular RNA and approximately 200 molecules of hepatitis D antigen (HDAg) per genome. The central region of the HDAg has been shown to bind to RNA. The HDV genome exists as an enveloped, negative-sense, single-stranded, closed circular RNA. Its nucleotide sequence is 70% self-complementary, allowing the genome to form a partially double-stranded, rod-like RNA structure.

[0079] HDV is known to produce one protein, HDAg, which exists in two forms: a 27 kDa large HDAg and a 24 kDa small HDAg. The N-terminus of the two forms is identical, but the C-terminus of the large HDAg differs by an additional 19 amino acids. Both isoforms are normally produced from the same reading frame, containing a UAG stop codon at codon 196, which produces only the small HDAg (HDAg-S). However, editing by the cellular enzyme adenosine deaminase-1 changes the stop codon to UGG, allowing the production of the large HDAg (HDAg-L). HDAg-S is produced early in infection, enters the nucleus, and supports viral replication. HDAg-L is produced later in infection, functions as an inhibitor of viral replication, and, in its prenylated form, is required for viral particle assembly. RNA editing is important for the viral life cycle because it regulates the balance between viral replication and virion assembly.

[0080] Hepatitis D is transmitted in a similar manner to hepatitis B. Transmission is primarily restricted to people at high risk of hepatitis B infection, particularly injection drug users and those receiving clotting factor concentrates, but is also sexually transmitted. More than 15 million people are co-infected worldwide. HDV is rare in most developed countries and is mostly associated with intravenous drug use. However, HDV is much more common in the Mediterranean region, sub-Saharan Africa, the Middle East, and northern South America.

[0081] Current established treatments for chronic hepatitis D include conventional or pegylated interferon alpha therapy. Pegylated interferon alpha can be effective in reducing viral load and disease severity while the drug is being administered, but the benefits generally cease when the drug is discontinued. The efficacy of this treatment typically does not exceed approximately 20%, and late relapses after treatment have been reported. Prenylation inhibitors such as lonafarnib and pegylated interferon lambda have also been shown to be effective against HDV in clinical trials.

[0082] Hepatitis C virus (HCV) is a small, enveloped, single-stranded, positive-sense RNA virus. It belongs to the genus Hepacivirus in the family Flaviviridae. There are seven major genotypes of HCV, known as genotypes 1 through 7. The genotypes are divided into several subtypes, with the number of subtypes depending on the genotype. In the United States, approximately 70% of cases are caused by genotype 1 and 20% by genotype 2. Approximately 1% are caused by each of the other genotypes, and genotype 1 is most common in South America and Europe.

[0083] Hepatitis C is an infectious disease caused by the hepatitis C virus (HCV), which primarily affects the liver. Initial infection often presents with mild or no symptoms. Occasionally, symptoms include fever, dark urine, abdominal pain, and yellowish skin. The virus remains in the liver in approximately 75% to 85% of initially infected individuals. The early stages of chronic infection usually present no symptoms. However, over the years, it often leads to liver disease, sometimes cirrhosis. In some cases, patients with cirrhosis develop serious complications, such as liver failure, liver cancer, or dilated blood vessels in the esophagus and stomach.

[0084] HCV is spread primarily through intravenous drug use, poorly sterilized medical equipment, needlestick injuries in health care, and blood-to-blood contact associated with blood transfusions. Using blood tests, the risk from blood transfusions is less than 1 in 2 million. It can also be transmitted from an infected mother to her baby at birth. It is not spread by superficial contact. Diagnosis is by blood tests that look for either the virus or antibodies to its RNA. Testing is recommended for all at-risk individuals.

[0085] There is no vaccine for hepatitis C. Prevention includes harm reduction efforts among people who use drugs intravenously and screening of blood donations. Chronic infection can be cured approximately 95% of the time with antiviral medications such as sofosbuvir or simeprevir. Pegylated interferon and ribavirin were early developmental treatments with cure rates of less than 50% and greater side effects. People who develop cirrhosis or liver cancer may require a liver transplant; hepatitis C is the leading reason for liver transplantation, but the virus usually recurs after transplant.

[0086] As of 2015, an estimated 143 million people (2%) worldwide were infected with hepatitis C. Approximately 11 million new cases occurred in 2013. It occurs most commonly in Africa, Central Asia, and East Asia. Hepatitis C resulted in approximately 167,000 deaths from liver cancer and 326,000 deaths from cirrhosis in 2015.

[0087] Human papillomaviruses (HPVs) are small, double-stranded, circular DNA viruses with genomes of approximately 8,000 base pairs. The HPV life cycle strictly follows the differentiation program of host keratinocytes. HPV virions infect epithelial tissues via micro-abrasions, whereby they associate with putative receptors such as alpha integrins, laminins, and annexin A2, leading to virion entry into basal epithelial cells via clathrin-mediated endocytosis and / or caveolin-mediated endocytosis, depending on the HPV type. At this point, the viral genome is transported to the nucleus by an unknown mechanism, establishing a copy number of 10–200 viral genomes per cell. A sophisticated transcriptional cascade then ensues as host keratinocytes begin to divide and become increasingly differentiated in the upper layers of the epithelium. The two major oncoproteins of high-risk HPV types are E6 and E7. The HPV genome consists of six early (E1, E2, E4, E5, E6, and E7) open reading frames (ORFs), two late (L1 and L2) ORFs, and a non-coding length control region (LCR). After infection of host cells, the viral early promoter is activated, transcribing a polycistronic primary RNA containing all six early ORFs. This polycistronic RNA then undergoes active RNA splicing to generate multiple isoforms of mRNA. One of the spliced ​​isoform RNAs, E6*I, functions as the E7 mRNA, which translates the E7 protein. However, viral early transcription is regulated by viral E2, and high E2 levels suppress transcription. The HPV genome is integrated into the host genome by disruption of the E2 ORF, resulting in the E7 gene. The viral genome prevents E2 repression by E6 and E7. Thus, integration of the viral genome into the host DNA increases expression of E6 and E7, promoting cell proliferation and the potential for malignancy. The level of E6 and E7 expression correlates with the type of cervical lesion that may ultimately develop.

[0088] More than 40 types are transmitted through sexual contact and infect the anal and genital tract. Risk factors for persistent sexually transmitted infection include early age at first sexual intercourse, multiple partners, smoking, and poor immune function. These types are typically spread by persistent skin-to-skin contact, with vaginal and anal intercourse being the most common methods. Occasionally, HPV can be transmitted from mother to baby during pregnancy. HPV is thought to cause cancer both by integrating into DNA and by non-integrating episomes.

[0089] Most HPV infections cause no symptoms and resolve spontaneously. In some individuals, HPV infection persists, resulting in warts or precancerous lesions. Precancerous lesions increase the risk of cervical, vulvar, vaginal, penile, anal, oral, or throat cancer. Approximately 12 HPV types (including types 16, 18, 31, and 45) are referred to as "high-risk" types because persistent infection has been linked to cancers such as oropharyngeal, laryngeal, vulvar, vaginal, cervical, penile, and anal cancers. All of these cancers involve sexual transmission of HPV to stratified epithelial tissue. Individuals infected with both HPV and HIV are at increased risk of developing cervical or anal cancer. Nearly all cervical cancers are caused by two HPV types, HPV16 and HPV18, accounting for 70% of cases. 60% to 90% of the other cancers listed above are also HPV-related. HPV6 and HPV11 are common causes of genital warts and laryngeal papillomatosis.

[0090] HPV is the most common sexually transmitted disease worldwide. In 2018, an estimated 569,000 new cases and 311,000 deaths were attributed to cervical cancer worldwide, approximately 85% of which occurred in low- and middle-income countries. In the United States, approximately 30,700 cases of HPV-related cancer occur each year. Approximately 1% of sexually active adults have genital warts.

[0091] Herpes simplex virus (HSV) (human herpesvirus types 1 and 2) causes recurrent infections that typically affect the skin, mouth, lips, eyes, and genitals. Common severe infections include encephalitis, meningitis, neonatal herpes, and disseminated infections in immunocompromised patients. Mucocutaneous infections cause clusters of small, painful vesicles on an erythematous base. Diagnosis is clinical, with laboratory confirmation by culture, PCR, direct immunofluorescence, or serology. Treatment is symptomatic; antiviral therapy with acyclovir, valacyclovir, or famciclovir is helpful for severe infections and, if initiated early, for recurrent or primary infections.

[0092] Both types of herpes simplex virus (HSV), HSV-1 and HSV-2, can cause oral or genital infections. Most often, HSV-1 causes gingivostomatitis, cold sores, and keratitis. HSV-2 usually causes genital lesions.

[0093] HSV infection occurs through close contact with a person who is actively shedding the virus, which can occur from lesions or even when lesions are not apparent.

[0094] After initial infection, HSV remains dormant in nerve ganglia, from which it can periodically emerge and cause symptoms. Diseases caused by herpes simplex virus include mucocutaneous infections (most commonly), including genital herpes, eye infections (herpes keratitis), CNS infections, and neonatal herpes. In patients infected with HIV, herpes sensitivities are more common. Infection can be particularly severe. Progressive and persistent esophagitis, colitis, perianal ulcers, pneumonia, encephalitis, and meningitis can occur.

