Effects of GM-CSF addition on humoral modulation and FCGR2b binding in allergy, infection and immunity

Abstract

The present disclosure relates to the prevention of a viral infection or an allergic reaction in a subject who has received a vaccine against the virus or the allergen along with granulocyte-macrophage colony-stimulating factor.

Description

EFFECTS OF GM-CSF ADDITION ON HUMORAL MODULATION AND FCGR2B BINDING IN ALLERGY, INFECTION AND IMMUNITYFIELD

[0001] This disclosure relates to, in part, treatment and / or mitigation of an allergy or an allergic reaction, including the humoral modulation in allergic patients.CROSS-REFERENCE TO RELATED APPLICATION

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 586,445, filed September 29, 2023, the entire contents of which are hereby incorporated by reference in its entirety.DESCRIPTION OF THE XML FILE SUBMITTED ELECTRONICALLY

[0003] The instant application contains a Sequence Listing which has been submitted electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML file, created on September 25, 2024, is named PNR-016PC_127114-5016 PCT_Sequence Listing. xml and is 8,192 bytes in size.BACKGROUND

[0004] The immune system has evolved to protect the host from a multitude of foreign substances including a universe of pathogenic microbes as well as toxic and allergic substances. Beyond the structural (e.g., skin and mucosa) and chemical barriers which protect from infection or an allergen, the immune system can be sim plistically viewed as having two “lines of defense”: innate immunity and adaptive immunity. The innate immunity represents the first line of defense to an intruding pathogen and is antigenindependent (non-specific) defense mechanism that is used by the host immediately or within hours of encountering an antigen. The innate immune response has no immunologic memory and, therefore, it is unable to recognize or “memorize” the same pathogen or allergen should the body be exposed to it in the future. On the other hand, the adaptive immunity is antigen-dependent and antigen-specific and, therefore, involves a lag time between exposure to the antigen and maximal response. The hallmark of adaptive immunity is the capacity for memory which enables the host to mount a more rapid and efficient immune response upon subsequent exposure to the antigen.Importantly, the innate and adaptive immunity are not mutually exclusive mechanisms of host defense, but rather are complementary, with defects in either system resulting in host vulnerability or inappropriate responses. Numerous cells are involved in the innate immune response such as phagocytes (macrophages and neutrophils), dendritic cells, mast cells, basophils, eosinophils, natural killer (NK) cells and innate lymphoid cells. The cells of the adaptive immune system include antigen-specific T cells, which are activated to proliferate through the action of APCs, and B cells which differentiate into plasma cells to produce antibodies or immunoglobulins. See Murphy KM et al. Janeway’s immunobiology. 7th ed. New York: Garland Science; 2007; Turvey SE and Broide DH. J Allergy Clin Immunol. 2010;125(Suppl 2): S24-32; Chaplin DD J Allergy Clin Immunol. 2010. 125(2Suppl 2): S3-23; Bonilla FA and Oettgen HC. J Allergy Clin Immunol. 2010;125(Suppl 2): S33-40; Marshall JS et al. Allergy Asthma Clin Immunol 2018, 14(Suppl 2):49.

[0005] Unlike T cells, B cells can recognize antigens directly, without the need for APCs, through unique antibodies expressed on their cell surface. The principal function of B cells is the production of antibodies against foreign antigens which requires their further differentiation. When activated by foreign antigens to which they have an appropriate antigen specific receptor, B cells undergo proliferation and differentiate into antibodysecreting plasma cells or memory B cells. Memory B cells are “long-lived” survivors of past infection or allergen and continue to express antigen-binding receptors. These cells can be called upon to respond quickly by producing antibodies and eliminating an antigen upon re-exposure. Plasma cells, on the other hand, are relatively short-lived cells that often undergo apoptosis when the inciting agent that induced the immune response is eliminated.