[0095] Human cytomegalovirus (HCMV, HHV-5) belongs to the Herpesviridae family, Betaherpesvirinae subfamily, and Cytomegalovirus class. The HCMV genome consists of approximately 230,000 bp of double-stranded DNA. The genome is enclosed by an icosahedral capsid (100–110 nm in diameter, 162 capsomeres). Between the capsid and the viral envelope is a protein layer known as the tegument. The viral envelope is derived from the cytoplasmic membrane. At least eight different viral glycoproteins are embedded in a lipid bilayer. Mature virus particles have a diameter of 150–200 nm. Like all herpesviruses, HCMV is sensitive to low pH, lipid-dissolving agents, and heat. HCMV has a half-life of approximately 60 minutes at 37°C and is relatively unstable at -20°C.

[0096] After adsorption of the virus to the target cell using viral glycoproteins, the viral envelope fuses with the cell membrane, and the capsid is released into the cell and transported to the nucleus, where the genome is released. Transcription of IE proteins then occurs in the cell nucleus using the host cell's RNA polymerase II. The tegument protein of the infectious virus particle functions as a transactivator of IE genes. IE proteins regulate the following stages of viral replication and are also involved in cellular regulation, including the expression and transport of HLA antigens (class I MHC proteins) to the proteasome. IE proteins (especially the phosphoprotein pp65) can be used as an early marker of viral infection in cell culture. The E protein contains the HCMV-encoded DNA polymerase that interacts with the viral nucleotide kinase, and their activity can be specifically inhibited with antiviral agents.

[0097] In immunocompetent individuals, most HCMV infections are asymptomatic or produce mild symptoms not characteristic of the disease. HCMV enters the body through mucosal contact or parenterally (via cell-containing blood components or stem cell / organ transplants) and can result in common infections involving organisms such as encephalitis, retinitis, hepatitis, nephritis, splenomegaly, and colitis. The virus can be transmitted to the fetus / child via the placenta or via cervical or vaginal secretions and breast milk (prenatal and postnatal transmission). Sexual transmission via cervical secretions, semen, or saliva is also possible.

[0098] Congenital infection is usually caused by primary maternal infection during pregnancy, with an intrauterine infection rate of 40–50%. Furthermore, mothers who are HCMV seropositive before pregnancy can be infected with additional HCMV strains (secondary maternal infection), with an infection rate of 1% of all newborns of seropositive mothers. Approximately 7–10% of HCMV-infected infants develop disease with clinical symptoms such as petechiae, jaundice, hepatosplenomegaly, chorioretinitis, and in some cases permanent neurological damage (e.g., mental retardation, hearing impairment, and even deafness, motor disabilities), and the outcome is fatal in approximately 10% of cases.

[0099] Key pathogenic mechanisms for the development of HCMV disease in tissue or organ transplant recipients are, on the one hand, a lack of immunity and / or immunosuppression, and, on the other hand, reactivation of latent virus in the case of preexisting HCMV infection in the recipient. HCMV reactivation can be triggered or enhanced by, among other things, bacterial infection, graft-versus-host disease (GVHD), or interactions with other viruses (e.g., HHV-6), or increased cytokine production up to a "cytokine storm" during treatment with antilymphocyte antibodies. Consequently, in addition to serological status and the type and intensity of immunosuppressive treatment, other risk factors for HCMV disease are concurrent bacterial and fungal infections, hepatitis after liver transplantation, or GVHD after allogeneic stem cell transplantation.

[0100] HIV-infected individuals are very frequently seropositive for HCMV and usually have advanced immunosuppression. Symptoms develop as a result of reactivation of latent virus during HCMV infection and rarely occur only as a result of primary infection. The occurrence of clinical HCMV symptoms correlates with the severity of immunosuppression, with patients with CD4+ T lymphocyte levels below 50-100 / μl being particularly affected.

[0101] Other herpesviruses of interest include, for example, EBV, VZV, and HHV-8, all of which can bind to the host genome and may be associated with the development of cancer, such as the development of Kaposi's sarcoma, varicella-zoster (VZV).

[0102] Human immunodeficiency virus (HIV) is a species of lentivirus (a subgroup of retroviruses) that causes HIV infection and, over time, acquired immune deficiency syndrome (AIDS). Lentiviruses are transmitted as single-stranded, positive-sense, enveloped RNA viruses. Upon entry into target cells, the viral RNA genome is reverse-transcribed into double-stranded DNA by reverse transcriptase, a virus-encoded enzyme transported along with the viral genome within the virus particle. The resulting viral DNA is then imported into the cell nucleus and integrated into cellular DNA by the virus-encoded enzyme integrase and host cofactors. Once integrated, the virus enters a latent state, allowing the virus and its host cell to indiscriminately evade detection by the immune system for extended periods of time. The HIV virus can remain dormant in the human body for up to 10 years after primary infection, during which time the virus does not cause symptoms. Alternatively, the integrated viral DNA may be transcribed and host cell resources may be used to produce new RNA genomes and viral proteins, which are packaged and released from the cell as new viral particles that initiate a new replication cycle.

[0103] HIV is characterized by two types: HIV-1 and HIV-2. HIV-1 was the first virus discovered and is designated both as lymphoblastic leukemia-associated virus (LAV) and human T-lymphotropic virus type 3 (HTLV-III). HIV-1 is more virulent and infectious than HIV-2 and causes the majority of HIV infections worldwide.

[0104] The RNA genome consists of at least seven structural landmarks (LTR, TAR, RRE, PE, SLIP, CRS, and INS) and nine genes (gag, pol, and env; tat, rev, nef, vif, vpr, vpu, and possibly a tenth gene, tev, which is a fusion of tat, env, and rev) that encode 19 proteins. Three of these genes, gag, pol, and env, contain the information necessary to make the structural proteins for new virus particles. For example, env encodes a protein called gp160, which is cleaved in two by cellular proteases to form gp120 and gp41. The remaining six genes, tat, rev, nef, vif, vpr, and vpu (or, in the case of HIV-2, vpx), are regulatory proteins that control HIV's ability to infect cells, produce new copies of the virus, or cause disease.

[0105] HIV-1 entry into macrophages and CD4+ T cells is mediated through interactions between the virion envelope glycoprotein (gp120) and CD4 molecules on the target cell membrane and with chemokine coreceptors. Macrophage-tropic (M-tropic), or non-syncytia-inducing, strains of HIV-1 use the β-chemokine receptor CCR5 for entry and are therefore able to replicate in both macrophages and CD4+ T cells. This CCR5 coreceptor is used by nearly all primary HIV-1 isolates, regardless of viral genetic subtype.

[0106] Management of HIV / AIDS typically involves the use of multiple antiretroviral drugs. In many parts of the world, HIV has become a chronic disease, and progression to AIDS is becoming increasingly rare. HIV latency and the activation of CD4+ T cells, dendritic cells, and macrophages are key factors in the management of HIV. The resulting viral reservoir in the gut is a major barrier to viral eradication.

[0107] As used herein, "cancer" includes any form of cancer, including, but not limited to, solid tumor cancers (e.g., lung, prostate, breast, bladder, colon, ovarian, pancreatic, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head and neck squamous cell carcinoma, melanoma, neuroendocrine, etc.) and liquid cancers (e.g., blood cancers), carcinomas, soft tissue tumors, sarcomas, teratomas, melanomas, leukemias, lymphomas, and brain cancers, including minimal residual disease, and including both primary and metastatic tumors. Any cancer is suitable for treatment by the subject methods and compositions.

[0108] Cancer types are malignant tumors derived from epithelial tissue. Examples of carcinomas include, but are not limited to, adenocarcinoma (cancer that originates in glandular (secretory) cells), for example, breast cancer, pancreatic cancer, lung cancer, prostate cancer, and colon cancer can be adenocarcinoma, adrenocortical carcinoma, hepatocellular carcinoma, renal cell carcinoma, ovarian cancer, intraepithelial carcinoma, ductal carcinoma, breast cancer, basal cell carcinoma, squamous cell carcinoma, transitional cell carcinoma, colon cancer, nasopharyngeal carcinoma, multilocular cystic renal cell carcinoma, oat cell carcinoma, large cell lung carcinoma, small cell lung carcinoma, non-small cell lung carcinoma, etc. Cancer can be found in the prostate, pancreas, colon, brain (usually as secondary metastasis), lung, breast, skin, etc.

[0109] Soft tissue tumors are a highly diverse and rare group of tumors derived from connective tissue. Examples of soft tissue tumors include, but are not limited to: alveolar soft part sarcoma; angiomatoid fibrohistiocytoma; chondromyo-oxidative fibroma; osteochondrosarcoma; extraskeletal myxoid chondrosarcoma; clear cell sarcoma; desmoplastic small round cell tumor; dermatofibrosarcoma protuberans; endometrial stromal tumor; Ewing's sarcoma; fibromatosis (fibroid); fibrosarcoma (infantile); gastrointestinal stromal tumor; giant cell tumor of bone; tenosynovitis giant cell tumor; inflammatory myofibroblastoma; uterine fibroids; leiomyosarcoma; lipoblastoma; typical lipoma; spindle cell or pleomorphic lipoma; atypical lipoma; chondroid lipoma; well-differentiated liposarcoma: myxoid / round cell liposarcoma; pleomorphic liposarcoma; myxoid malignant fibrohistiocytoma; high-grade malignant fibrohistiocytoma; myxofibrosarcoma; malignant peripheral nerve sheath tumor; mesothelioma; neuroblastoma; osteochondroma; osteosarcoma; primitive neuroectodermal tumor; alveolar rhabdomyosarcoma; embryonal rhabdomyosarcoma; benign or malignant nerve sheath tumor; synovial sarcoma; Evan's tumor; nodular fasciitis; fibroid fibromatosis: solitary fibrous tumor; dermatofibrosarcoma protuberans (DFSP); angiosarcoma; epithelioid hemangioendothelioma; giant cell tumor of tendon sheath (TGCT): pigmented villonodular synovitis (PVNS); fibrous dysplasia: myxofibrosarcoma; fibrosarcoma; synovial sarcoma; malignant peripheral nerve sheath tumor; neurofibroma; pleomorphic adenoma of soft tissue, tumors derived from fibroblasts, myofibroblasts, histocytes, vascular / endothelial cells and nerve sheath cells.

[0110] Sarcoma is a rare type of cancer that originates in bone or the soft tissues of the body, including mesenchymal cells, such as cartilage, fat, muscle, blood vessels, fibrous tissue, or other connective or supportive tissues. The different types of sarcoma are based on where the cancer forms. For example, osteosarcoma forms in bone, liposarcoma forms in fat, and rhabdomyosarcoma forms in muscle. Examples of sarcomas include, but are not limited to: Askin tumor; botryoid sarcoma; chondrosarcoma; Ewing sarcoma; malignant hemangioendothelioma; malignant schwannoma; osteosarcoma; and soft tissue sarcomas (e.g., alveolar soft part sarcoma; angiosarcoma; cystosarcoma; dermatofibrosarcoma protuberances (DFSP); desmoid tumor; desmoplastic small round cell tumor; epithelioid sarcoma; extraskeletal chondrosarcoma; extraskeletal osteosarcoma; fibrosarcoma; gastrointestinal stromal tumor (GIST); hemangiopericytoma; angiosarcoma; Kaposi's sarcoma; leiomyosarcoma; liposarcoma; lymphangiosarcoma; malignant peripheral nerve sheath tumor (MSTPN); neurofibrillary sarcoma; synovial sarcoma; malignant fibrous histiocytoma, and similar sarcomas).

[0111] Hematopoietic malignancies are leukemia, lymphoma, and myeloma. Leukemia is a cancer that begins in the blood-forming tissues, such as the bone marrow, and causes large numbers of abnormal blood cells to be produced and enter the bloodstream. Examples of leukemia include, but are not limited to: acute myeloid leukemia (AML), acute lymphoblastic leukemia (AML), and myeloma. allogeneic leukemia (ALL), chronic myeloid leukemia (CML), and chronic lymphocytic leukemia (CLL).

[0112] Lymphoma is a cancer that develops in cells of the immune system. For example, lymphoma can arise in bone marrow-derived cells that normally mature in the lymphatic system. There are two basic categories of lymphoma. One type, Hodgkin lymphoma (HL), is characterized by the presence of a type of cell called Reed-Sternberg cells. Currently, six types of HL are recognized. Examples of Hodgkin lymphoma include nodular sclerosis, classical Hodgkin lymphoma (CHL), mixed cellularity CHL, lymphocyte-depleted CHL, lymphocyte-rich CHL, and nodular lymphocyte-predominant HL. Another category of lymphoma is non-Hodgkin lymphoma (NHL), which includes a large, diverse group of cancers of immune system cells. Non-Hodgkin lymphoma can be further divided into cancers with an inert (slow-growing) course and cancers with an aggressive (rapid-growing) course. Currently, 61 types of HL are recognized. Examples of non-Hodgkin's lymphomas include, but are not limited to, AIDS-related lymphoma, anaplastic large cell lymphoma, angioimmunoblastic lymphoma, blastic NK-cell lymphoma, Burkitt's lymphoma, Burkitt-like lymphoma (small non-cleaved cell lymphoma), chronic lymphocytic leukemia / small lymphocytic lymphoma, cutaneous T-cell lymphoma, diffuse large B-cell lymphoma, enteropathic T-cell lymphoma, follicular lymphoma, hepatosplenic gamma delta T-cell lymphoma, T-cell leukemia, lymphoblastic lymphoma, mantle cell lymphoma, marginal zone lymphoma, nasal T-cell lymphoma, childhood lymphoma, peripheral T-cell lymphoma, primary central nervous system lymphoma, transformed lymphoma, therapy-related T-cell lymphoma, and Waldenstrom's macroglobulinemia.

[0113] Brain cancer includes any cancer of brain tissue, including, but not limited to, glioma (e.g., glioblastoma, astrocytoma, oligodendroglioma, epithelioma, etc.), meningioma, pituitary adenoma, vestibular schwannoma, primitive neuroectodermal tumor (medulloblastoma), etc.

[0114] The "pathology" of cancer includes all phenomena that compromise the patient's well-being, including, but not limited to, abnormal or uncontrolled cell proliferation, metastasis, interference with the normal function of neighboring cells, release of abnormal levels of cytokines or other secretory products, suppression or exacerbation of inflammatory or immune responses, neoplasia, premalignancy, malignancy, and invasion of surrounding or distant tissues or organs such as lymph nodes.

[0115] As used herein, the terms "cancer recurrence" and "tumor recurrence," and grammatical variations thereof, refer to the further growth of neoplastic or cancerous cells after a cancer diagnosis. In particular, recurrence can occur when further cancerous cell growth occurs in cancerous tissue. "Tumor spread" similarly occurs when tumor cells spread to local or distant tissues and organs, and thus tumor spread encompasses tumor metastasis. "Tumor invasion" occurs when tumor growth spreads locally and impairs the function of the involved tissue by compressing, destroying, or preventing normal organ function.

[0116] As used herein, the term "metastasis" refers to the growth of a cancerous tumor in an organ or body part that is not directly connected to the organ of the original cancerous tumor.Metastasis is understood to include micrometastasis, which is the presence of an undetectable amount of cancerous cells in an organ or body part that is not directly connected to the organ of the original cancerous tumor.Metastasis can also be defined as several steps in the process, such as the detachment of cancer cells from the original tumor site and the migration and / or invasion of cancer cells to other parts of the body.

[0117] [Composition] Pharmaceutical Compositions. Suitable anti-CD24 agents are pharmaceutical compositions suitable for therapeutic use, e.g., human treatment. Alternatively, or in combination, agents specific for CD15, CD47, CD104, CD133, or CD257 can be provided for therapeutic use. In some embodiments, pharmaceutical compositions of the present invention comprise one or more therapeutic entities of the present disclosure and include a pharmaceutically acceptable carrier, a pharmaceutically acceptable salt, a pharmaceutically acceptable excipient, and / or an ester or solvate thereof. In some embodiments, the use of an anti-CD24 agent involves combination with another therapeutic agent (e.g., another anti-infective agent). Therapeutic formulations containing anti-CD24 agents can be prepared by mixing the agent(s) having the desired purity with a physiologically acceptable carrier, a pharmaceutically acceptable salt, an excipient, and / or a stabilizer (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) (e.g., in the form of a lyophilized formulation or aqueous solution). Compositions containing anti-CD24 agents can be formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the schedule of administration, and other factors known to a physician.

[0118] "Pharmaceutically acceptable excipient" means an excipient that is generally safe, non-toxic, and useful in preparing the desired pharmaceutical composition, and includes excipients acceptable for veterinary and human pharmaceutical use. Such excipients can be solid, liquid, semi-solid, or, in the case of an aerosol composition, gaseous.

[0119] Solid formulations for oral administration may be in any form that allows antibody molecules to be accessible to patients in prescribed amounts and for prescribed periods of time. Oral formulations can take a number of solid formulation forms, including, but not limited to, tablets, capsules, or powders. Alternatively, solid formulations may be lyophilized and brought into solution prior to administration, either as a single dose or multiple doses. Antibody compositions should generally be formulated within a biologically relevant pH range and may be buffered to maintain an appropriate pH range during storage. Both liquid and solid formulations generally require storage at lower temperatures (e.g., 2-8°C) to maintain stability for longer periods.

[0120] Formulated antibody compositions, particularly liquid formulations, may contain bacteriostatic agents to prevent or minimize proteolysis during storage, including, but not limited to, effective concentrations (e.g., about 1% w / v) of benzyl alcohol, phenol, m-cresol, chlorobutanol, methylparaben, and / or propylparaben. Bacteriostatic agents may be contraindicated for some patients. Thus, lyophilized formulations can be reconstituted in solutions with or without such agents. Additional components may be added to buffers or solid antibody formulations, including sugars (including, but not limited to, polyhydroxy hydrocarbons such as sorbitol, mannitol, and glycerol and / or disaccharides such as sucrose, lactose, maltose, or trehalose) as cryoprotectants, and, in some cases, associated salts (including, but not limited to, NaCl, KCl, or LiCl). Such antibody formulations, particularly liquid formulations intended for long-term storage, rely on a range of osmolality, both of which are useful for promoting long-term stability.

[0121] The antibody molecule may be administered to an individual by any art-recognized route of administration, including, but not limited to, oral administration, administration by injection (including, in certain embodiments, intravenous, subcutaneous, intraperitoneal, or intramuscular injection), administration by inhalation, intranasal, or topical administration, either alone or in combination with other agents designed to aid in the treatment of the individual.