[0006] All antibodies, produced by B cells, are constructed in the same way from paired heavy and light polypeptide chains. Within this general category, however, five different classes of immunoglobulins — IgM, IgD, IgG, IgA, and IgE — can be distinguished by their constant or C regions. More subtle differences, however, are confined to the variable or V regions. Antibody molecules are roughly Y-shaped molecules consisting of three equalsized portions, loosely connected by a flexible tether. Simplistically, each arm of this Y- shaped structure is formed by the association of a light chain with the amino-terminal halfof a heavy chain (Fab fragment), whereas the trunk of the Y is formed by the pairing of the carboxy-terminal halves of the two heavy chains (Fc fragment). The C regions are responsible for binding the effector cells (Fc receptors, FcR), whereas the V regions account for the specificity of antigen binding. See Janeway CA et al. Immunobiology. 5thedition. 2001.

[0007] Of the five immunoglobulin isotypes, immunoglobulin G (IgG) is most abundant in human serum. The four subclasses, lgG1 , lgG2, lgG3, and lgG4, which are highly conserved, differ in their constant region, particularly in their hinges and upper CH2 domains. These regions are involved in binding to both IgG-Fc receptors (FcyR) and C1q. As a result, these structurally distinct subclasses have different effector functions, both in terms of triggering FcyR-expressing cells, resulting in phagocytosis or antibodydependent cell-mediated cytotoxicity, and activating complement. The major classes of antibodies have substantially different biological functions and recognize and neutralize specific pathogens. Typically, lgG1 and lgG3 are potent triggers of effector mechanisms, whereas lgG2 and lgG4 will induce more subtle responses, and only in certain cases. However, these antibodies remain capable of neutralizing virus particles and toxins. Defects or malfunctions in either the innate or adaptive immune response can provoke illness or disease. Such disorders are generally caused by an overactive immune response (known as hypersensitivity reactions), an inappropriate reaction to self (known as autoimmunity) or ineffective immune responses (known as immunodeficiency). Hypersensitivity reactions refer to undesirable responses produced by the normal immune system. There are four types of hypersensitivity reactions: Type I: immediate hypersensitivity, Type II: cytotoxic or antibody-dependent hypersensitivity, Type III: immune complex disease, and Type IV: delayed-type hypersensitivity. See Murphy KM et al. Janeway’s immunobiology. 7th ed. New York: Garland Science; 2007; Turvey SE and Broide DH. J Allergy Clin Immunol. 2010;125(Suppl 2):S24-32; Bonilla FA and Oettgen HC. J Allergy Clin Immunol. 2010;125(Suppl 2):S33^tO; Vidarsson G et al. Front Immunol. 2014; 5:520; Marshall JS et al. Allergy Asthma Clin Immunol 2018, 14(Suppl 2):49.

[0008] Antibody responses to soluble protein antigens and membrane proteins primarily induce lgG1 but are accompanied with lower levels of the other subclasses. BecauselgG1 is normally the most abundant subclass, a lack of IgG 1 seen in a variety of primary and secondary antibody deficiencies, can result in decreased total IgG levels (hypogammaglobulinemia). lgG1 deficiencies, sometimes in combination with other IgG subclass deficiencies, are associated with recurrent infections. Immunoglobulin G- antibody responses to bacterial capsular polysaccharide antigens can be almost completely restricted to lgG2, and lgG2 deficiency may result in the virtual absence of IgG anti-carbohydrate antibodies, although these responses can also be compensated for by enhanced levels of other IgG sub- classes, in particularly by elevated lgG1 and lgG3 levels. Low concentrations of lgG2 often occur in association with a deficiency in lgG4 and / or lgA1 and lgA2. lgG3 antibodies are particularly effective in the induction of effector functions. Being a potent pro-inflammatory antibody, its shorter half-life may function to limit the potential of excessive. Viral infections in general lead to IgG antibodies of the lgG1 and lgG3 subclasses, with lgG3 antibodies appearing first in the course of the infection. See Ferrante A. et al. Pediatr Infect Dis J. 1990 (8 Suppl): S16-24; Jefferis R. et al. Clin Exp Immunol. 1990. 81 (3):357-67; Siber GR. Et al. N Eng J Med. 1980. 303(4): 178-82; Hammarstrom L. et al. Clin Exp Immunol. 1983. 51 (3):600-4; Hammerstrom L. et al. Clin Immunol Immunopathol. 1987. 45(3):461 -70; Vidarsson G et al. Front Immunol. 2014; 5:520.