[0122] The route of administration may be determined by one skilled in the art, including, but not limited to, the desired physicochemical characteristics of the treatment. The temperature should be determined based on several considerations, as will be understood by those skilled in the art. Treatment may be provided, for example, at 2-8°C or higher, while still rendering the formulation useful for parenteral injection. Preservatives, stabilizers, buffers, antioxidants, and / or other additives may be included as needed. Formulations may include divalent cations (including but not limited to MgCl, CaCl, and MnCl) and / or non-ionic surfactants (including but not limited to polysorbate-80 (TWEEN 80™), polysorbate-60 (TWEEN 60™), polysorbate-40 (TWEEN 40™), and polysorbate-20 (TWEEN 20™)), polyoxyethylene alkyl ethers (Bri58™, Bri35™, and others such as TRITONX-100™, TRITONX-114™, NP40™, Span85, and the Pluronic® series of non-ionic surfactants (e.g., PLURONIC® 121). Any combination of such components forms a specific embodiment of the present disclosure.

[0123] "Pharmaceutically acceptable salts and esters" refers to salts and esters that are pharmaceutically acceptable and have the desired pharmacological properties. Such salts include salts that can be formed when acidic protons present in the compound are capable of reacting with inorganic or organic bases. Suitable inorganic salts include those formed with alkali metals, e.g., sodium and potassium, magnesium, calcium, and aluminum. Suitable organic salts include those formed with organic bases such as amine bases, e.g., ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. Also included are acid addition salts formed with inorganic acids (e.g., hydrochloric acid and hydrobromic acid) and organic acids (e.g., acetic acid, citric acid, maleic acid, and alkane- and arenesulfonic acids, such as methanesulfonic acid and benzenesulfonic acid). Pharmaceutically acceptable esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the compound, e.g., C 1-6 Examples include alkyl esters. When two acidic groups are present, the pharmaceutically acceptable salt or ester can be a mono-acid mono-salt or ester, or a di-salt or ester; similarly, when two or more acidic groups are present, some or all of the groups can be chlorided or esterified. The compounds named in this invention can exist in non-chlorinated or non-esterified form, or in chloride and / or esterified form, and the naming of the compounds is intended to include both the original (non-chlorinated and non-esterified) compound and its pharmaceutically acceptable salts and esters. Also, certain compounds named in this invention can exist in two or more stereoisomeric forms, and the naming of the compounds is intended to include all single stereoisomers and all mixtures of the stereoisomers (whether racemic or not).

[0124] The terms "pharmaceutically acceptable," "physiologically acceptable," and grammatical variations thereof are used interchangeably to refer to compositions, carriers, diluents, and reagents, and indicate that these materials can be administered to a human without producing undesirable physiological effects that would interfere with administration of the composition.

[0125] "Dosage unit" refers to a physically discrete unit suitable as a unitary dosage for a particular individual to be treated. Each unit can contain a predetermined quantity of active compound(s) calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit forms can be determined by (a) the inherent characteristics of the active compound(s) and the particular therapeutic effect(s) to be achieved, and (b) the limitations inherent in the technology for compounding the active compound(s).

[0126] [method] Exposing the individual to, in some cases, a therapeutically effective dose of an anti-CD24 agent for a period of time. provides methods for reducing viral load or other markers of chronic viral infection (e.g., HBV surface antigen in the case of HBV infection), including, but not limited to, elimination of virally infected cells in an individual. In some embodiments, the virus is a virus that causes chronic infection, for example, by integrating into the host genome. In some embodiments, the virus is a chronic hepatitis virus, e.g., hepatitis B, hepatitis C, or hepatitis D. In some embodiments, the virus is a lentivirus, e.g., HIV-1 or HIV-2. In some embodiments, the virus is a human papillomavirus. In some embodiments, the virus is a herpesvirus, e.g., herpes simplex virus (HSV), human cytomegalovirus (CMV), varicella-zoster virus (VZV), Kaposi's sarcoma-associated virus (HHV-8), Epstein-Barr virus (EBV), etc.

[0127] In other embodiments, methods are provided for reducing viral load or other markers of chronic viral infection (e.g., HBV surface antigen in the case of HBV infection), including, but not limited to, elimination of virally infected cells in an individual, in some cases, by contacting the individual with a therapeutically effective dose of an anti-CD15, anti-CD104, anti-CD133, anti-CD257 agent for a period of time sufficient to eliminate the virally infected cells.

[0128] An effective dose of an anti-CD24 agent can significantly reduce viral load or other markers of chronic viral infection, and in some cases, eradicate the presence of chronic viruses, including but not limited to HBV, at a dose that does not cause substantial toxicity to uninfected cells or the entire host. In some embodiments, treatment results in undetectable levels of viral antigens, e.g., HBsAg, in the individual's blood. The effect on uninfected cells, e.g., hepatocytes, can be monitored by the release of hepatocyte markers into the blood. Low levels of such markers are expected to be detected from the killing of infected cells, but at doses low enough to avoid dose-limiting toxicity. Adverse effects are assessed by conventional criteria, e.g., CDCAE. Using v5, it is desirable to have a level less than 3, but it can also be less than 2 or even less than 1.

[0129] The course of treatment can be less than about 12 weeks, less than about 8 weeks, less than about 4 weeks, e.g., 1-12 weeks, 2-12 weeks, 4-12 weeks, 4-8 weeks, etc. Administration can be weekly, twice weekly, every other day, daily, every two weeks, etc., and in some embodiments, weekly. In some embodiments, two or more courses of treatment are administered.

[0130] In other embodiments, the administration of an anti-CD24 agent is combined with the administration of an agent that specifically targets a second antigen upregulated on virus-infected cells. Markers of interest include, but are not limited to, CD15, CD104, CD257, CD105, CD133, and CD47. In some embodiments, the agent that binds to the second antigen is an antibody. In some embodiments, a bispecific antibody that targets CD24 and the second antigen is administered. In other embodiments, a bispecific antibody that binds CD24 with one arm and either CD3e (to redirect T cells), CD16a (to redirect NK cells), or other cell surface proteins on killer cells to target CD24-overexpressing populations may be used. In yet another embodiment, chimeric antigen receptor (CAR)-T cells, NKT cells, or NK cells that target CD24 with a CAR portion may be used to kill virus-containing cells.

[0131] An effective treatment reduces the number of virally infected cells in an individual, preferably eliminating the number of virally infected cells to undetectable levels. For example, the level of a viral antigen, such as HBsAg, is at least 1 log or more, 2 logs or more, and in some cases, less than 1 ng / ml, less than 0.5 ng / ml, less than 0.1 ng / ml, less than 0.05 ng / ml, or less than 1 ng / ml in the blood. HBsAg can be reduced by 1 log, 2 logs or more from baseline, as measured in IU / mL. IU / mL can be undetectable, for example, at less than 0.05, in some quantitative assays.

[0132] The terms "treatment," "treating," "treat," and the like are used herein generally to refer to obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in that a disease or its symptoms are completely or partially prevented, and / or therapeutic, in that the disease and / or adverse effects resulting from the disease are partially or completely stabilized or cured. The term "treatment" encompasses any treatment of disease in a mammal, particularly a human, and includes (a) treatment that prevents the disease and / or symptom(s) from occurring in a subject who may be predisposed to the disease or condition but has not yet been diagnosed as suffering from it, (b) treatment that inhibits the disease and / or symptom(s), i.e., prevents their development, or (c) treatment that alleviates the disease symptom(s), i.e., causes regression of the disease and / or symptom(s). Those in need of treatment include those already suffering from an infection (e.g., those with an infection, those with an infection (singular), those with an immune disorder, etc.), as well as those for which prevention is desired (e.g., those susceptible to an infection, those at high risk of an infection, those suspected of having an infection, etc.).

[0133] A therapeutic treatment is a treatment that is given to a subject prior to administration, and a prophylactic treatment is a treatment that is not given to a subject prior to administration. In some embodiments, the subject is likely to have or is suspected of having received the treatment prior to administration. In some embodiments, the subject is suspected of having received the treatment.

[0134] Examples of diseases that can be treated with anti-CD24 agents include, but are not limited to, viral infections, and may further include the development of cancers resulting from viral infections, such as liver cancer, oropharyngeal cancer resulting from HBV, HCV, HDV infection, and cancers of the reproductive system, such as cervical cancer, vulvar cancer, anal cancer, and penile cancer resulting from HPV infection, cancers associated with herpes virus infection, and the like.

[0135] As used herein, the term "infection" refers to any condition in which at least one cell of an organism (i.e., a subject) is infected by a virus. As used herein, the term "infectious agent" refers to a foreign biological entity. For example, infectious agents include, but are not limited to, bacteria, viruses, protozoa, and fungi. An infectious disease is a disease caused by an infectious agent. Some infectious agents do not cause discernible symptoms or disease under certain conditions, but may cause symptoms or disease under altered conditions. The subject method can be used to treat chronic viral infections, including, but not limited to, retroviruses, lentiviruses, hepadnaviruses, herpesviruses, poxviruses, human papillomaviruses, etc.