[0009] lgG4 is a subclass of IgG antibody with a unique molecular feature of (Fragment antigen- binding) Fab-arm exchange, allowing bispecific antigen binding in a mono- valent manner. lgG4 is one of the subclasses detected recently and constitutes about 5% of total IgG, the smallest portion among all IgGs in serum. Although these different subtypes of IgG have more than 90% identical amino acids, they display different immunological effects such as immune complex formation and complement activation. Previous studies have revealed that different IgG subclasses are associated with different antigens. lgG1 and lgG2 subclass is usually associated with the response to bacterial polysaccharides. lgG3 is a potent pro- inflammatory antibody to induce effector function, while lgG4 is for non-microbial allergens. lgG4 can act as a tolerance-inducer to play a protective role under repeated and rapid incremental dosing of allergen exposure in allergen immunotherapy (AIT), supported by allergies in cat raisers and venom desensitization in beekeepers. lgG4 levels may vary significantly in healthy individuals, strongly hinderingits clinical application as a diagnostic tool. Nevertheless, recent findings from molecular structure and clinical investigations have brought new insights into this subset of IgG antibodies in allergic diseases. The production of lgG4 seems to have a very similar pathway to IgE. Both are part of the Th2 immune response, triggered by allergen exposure and produced by a class switch on B cells dependent on IL-4 / IL-13. The immune biological role of lgG4 appears to counteract IgE through different pathways. For example, lgG4 can block IgE binding to specific allergens in serum through competition, though the difference in repertoires may influence the results. Another pathway of countering IgE is the direct inhibition of IgE-mediated mast cell activation. See Aalberse RC et al. Clin Rev Allergy (1983) 1 (2):289-302; Van Milligen FJ et al. Clin Exp Allergy (1995) 25(3):247-51 ; Nirula A et al. Curr Opin Rheumatol (2011 ) 23 (1 ): 119-24; Vidarsson G et al. Front Immunol (2014) 5:520; Qin L et al., Front Immunol. 2022 Oct 25; 13: 1032909.

[0010] Fc receptors (FcRs) are expressed by several hematopoietic cells that recognize Fc regions of several immunoglobulins (Ig) classes and subclasses. FcRs control the humoral and innate immunity which are essential for appropriate responses to infections and prevention of chronic inflammation or auto-immune diseases. Following their crosslinking by immune complexes, FcRs play various roles such as modulation of the immune response by released cytokines or of phagocytosis. The function of antibodies depends on one hand on their ability to recognize antigenic epitopes and, on the other hand, on their dynamic flexibility and their capacity to interact with their cognate FcRs. Several FcRs require the Immunoreceptor Tyrosine- based Activation Motif (ITAM) present in the cytoplasmic tail of the receptor or of associated subunits (FcRy or FcsRip chain) to induce cell signaling. ITAM-mediated functions include phagocytosis, degranulation, antibody-dependent cellular cytotoxicity (ADCC), cytokine, lipid mediator and superoxide production, all of which depend on the cell type and on outside- in signals induced by the ligand. The activation of ITAM-bearing immune receptors can be retro- controlled by ITIM-bearing inhibitory FcRs such as the FcyRIIB. Mkaddem SB et al. 2019. Front Immunol. V10.