[0136] In some embodiments, the infectious disease is a chronic infection, i.e., an infection that is not eliminated by the host immune system within a period of up to one week, two weeks, etc. In some cases, a chronic infection involves the integration of pathogen genetic elements into the host genome, such as retroviruses, lentiviruses, hepatitis B viruses, etc. Infections treated by the methods of the present invention generally involve pathogens that have at least part of their life cycle within host cells, i.e., the intracellular phase. The methods of the present invention provide more effective elimination of infected cells.

[0137] The terms "co-administration," "administered simultaneously," and "combination" do not imply any specific time limit or other limitations. In addition, this includes administering two or more therapeutic agents (e.g., an anti-CD24 agent and an antiviral agent, and / or a target cell-specific antibody) in parallel, simultaneously, or sequentially. In one embodiment, the agents are present in a cell or subject's body at the same time or exert their biological or therapeutic effects simultaneously. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms. In certain embodiments, the first agent may be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, or following (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of the second therapeutic agent.

[0138] Administration of an anti-CD24 agent may be combined with the simultaneous administration of any number of additional antiviral agents. Such agents may include, for example, the entry inhibitor myrcludex-b, anti-NTBC antibodies, HBV nucleoside analogs (e.g., TDF, TAF, etc.), etc. Administration of an anti-CD24 agent may also be combined with a second antiviral agent, for example, an HBsAg release inhibitor (nucleic acid polymer), an HBV core inhibitor, an HBV (or HDV)-targeting siRNA, an immunomodulator (such as a TLR agonist), an RT or polymerase inhibitor, a therapeutic vaccine, etc. Other antiviral drugs can include interferons, including interferon alfa-2b, pegylated interferon alfa-2a, interferon lambda, entecavir, lamivudine, adefovir, telbivudine, tenofovir, sofosbuvir, ledipasvir, ombitasvir, paritaprevir, ritonavir, dasabuvir, grazoprevir, elbasvir, asunaprevir, decatasvir, or becrebuvir.

[0139] The anti-CD24 agent is optionally, but not necessarily, formulated with one or more agents that enhance activity or otherwise increase therapeutic effect. These are generally used in the same dosages and by the same routes of administration as used herein, or at 1-99% of the dosages previously used. In some embodiments, treatment is achieved by administering a combination (co-administration) of the subject's anti-CD24 agent and / or another agent that opsonizes target cells.

[0140] Treatment may also be combined with other active agents, such as antibiotics, cytokines, and antivirals. Antibiotic classes include penicillins, e.g., penicillin G, penicillin V, methicillin, oxacillin, carbenicillin, nafcillin, and ampicillin, in combination with β-lactamase inhibitors, cephalosporins, e.g., cefaclor, cefazolin, cefuroxime, and moxalactam, and carbapenems, monobactams, aminoglycosides, tetracyclines, macrolides, lincomycin, polymyxins, sulfonamides, quinolones, chloramphenicol, metronidazole, spectinomycin, trimethoprim, and vancomycin. Cytokines may also be used, such as interferon-γ, tumor necrosis factor-α, and interleukin-12. Antivirals, such as acyclovir and ganciclovir, may also be used in treatment.

[0141] A "therapeutically effective dose" or "therapeutic dose" is an amount sufficient to produce a desired clinical result (i.e., achieve therapeutic efficacy). A therapeutically effective dose can be administered in one or more administrations. In some embodiments, the anti-CD24 agent is an antibody specific for human CD24, which is optionally a chimeric or humanized monoclonal antibody. In some embodiments, the anti-CD24 antibody is administered at a dose of less than 8 mg / kg body weight, less than 2.5 mg / kg body weight, less than 1 mg / kg body weight, less than 0.75 mg / kg body weight, less than 0.5 mg / kg body weight, less than 0.25 mg / kg body weight, less than 0.1 mg / kg body weight, less than 0.05 mg / kg body weight, or less than 0.01 mg / kg body weight. A therapeutic dose can be, for example, 0.1-5 mg / kg, It can be 0.25 to 5 mg / kg, 0.5 to 5 mg / kg, 0.75 to 5 mg / kg, 1 to 5 mg / kg, or 0.1 to 2.5 mg / kg, 0.25 to 2.5 mg / kg, 0.5 to 2.5 mg / kg, 0.75 to 2.5 mg / kg, 0.1 to 1 mg / kg, 0.25 to 1 mg / kg, 0.5 to 1 mg / kg, 0.75 to 1 mg / kg, etc.

[0142] Dosage and frequency may vary depending on the half-life of the anti-CD24 agent. Those skilled in the art will appreciate that such guidelines will be adjusted for the molecular weight of the active agent, for example, in the use of antibody fragments, antibody conjugates, anti-CD24 agents, etc. Dosage may also vary for local administration, e.g., intranasal, inhalation, etc., or systemic administration, e.g., im, ip, iv, sc, etc.

[0143] The anti-CD24 agent may be administered by any suitable means, including topical, oral, parenteral, intrapulmonary, and intranasal. Parenteral infusion includes intramuscular, intravenous (bolus or slow drip), intraarterial, intraperitoneal, intrathecal, or subcutaneous administration. The anti-CD24 agent may be administered in any medically acceptable manner. This may include injection via intravenous, intravascular, intraarterial, subcutaneous, intramuscular, intratumoral, intraperitoneal, intracerebroventricular, intraepithelial, or other routes, as well as parenteral routes such as oral, nasal, ocular, rectal, or topical. Sustained-release administration is also specifically included in the present disclosure, by means such as depot injection or erodible implant. Local delivery is particularly contemplated, by means such as delivery via a catheter into one or more arteries, such as the renal artery or blood vessels supplying a local tumor.

[0144] As described above, anti-CD24 agents can be formulated with a pharmaceutically acceptable carrier (one or more natural or synthetic organic or inorganic components with which the subject agent is combined to facilitate its application). Suitable carriers include sterile saline, but other aqueous and non-aqueous isotonic sterile solutions and sterile suspensions known to be pharmaceutically acceptable are known to those skilled in the art. An "effective amount" refers to an amount that can ameliorate or slow the progression of a disease, degenerative, or damaged condition. An effective amount can be determined on an individual basis and is based, in part, on consideration of the symptoms to be treated and the results desired. An effective amount can be determined by one skilled in the art using such factors and only routine experimentation.

[0145] Anti-CD24 agents are often administered as pharmaceutical compositions containing an active therapeutic agent and another pharmaceutically acceptable excipient. The preferred form depends on the intended mode of administration and therapeutic use. The composition may also contain a pharmaceutically acceptable non-toxic carrier or diluent, defined as a vehicle commonly used to formulate pharmaceutical compositions for animal or human administration, depending on the desired formulation. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents include distilled water, physiological phosphate-buffered saline, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also contain other carriers, adjuvants, or non-toxic, non-therapeutic, non-immunogenic stabilizers.

[0146] The compositions can be prepared as injectables, either as liquid solutions or suspensions, or as solid forms suitable for solution or suspension in liquid vehicles prior to injection.The preparations can also be emulsified or encapsulated in liposomes or microparticles, such as polylactide, polyglycolide, or copolymers, for enhanced adjuvant effect, as described above (Langer, Science 249:1527, 1990 and Hanes, Advanced Drug Delivery Review 28:97-119-1997).The agents of the present invention can be administered in the form of depot injections or implant preparations, which can be formulated in a way that allows sustained or pulsatile release of the active ingredient.The pharmaceutical compositions are generally sterile, substantially isotonic, and comply with all U.S. Food and Drug Administration standards for drug and quasi-drug manufacturing practices. Formulated in full compliance with (GMP) regulations.

[0147] Toxicity of anti-CD24 agents can be assessed by standard pharmaceutical procedures in cell culture or experimental animals, e.g., LD 50 (lethal dose for 50% of the population) or LD 100 The lethal dose for 100% of the population can be determined by determining the lethal dose. The dose ratio between toxic and therapeutic effects is the therapeutic index. Data obtained from these cell culture assays and animal studies can be used to further optimize and / or define therapeutic and / or subtherapeutic dosage ranges (e.g., for use in humans). The exact formulation, route of administration, and dosage can be chosen by the individual physician in view of the patient's condition.

[0148] [kit] Kits for use in the subject methods are also provided. The subject kits can include an anti-CD24 agent. In some embodiments, the anti-CD24 agent is provided in a dosage form (e.g., a therapeutically effective dosage form). In the context of a kit, the anti-CD24 agent can be provided in liquid or solid form in any convenient package (e.g., a stick pack, a dose pack, etc.). The agents of the kit can be present in the same or separate containers. The agents may also be present in the same container. In addition to the above components, the subject kits can (in certain embodiments) further include instructions for practicing the subject methods. These instructions can be present in the subject kits in a variety of forms, one or more of which may be present in the kit. One form in which these instructions can be present is as printed information on a suitable medium or substrate, such as one or more sheets of paper on which the information is printed, in the kit packaging, in a package insert, etc. Yet another form in which these instructions can be present is as a computer-readable medium having the information recorded thereon, such as a diskette, a compact disc (CD), a flash drive, etc. Yet another form these instructions may take is a website address that can be used over the Internet to access information at the remote site.

[0149] While the present invention has been fully described, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the spirit or scope of the invention.