[0011] FcyRIIB is the only FcR that contains an inhibitory ITIM motif and serves as a critical negative regulator in immune complex driven reactions. In mice lacking FcyRIIBauto-immune symptoms are exacerbated, and a partial restoration of FcyRIIB expression in B cells rescued mice from developing an SLE-like phenotype. See Bolland S and Ravetch JV. Immunity. (2000) 13:277-85; McGaha TL, et. al. Science. (2005) 307:590- 3; Mkaddem SB et al. 2019. Front Immunol. V10. Recently, there has been a renewed focus on the role of FcyRs in antiviral protection and pathogenesis through antibodydependent enhancement (ADE). ADE has been demonstrated in vitro for IgG antibodies to several diverse viruses, including dengue, influenza, HIV and Ebola virus. Further, Fc- FcyR interactions have been proposed to have a pathogenic role in the context of vaccination against respiratory pathogens such as RSV and influenza. This phenomenon, termed vaccine-associated enhanced respiratory disease (VAERD), was first described in pediatric populations and associated with vaccination with inactivated measles virus or RSV. More recently, this phenomenon has been shown in the context of SARS-CoV2 infections. Important there is recent evidence that enhancing existence of pre-existing IgG antibodies capable of binding to FCGRs may enhance the developing immune response to the pathogen. See Bournas S et al. Nat Rev Immul. 2020 Oct;20(10):633- 643; Boudreau CM. Cell Rep Med. 2023 Mar 21 ;4(3): 100975

[0012] Colony Stimulating Factor (CSF) refers to a family of four glycoproteins that control and coordinate cell production by widely scattered deposits of marrow cells. These include Granulocyte-Macrophage CSF (GM-CSF), Granulocyte colony CSF (G-CSF), Macrophage colony CSF (M-CSF) and multipotential colony-stimulating factor (IL-3). These lymphokines can induce progenitor cells found in the bone marrow to differentiate into specific types of mature blood cells. The particular type of mature blood cell that results from a progenitor cell depends upon the type of CSF present. See Metcalf D. Cancer Immunol Res. 2013, 1(6): 351 -356.

[0013] GM-CSF is a hematologic growth factor that regulates the production, migration, proliferation, differentiation, and function of hematopoietic cells. In response to inflammatory stimuli, GM-CSF is released by various cell types including T lymphocytes, macrophages, fibroblasts, and endothelial cells. GM-CSF then activates and enhances the production and survival of neutrophils, eosinophils, and macrophages. Native GM- CSF is usually produced near the site of action where it can modulate proliferation, differentiation, and survival of hematopoietic progenitor cells. It is present in circulatingblood in only picomolar concentrations (10'1° to 10'12M). See Alexander WS. Int Rev Immunol. 1998, 16:651 -682; Gasson JC. Blood. 1991 , 77:1131-1145; Shannon MF et al. Crit Rev Immunol. 1997, 17:301-323, Barreda DR et al. Dev Comp Immunol. 2004, 28:509-554 and Metcalf D. Immunol Cell Biology. 1987, 65:35-43.

[0014] Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM- CSF) has been approved by the FDA for the treatment of neutropenia, blood dyscrasias and malignancies like leukemia in combination with chemotherapies. In the clinic, GM- CSF used for treatment of neutropenia and aplastic anemia following chemotherapy greatly reduces the risk of infection associated with bone marrow transplantation. Its utility in myeloid leukemia treatment and as a vaccine adjuvant is also well established. See Dorr RT. Clin Therapeutics. 1993. 15(1 ):19-29; Armitage JO. Blood 1998, 92:4491 -4508; Kovacic JC et al. J Mol Cell Cardiol. 2007, 42:19-33; Jacobs PP et al. Microbial Cell Factories 2010, 9:93. Although there are five classes of heterologous protein production platforms, including bacteria, yeasts, plants, insect cells, and mammalian cells, more than 50% of currently marketed biopharmaceuticals are produced in mammalian cell lines. This is in part due to the inability of the remaining four classes to modify glycoproteins with human-like oligosaccharides. This is of importance since protein-bound glycans influence circulation half-life, tissue distribution, biological activity, and immunogenicity. LEUKINE is a yeast-derived recombinant humanized granulocyte-macrophage colony stimulating factor (rhuGM-CSF, sargramostim) and the only FDA approved GM-CSF.

[0015] The interaction between dendritic cells (DC) and naive T cells is the first step in the evolution of an immune response, either tolerogenic or inflammatory. Therefore, the status of DC residing at mucosal sites, such as the airway, has a definitive impact on the character of the ensuing immune response. Important for the nature of the host immune response, GM-CSF can influence characteristics of the DC populations in the mucosa and the interactions between DCs and T cells in the draining lymph nodes that determine proinflam matory versus anti-inflammatory characteristics of the acquired immune response. GM-CSF can have an important role in increasing mucosal defense by regulating DC survival and DC localization during microbial infection. For example, studies from animal models indicate that GM-CSF can have a key role in host defense and regulating inflammation in the intestinal mucosa, and that this is most evident whenintestinal homeostasis is disturbed. Conversely, a relative deficiency of GM-CSF may render an individual more susceptible to the development of acute and / or chronic intestinal inflammation during, for example, intestinal infection or injuries that alter intestinal barrier function. See Egea L et al. Expert Rev Gastroenterol Hepatol. 2010 Dec; 4(6): 723-731. In the absence of pathogenic stimulation, DC serve to regulate immunological homeostasis in the lung; the generation of Th2-associated (allergic) inflammatory responses, which are directed at presumably innocuous antigens, represent a deviation from normal DC function. The dysregulation of DC phenotype leading to the development of allergy might be programmed by genetic pedigree or might be induced by factors released in the airway. Given the pluripotent nature of GM-CSF, it could be used to condition DC to propagate Th2 responses. See Gajewska BU et al. Curr Drug Targets Inflamm Allergy. 2003 Dec;2(4):279-92. The pathogenesis of allergic diseases entails an ineffective tolerogenic immune response to allergens. Regulatory T (Treg) cells have been shown to play a key role in sustaining immune tolerance to allergens. Genetic and immunological evidence also reinforce the idea of a pivotal role for regulatory T (TReg) cells in promoting tolerance to allergens and preventing allergic disorder. See Chatila TA et al. J Clin Invest. 2000; 106:R75-R81 ; Torgerson TR et al. Gastroenterology. 2007; 132:1705-17; Palomares O et al. Eur J Immunol. 2010; 40:1232-40; Jones SM et al. J Allergy Clin Immunol. 2014; 133:318-23; Palomares O et al. Genes Immun. 2014; 15:511-20; Rivas MN and Chatila TA J Allergy Clin Immunol. 2016 Sep; 138(3): 639- 652.

[0016] The rapid spread and unpredictable disease such as COVID caused by SARS- CoV-2 and its variants has clearly illustrated the urgent need for vaccines against pathogens that have the potential to cause pandemics. Specific allergen immunotherapy (AIT) is the only treatment option currently available that has a disease-modifying effect which persists after treatment is stopped. In both these scenarios, a therapeutic that could synergistically act to enhance such a vaccines or therapies could be of immense clinical and economical value. There is further need for therapies that could prevent and / or mitigate immune response to pathogens and allergens.SUMMARY

[0017] Accordingly, in aspects, the present disclosure relates to a method for preventing an infection or an allergy in a subject who has or is receiving a vaccine to a viral pathogen or an allergen, comprising administering an effective amount of a composition comprising granulocyte-macrophage colony-stimulating factor (GM-CSF) to a subject or patient in need thereof.

[0018] In aspects, the present disclosure provides a method for preventing an infection in a subject who is receiving a viral vaccine to the viral pathogen comprising: (a) selecting a patient who has received a vaccination to the infection; and (b) administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent; wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following vaccination to the infection, and to enhance the FcyR2B-binding antibody profile following vaccination to the infection.

[0019] In aspects, the present disclosure provides a method for preventing an allergy in a subject who is receiving an allergen immunotherapy for the allergy, comprising: (a) selecting a patient who has received an immunotherapy to an allergen associated with the allergic reaction; and (b) administering with an effective amount of a composition comprising a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent; wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following immunization to the allergen, and to enhance the FcyR2B-binding antibody profile following immunization to the allergen.