[0150] [experiment] The following examples are presented to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the following experiments are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.). However, some experimental error and deviation should be accounted for. Unless otherwise specified, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0151] The present invention has been described with reference to specific embodiments discovered or proposed by the inventors to comprise preferred modes for carrying out the invention. Those skilled in the art will understand, in light of this disclosure, that numerous modifications and variations can be made in the specific embodiments exemplified without departing from the intended scope of the invention. For example, codon redundancy allows for changes in the underlying DNA sequence without affecting the protein sequence. Biological functional equivalence considerations allow for changes in protein structure without affecting biological activity in kind or amount. All such modifications are intended to be within the scope of the appended claims.

[0152] [Example 1] To identify host cell targets for combating HBV infection, we investigated the expression of HBV-infected genes. We screened for host cell surface proteins that are upregulated by HBV. Using the Human Cell Marker Screening (PE) kit and Human Antibody Panel (PE) kit available from Biolegend, we compared surface protein expression levels between HBV-infected and non-infected cells. We used 2.2.15 cells (HepG2-derived cells harboring the HBV genome) as HBV-infected cells and compared the surface protein expression levels with non-infected HepG2 cells. Interestingly, we found that 2.2.15 cells have two distinct populations: one with high CD47 expression (CD47H) and one with low CD47 expression (CD47L). Using FACS, we isolated these two populations and compared the surface protein expression of both CD47H 2.2.15 and CD47L 2.2.15 cells with those of HepG2. We identified a set of surface markers that were elevated in both CD47H and CD47L 2.2.15 cells compared to HepG2 (Table 1). We also confirmed that both CD47L and C47H 2.2.1.5 cells contain comparable levels of HBsAg and DNA. As confirmation, we also transfected HepG2 cells with the HBV genome and a neomycin selection plasmid, selected clones carrying the HBV genome, and confirmed that these HBV-infected cells reliably upregulated the surface expression of these proteins.

[0153] [Table 1]

[0154] Among the surface proteins whose expression is most significantly increased in the presence of HBV is CD24. Using a qPCR assay, we found that CD24 is elevated at the transcriptional level. Our qPCR screening also revealed significantly elevated CD133 mRNA in HBV-infected cells compared with uninfected cells. We hypothesized that specific targeting of CD24 would be detrimental to HBV. One method for targeting CD24 is to use a monoclonal antibody specific for human CD24.

[0155] Other methods for targeting CD24 include high-affinity peptides fused to Fc, with or without pegylation, that bind to CD24, or that bind to human serum albumin (HSA), or that are conjugated to proteins that bind either IgG or HSA. Another potential method for targeting CD24 is to use one of the normal ligands, namely Siglec-10 or P-selectin, mutated (mutein-like) to act as antagonists fused to a half-life extension module such as Fc or HSA. CD24 expression can also be reduced with CRISPR, siRNA, or LNA targeting CD24. Small molecules that bind to CD24 are , can also be used to disrupt the CD24 axis, including DNA / RNA aptamers for CD24.

[0156] We prepared NSG mice with humanized livers using the RAAPID-TKG method. These mice contain chimeric livers harboring both mouse and human hepatocytes. The mice are deficient in B and T cells but retain macrophages. These mice can be efficiently infected with human hepatitis viruses, including HBV, HCV, and HDV. We infected these mice with HBV inocula from infected patients and established high levels of replication, as indicated by HBV DNA in serum, and HBsAg expression in serum.

[0157] Cohorts of mice were treated with anti-human CD24, anti-human CD47 (see Table 1), a combination of the two, or a vehicle control. The dose used was that commonly used to treat cancer with anti-human CD47, 200 μg / day administered IP (Weiskopf et al., Science (2013)). Mice tolerated the treatment well, and serum HBV DNA, HBsAg, and human albumin levels were monitored over time (see Table 2).

[0158] [Table 2]

[0159] In Table 2, UD means undetectable, N / A means unavailable, and PTW means 1 week after treatment. The indicated mice were treated daily with 200 μg of each indicated antibody. At the indicated time points, human albumin (hAlb, mg / ml), HBV DNA (copies / ml), and HBsAg (ng / ml) were measured. SWA11 and ML5 are examples of mouse monoclonal anti-human CD24 antibodies.

[0160] Surprisingly, anti-CD47 antibody alone, even when administered at a dose of 8 mg / kg (200 μg / mouse) daily for 4 weeks, had no effect on HBsAg or HBV DNA, likely reflecting a non-universal elevation of CD47 in HBV-infected cells. Anti-human CD24 was able to completely eradicate HBsAg. This was true when either SWA11 or ML5 anti-human was used. However, human albumin was also eradicated, resulting in a significant reduction in H This demonstrated that not only HBV-infected cells were killed, but also all human hepatocytes, including uninfected ones. This demonstrated that at this dose, anti-human CD24 was effective in eradicating HBV but also had undesirable toxicity to uninfected hepatocytes, resulting in the death of all human hepatocytes. Indeed, anti-human CD24 administered at 200 μg / mouse (8 mg / kg) resulted in a decrease in human albumin in uninfected mice, indicating undesirable toxicity to uninfected cells at this dose. We reasoned that the mice still appeared healthy because intact mouse hepatocytes remained in the chimeric liver, which were not targeted by anti-human CD24 and could support normal liver function.

[0161] This was confirmed by treating normal, immunocompetent mice (without any human hepatocytes) with the same dose of anti-mouse CD24 at 8 mg / kg, which resulted in high levels of mouse hepatocyte death, indicated by rapidly elevated ALT levels (see Table 3), and obvious clinical toxicity followed by death.

[0162] [Table 3]

[0163] BALB / c mice were treated for a set period with the indicated doses of rat anti-mouse CD24 and a combination of rat anti-mouse CD24 and rat anti-mouse CD47. Mouse behavior was scored, and ALT was measured at the end point. When BALB / c mice were treated daily for 4 days with 200 μg (8 mg / kg) of rat anti-mouse CD24 or a combination of rat anti-mouse CD24 and rat anti-mouse CD47, the mice became severely ill, and ALT levels were elevated in both treatment groups. Therefore, we found that anti-CD24 antibody at a dose of 8 mg / kg had unacceptable host toxicity.

[0164] However, we unexpectedly found that lower doses of anti-mouse CD24 were better tolerated in these standard mice, with ALT levels remaining normal at 25 μg (1 m / kg) twice weekly, indicating no death of uninfected hepatocytes (see Table 4). The low-dose regimen of anti-mouse CD24 is unexpectedly well tolerated in standard mice.

[0165] [Table 4]

[0166] Mice were treated twice weekly with a low-dose regimen (25 μg rat anti-mouse CD24 or 25 μg rat anti-mouse CD24 plus 100 μg rat anti-mouse CD47). After 2 weeks of treatment, all mice appeared healthy, and ALT levels in these mice were comparable to those in control mice, indicating a lack of hepatotoxicity.

[0167] When HBV-infected humanized mice were treated with a low-dose anti-human CD24 regimen, a progressive loss of HBsAg was still observed, but now without complete eradication of human albumin, indicating specific killing of infected hepatocytes with a relative lack of uninfected hepatocytes (see Table 5 ).

[0168] [Table 5-1]

[0169] [Table 5-2]

[0170] Chimeric mice with or without chronic HBV infection after inoculation with HBV-positive patient serum were treated with the indicated antibodies at different doses and frequencies (shaded cells indicate the treatment period). HBV infection status is labeled as "yes" (for chronic HBV infection) or "no" (for uninfected mice). "HDV" indicates that HBV-chronically infected chimeric mice were superinfected with HDV-positive patient serum. Human albumin (hAlb, mg / ml), HBsAg (ng / ml), and HDV RNA (copies / ml) in mouse serum were measured using the indicated methods. "UD" means undetectable.

[0171] HBV-uninfected chimeric mice (#562 and #563) maintained human albumin levels for two weeks. Similarly, HBV-uninfected chimeric mice (#567 and #571) were treated with mouse anti-human CD24 (25 μg or 1 mg / kg) or mouse anti-human CD24 plus anti-human CD47 (100 μg) for two weeks, but there was no significant decrease in human albumin levels. These data demonstrate that low-dose mouse anti-human CD24 (25 μg or 1 mg / kg) or mouse anti-human CD24 plus anti-human CD47 (100 μg or 4 mg / kg) is a safer dose for chimeric mice. Furthermore, a chimeric mouse chronically infected with HBV (#544) maintained HBsAg levels for two weeks without any antibody treatment. In contrast, when chimeric mice (#547 and #548) were treated twice weekly with mouse anti-human CD24 (100 μg or 4 mg / kg) or mouse anti-human CD24 plus anti-human CD47 (100 μg or 4 mg / kg), a significant decrease in human albumin was observed over a 2-week period. Therefore, subsequent chimeric mice were treated with reduced antibody doses and frequencies, e.g., 25 μg (1 mg / kg) (#562A and #543), 12.5 μg (0.5 mg / kg) (#550), and 6.25 μg (0.25 mg / kg) (#567A), every two weeks. At these doses, HBsAg was eradicated in chronically HBV-infected chimeric mice after only two weeks of treatment, and remained undetectable even after treatment. Furthermore, anti-human CD24 treatment eradicated HDV RNA and HBsAg in HDV-coinfected HBV chimeric mice (#563A).

[0172] These data demonstrate that a low-dose regimen can result in significant reductions of HBsAg and HBV DNA, which may be mediated in part through the killing of HBV-infected cells, in a more controlled and specific manner, making it ideal for treating HBV-infected patients. Because only a small proportion of a patient's total hepatocyte population is infected with HBV, and because uninfected hepatocytes can expand to replace HBV-infected hepatocytes that may be removed or killed by anti-human CD24, anti-human CD24 offers a novel and highly attractive breakthrough treatment for chronic HBV infection.

[0173] The desired benefits are: Rapid and controlled loss of HBsAg, which may help overcome HBsAg-mediated immunosuppression of the host anti-HBV immune response, further contributing to the control and eradication of HBV. Killing and elimination of HBV-infected hepatocytes, which highlights the most challenging aspect of HBV infection: the integrated HBV genome, which cannot be eliminated by current treatments. This could reduce the risk of HBV-related cancers. Very short treatment course. Only small doses of anti-human CD24 are required to achieve the desired treatment outcome. This is in stark contrast to current anti-HBV therapies, which must be taken lifelong because they only suppress HBV but do not eradicate HBsAg or HBV infection. Extremely convenient dosing. Anti-human CD24 can be administered once a week, or potentially less frequently, to achieve the desired anti-HBV effect. This contrasts with current therapies, which must be taken daily.

[0174] CD24 can also be elevated in cells infected with other viruses that cause chronic infections, such as hepatitis C virus (HCV), hepatitis delta virus (HDV), human immunodeficiency virus (HIV), cytomegalovirus (CMV), human papillomavirus (HPV), and herpes simplex virus (HSV), some of which are highly associated with subsequent cancer formation.

[0175] We tested the hypothesis that CD24 antibodies also have anti-cancer effects. We selected HBV-infected cells with high CD47 expression, which are known to form tumors in NSG mice within 6 weeks after subcutaneous flank injection. Prior to subcutaneous flank injection, we preincubated 1 million cells with 10 μg (20 μg / mouse or 0.8 mg / kg) of either anti-human CD24, anti-human CD15, anti-human CD47, a combination of anti-human CD24 and anti-human CD15, a combination of anti-human CD24, anti-human CD15, and anti-human CD47, or an IgG isotype control. Pretreatment with as little as 20 μg / mouse (0.8 mg / kg) of anti-human CD24 completely abolished tumor formation (Figure 1).

[0176] In addition to antiviral and anticancer effects, therapeutics targeting CD24 and / or its immune cell binding partner, Siglec-10, may also have clinical benefit in conditions where CD24 is inappropriately elevated and leads to effective clearance of pathological cells by immune cells, such as in fibrotic diseases and atherosclerosis, and where immune cells are ineffective at clearing disease cells.

[0177] NSG mice lack functional B, T, and NK cells but still possess functional macrophages. The antiviral effects observed in humanized mice may be partially mediated through phagocytic activity, as supported by in vitro phagocytosis assays performed using a mouse macrophage cell line (RAW264.7) and HepG2.2.1.5 cells (carrying the HBV genome). Here, administering anti-CD24 antibodies can enhance phagocytic clearance of HBV-infected cells.

[0178] We further determined whether antibodies targeting other surface markers elevated in HBV-infected cells could enhance phagocytic clearance. Indeed, antibodies targeting CD15, CD47, CD104, CD133, and CD257 increased phagocytosis of HepG2.2.1.5 cells, demonstrating that therapeutics targeting CD15, CD47, CD104, CD133, and CD257 can be effective antiviral drugs (Figure 2). Although anti-CD47 antibodies did not exhibit antiviral effects in our HBV-infected mice, this likely reflects the inconsistent elevation of CD47 on HBV-infected cells, as indicated by our 2.2.1.5 CD47H and CD47L populations. However, anti-CD15, anti-CD104, anti-CD133, and anti-CD257 agents are likely to have actual anti-HBV therapeutic effects, as these markers are elevated in both 2.2.1.5 CD47H and CD47L cells.

[0179] [material and method] Antibodies. Mouse anti-human CD24 (clone #ML5 and #SWA11) and rat anti-mouse CD24 were purchased from Biolegend (San Diego, CA) and Creative Biolab (Shirley, NY), respectively. Mouse anti-human CD24 (clone #B6H12) was purchased from BD Bioscience (San Jose, CA).

[0180] NOD.Cg-Prkdc scid II2rg tm1Wjl / SzJ background Bound-bearing, BALB / c, and NSG mice (catalog #005557) were purchased from Jackson Laboratory Inc. (Sacramento, CA, USA). All mice were housed in a pathogen-free barrier facility at Stanford University's Animal Research Facility, and irradiated mice were provided with food and autoclaved water ad libitum. All animal procedures were performed in accordance with protocols approved by the Stanford University Institutional Animal Care and Use Committee (IACUC).

[0181] Humanized mice using the RAAPID-TKG method. rAAV8-HSVtk (4 × 10 13 The rAAV8-HSVtk solution (vg / kg) was adjusted to 250 μl with saline and then inoculated into mice (approximately 25 g) via tail vein injection. Two to three weeks after rAAV8-HSVtk administration, mice were treated with two doses of ganciclovir (GCV) (catalog #315110, APP Pharmaceuticals, Schaumburg, IL) at 5 mg / kg (mouse body weight) at two-day intervals to induce injury. Elevated alanine aminotransferase (ALT) activity in the serum of treated mice, an indicator of liver injury, was measured using an Element DC Veterinary Chemistry Analyzer (catalog #6330-ALT, HESKA, Loveland, CO, USA). Mice with elevated ALT levels (>200 IU / L) were intrasplenic transplanted with cryopreserved human hepatocytes (lot #TVR) purchased from BioIVT (Baltimore, MD, USA). Four weeks after transplantation, human albumin in mouse serum was measured using a Human Albumin ELISA Quantitation Kit (Catalog #: E88-129, Bethyl Laboratories Inc., Montgomery, TX, USA) according to the manufacturer's protocol. Human hepatocyte engraftment efficiency was estimated in vivo by human albumin levels and, at the time of sacrifice, by calculating the area occupied by human hepatocytes in whole liver sections using Photoshop software 6.2 (Adobe, San Jose, CA, USA). Typically, six weeks after transplantation, chimeric mice were treated weekly with a hamster monoclonal anti-mouse CD95 antibody (200 μg / kg), an inducer of mouse hepatocyte-specific apoptosis, to maintain human albumin levels.

[0182] HBV and HDV infection of chimeric mice. Chimeric mice with >2 mg / ml human albumin in their serum were used for hepatitis virus infection. All hepatitis virus infections were performed by intravenous tail vein injection. To establish chronic HBV infection, chimeric mice were injected with 100 μL of human serum from an HBV-positive patient (lot #S1072). Two to three weeks after HBV inoculation, HBV DNA copy number was measured by qPCR as described below, and HBsAg was measured with an HBsAg quantification kit (Alpha Diagnostic Intl. Inc., San Antonio, TX). For hepatitis delta virus (HDV) coinfection, 10 per ml was used. 6 Chronically infected mice with >1U of HBV DNA were superinfected with 50 μL of HDV-positive patient serum. Three weeks after superinfection, mouse serum was collected, and HDV RNA in the mouse serum was measured by RT-PCR as described below. For antiviral studies, chronically HBV-infected chimeric mice were treated with various doses of mouse anti-human CD24 (clone #ML5 or #SWA11) at the indicated time points. HBV DNA and HBsAg, as well as HDV RNA in the case of superinfection, were monitored.

[0183] Virological Measurements. To determine HBV DNA copy numbers in mouse serum, HBV DNA from mouse serum samples (10 μl) was extracted using the Zymo Viral DNA Kit (Cat. #: D3017, Zymo Research, Irvine, CA, USA) according to the manufacturer's instructions. Concurrently, an AcroMetrix HBV (Cat. #: 965003, AcroMetrix, Benicia, CA, USA) standard control was serially diluted in 10-fold increments, extracted, and run in parallel with the HBV qPCR assay using a Bio-Rad CFX96 Real-Time System (Bio-Rad, Hercules, CA, USA). qPCR reaction conditions were performed as described using SssoAdvanced Universal Probes Supermix (Cat. #: 1725281, Bio-Rad, Hercules, CA, USA) and primers and probes synthesized by Integrated DNA Technologies (Coralville, IA, USA). HBV forward primers were used. Primer: 5′-AGTGTGGATTCGCACTCCT-3′, reverse primer: 5′-GAGTTCTTCTTCTAGGGGACCTG-3′, probe: 5′FAM-CCAAATGCCCCTATCTTATCAACACTTCC-IABFQ3′. HDV viremia was measured using a BioRad QX200 Digital Droplet Quantification was performed using a two-step PCR system (Bio-Rad Labs, Hercules, CA). HDV RNA from mouse serum (10 μl) and 10-fold serially diluted HDV-positive standard serum (10 μl) was isolated using the Zymo ZR Viral RNA Kit (Cat. #: R1035, Zymo Research, Irvine, CA, USA). Twenty microliters of eluted RNA was placed in a new PCR tube with 1.25 μL of RNAse-OUT (Cat. #: 10777-019, Thermofisher Scientific, Waltham, MA, USA), heated to 94°C for 2 minutes, and then immediately frozen on dry ice. This step was important for relaxing the RNA and achieving optimal reverse transcription. After thawing on wet ice, the RNA was purified using AccuPower RocketScript RT. The PCR mixture was combined with a premix (catalog number: K2104, Bioneer Corporation, Korea) and 50 fmol of the reverse primer (5′-GGTCGCATGGCATCTCCA-3′) under cycling conditions of 30°C for 5 min, 70°C for 2 h, and 95°C for 5 min. The ddPCR primer and probe sequences targeting highly conserved regions of the HDV genome were synthesized by Integrated DNA Technologies (Coralville, IA, USA) as follows: forward primer: 5′-GGCWCTCCCTTAGCCACCG-3′, reverse primer: 5′-GGTCGGCATGGCATCTCCA-3′, and probe: 5′FAM-CTCCTWCGGATGCCCAGGTCGGAC-IABFQ-3′.

[0184] Antibody treatment of chimeric mice chronically infected with HBV. Chimeric mice chronically infected with HBV were treated with the indicated IgG isotype control, mouse anti-human CD24, mouse anti-human CD47, or a combination of mouse anti-human CD24 and CD47. Control chimeric mice with or without HBV infection were included as controls. At the indicated time points, human albumin (hAlb, mg / ml), HBV DNA (copies / ml), and HBsAg (ng / ml), as well as HDV RNA (copies / ml) in cases of superinfection, were measured. Baseline data represent initial values ​​immediately prior to treatment. PTW data represent values ​​after that week of treatment. If HBsAg was still detectable in chimeric mice after 2 weeks of treatment, the chimeric mice were treated weekly with 25 μg of mouse anti-human CD24 for an additional 2 weeks.

[0185] [Example 2] The safety, tolerability, and pharmacokinetics (PK) of the anti-CD24 antibody will be evaluated in an open-label, multiple-dose study in healthy volunteers and HBV-infected patients.

[0186] Seven initial single-dose cohorts, each with six participants, will be enrolled through stepwise and dose-escalation studies. Dosages will be 0.05 mg / kg, 0.1 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, and 3 mg / kg administered intravenously or subcutaneously weekly or at other intervals, as determined by the pharmacokinetics observed in the single-dose escalation studies. All participants will have weekly blood draws to assess CBC, CMP, and coagulation function for up to four weeks after dosing.

[0187] Dose-limiting toxicity (DLT) was defined as the lowest dose at which 1 / 6 participants experienced a level 3 or higher adverse event (AE) related to the investigational drug occurring within 28 days of administration, as assessed by CTCAE v5.0.

[0188] The anti-CD24 antibody is then administered weekly for 4 weeks at the three highest dose groups with less than DLT. All participants will have weekly blood draws to assess CBC, CMP, and coagulation function for up to 4 weeks after the last dose. The maximum tolerated dose (MTD) will be defined as the highest dose at which 0 / 6 participants experience a grade 3 or higher AE.

[0189] This MTD is chosen to evaluate the efficacy of anti-CD24 antibodies in treating patients with HBV.

[0190] All recruited HBV patients met criteria for treatment with nucleoside analogs, such as tenofovir disoproxil fumarate (DF), tenofovir alafenamide, or entecavir. All patients received nucleoside analog therapy for at least 3 months and continued the nucleoside analog for the duration of anti-human CD24 antibody therapy. HBV patients received weekly anti-CD24 antibody therapy at the MTD, 1 / 3 MTD, and 1 / 9 MTD for 4 weeks. All participants had weekly blood draws to evaluate CBC, CMP, Coag, HBV DNA, and quantitative HBVsAg.

[0191] [Example 3] We describe monoclonal antibody-mediated phagocytic clearance of HBV cell lines. In vitro phagocytosis of HBV-infected cells is enhanced by blocking multiple cell surface molecules. Virus-infected cells were labeled and mixed with macrophages in the presence of control or blocking antibodies. HepG2.2.15 cells were labeled with mCherry (red) by transducing a lentiviral vector containing the mCherry gene under the PGK promoter. RAW 264.7 cells were cultured in IMDM medium with 10% FBS and activated with 50 nM phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich) and 10 ng / mL M-CSF. After 2 days of activation, Raw 264.7 cells were detached and labeled with 0.1 mM calcein AM (green) for 30 minutes at 37°C. Labeled HepG2.2.15 and Raw264.7 cells were mixed at a ratio of 1:10 and seeded onto 8-well chamber slides. Cells were then rapidly allowed to adhere by low-speed centrifugation for 5 minutes. The cells were then cultured in IMDM containing 50 mM PMA and 10 ng / ml pigment epithelium-derived factor (PEDF). The designated antibodies were added to each well to a final concentration of 10–20 μg / mL. After 24 hours of co-culture, the cells were washed three times with PBS and mounted in antifade medium containing DAPI. Images were captured using a KEYENCE BZ-X710 All-in-One Fluorescence Microscope (KEYENCE Corp. of USA, Itasca, IL, USA).

[0192] Figures 2A to 2F show the results using the following blocking or control antibodies: A. IgG2a negative control, B. anti-CD24, C. anti-CD15, D. anti-CD47, E. anti-CD104, F. anti-CD133, and G. anti-CD257.

[0193] Virus-infected HepG2.2.15 cells were labeled with mCherry (red). Macrophages, RAW264.7, were labeled with Calcein AM (green). Arrows indicate where in vitro phagocytosis of HepG2.2.1.5 cells by RAW264.7 cells occurred in the presence of different antibodies (anti-CD24, anti-CD15, anti-CD47, anti-CD104, anti-CD133, and anti-CD257) (detected by the presence of red dots (fragments of HepG2.2.1.5 cells) within green-labeled RAW264.7 cells). No phagocytosis was observed in the presence of an IgG2a control antibody.

[0194] These data suggest that, at least in vitro, these blocking antibodies play a role in increasing phagocytosis of virus-infected cells. Surprisingly, for anti-CD24 antibodies in particular, the antiviral-effective doses were not significantly different from the in vitro phagocytosis results. The antiviral dose was determined in an in vivo experiment as shown in Example 1.

[0195] [Cross reference] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 860,533, filed June 12, 2019, which applications are incorporated herein by reference in their entireties.

Claims

1. 1. Use of an anti-CD24 antibody in the manufacture of a medicament for use in a method for reducing hepatitis virus in an individual, comprising: the method comprises contacting the individual with a therapeutically effective dose of the anti-CD24 antibody for a period of time sufficient to reduce the amount of the hepatitis virus; The use of an anti-CD24 antibody, wherein the hepatitis virus is one or more of hepatitis B virus (HBV) and hepatitis delta virus (HDV).

2. the individual is a human and the anti-CD24 antibody specifically binds to human CD24, and / or The therapeutically effective dose is a dose sufficient to kill infected cells without unacceptable toxicity. Use of the anti-CD24 antibody of claim 1.

3. The therapeutic dose of the antibody is i) is less than 8 mg / kg administered over 24 hours; ii) is less than 8 mg / kg administered for 7 days; iii) is less than 1 mg / kg administered over 24 hours; or iv) is less than 1 mg / kg administered for 7 days; Use of the anti-CD24 antibody of claim 2.

4. i) the course of treatment is 1 to 24 weeks, 1 to 8 weeks, or 1 to 4 weeks; ii) the anti-CD24 antibody is administered in combination with an agent that specifically targets a second antigen upregulated on virally infected cells; Optionally, a) said second antigen is selected from CD15, CD104, CD257, CD105, CD133, and CD47; and / or b) the anti-CD24 antibody is a bispecific antibody that specifically binds to CD24 and the second antigen upregulated on virus-infected cells; iii) the anti-CD24 antibody is administered in combination with an agent that reduces reinfection of cells by the virus; Optionally, the agent that reduces reinfection of a cell by the virus is selected from the entry inhibitor myrcludex-b, and a nucleoside analog. Further optionally, the nucleoside analog is tenofovir disoproxil fumarate (DF), tenofovir alafenamide, or entecavir; and / or iv) the anti-CD24 antibody is administered in combination with a second antiviral agent; Optionally, the second antiviral agent is one or more of an HBsAg release inhibitor, an HBV core inhibitor, an siRNA targeting HBV or HDV, an immunomodulatory agent, a prenylation inhibitor, an RT or polymerase inhibitor, and a therapeutic vaccine. Use of an anti-CD24 antibody according to any one of claims 1 to 3.

5. The use of an anti-CD24 antibody according to any one of claims 1 to 4, wherein the virus causes a chronic infection.

6. i) the virus is a combination of HBV and HDV; ii) a) the HBV genome is cleared from the individual following treatment; b) HBsAg is at a level of less than 1 IU / mL or 1 ng / mL in the individual's blood (serum or plasma) after treatment; c) the concentration of HBsAg in the individual's blood is reduced by at least 1 log after treatment compared to the pre-treatment concentration; or d) the individual achieves HBsAg seroconversion to anti-HBs positive, and / or iii) virally-induced cancer is prevented or treated in said individual; Use of the anti-CD24 antibody according to claim 5.

7. the anti-CD24 antibody is a humanized anti-CD24 antibody, and / or The anti-CD24 antibody binds to the leucine-alanine-proline motif of CD24. Use of an anti-CD24 antibody according to any one of claims 1 to 6.

8. The hepatitis virus is hepatitis B virus (HBV). Use of an anti-CD24 antibody according to any one of claims 1 to 7.

9. The hepatitis virus is hepatitis D virus (HDV). Use of an anti-CD24 antibody according to any one of claims 1 to 8.

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