[0020] In aspects, there is provided a method of preventing a viral infection, comprising selecting a patient who has received a vaccination to the infection and administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent, wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following vaccination to the infection.

[0021] In aspects, there is provided a method of modulating or reshaping a humoral immune response to a viral infection in a subject comprising administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent to the subject, wherein the effective amount of GM-CSF is sufficient to enhance the FcyR2B- binding antibody profile following vaccination to the infection.

[0022] In aspects, there is provided a method of preventing an allergic reaction, comprising selecting a patient who is receiving or has received an immunotherapy to an allergen or an allergen immunotherapy and administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent, wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following immunization to the allergen.

[0023] In aspects, there is provided a method of modulating a humoral immune response to an allergen in a subject comprising administering an effective amount of a granulocytemacrophage colony-stimulating factor (GM-CSF) agent to the subject, wherein the effective amount of GM-CSF is sufficient to enhance the FcyR2B-binding antibody profile following immunization to the allergen.

[0024] In embodiments, the present methods cause an increase of the pre-existing Fcy- receptor (FcyR)-binding antibody profile relative to an untreated or pre-treated state.

[0025] In embodiments, the present methods cause an increase in the FcyR2B-binding antibody profile relative to an untreated or pre-treated state.BRIEF DESCRIPTION OF THE DRAWINGS

[0026] FIGURE 1A depicts a randomized, open label clinical trial design.

[0027] FIGURE 1B shows the type of COVID vaccine received by patients in the clinical trial design.

[0028] FIGURE 1C shows the patient vaccination status in the biomarker study.

[0029] FIGURE 2 shows a graph depicting FcGR2B-binding antibody profile in vaccinated patients. For orientation, for each condition on the x-axis (i.e. “Ancestral trimer,” “Delta trimer,” “Gamma P.1 trimer,” “Beta trimer,” and “Alpha trimer”) there are four histograms, which are left to right: “Vaccinated-Placebo vs Unvaccinated-Placebo”, “Vaccinated- Sagramostim vs Unvaccinated-Sagramostim”, “Unvaccinated-Sagramostim vs Unvaccinated-Placebo,” “Vaccinated-Sagramostim vs Vaccinated-Placebo”.

[0030] FIGURE 3 shows a graph depicting the correlation between FcGR2B and viral load clearance.

[0031] FIGURE 4A shows a table depicting the overall symptom score change from baseline in patients with and without treatment with sargramostim.

[0032] FIGURE 4B shows a graph of the symptom scores over 28 days in vaccinated-patients vs. unvaccinated-patients with and without treatment with sargromostim. For orientation, the open red star line depicts the unvaccinated- sargramostim treated patients, the open diamond line depicts the vaccinated- sargramostim treated patients, the colored circle line depicts the unvaccinated-placebo treated patients, the open circle line depicts the vaccinated-placebo treated patients.DETAILED DESCRIPTION

[0033] The present disclosure further relates to, in part, that granulocyte-macrophage colony-stimulating factor (GM-CSF) combined with a viral vaccine is an effective agent for treating and / or preventing a breakthrough viral infection, including, for example, modulating the breadth of the neutralizing antibodies following vaccination to the infection and specifically enhancing the FcyR2B-binding antibody profile following vaccination to the infection.

[0034] The present disclosure relates to, in part, the finding that granulocyte-macrophage colony-stimulating factor (GM-CSF) combined with an allergen immunotherapy to an allergen is an effective agent for allergy prevention, including, for example, modulating the breadth of the neutralizing antibodies following immunization to the allergen and specifically enhancing the FcyR2B-binding antibody profile following immunization to the allergen.

[0035] Accordingly, in aspects, the disclosure provides methods for preventing an infection or an allergy in a patient immunized to the virus or allergy.Viruses and Viral Infections

[0036] In aspects, the disclosure provides methods for preventing a viral infection in a patient immunized to the virus.

[0037] In embodiments, the viral infection is a respiratory viral infection. In some embodiments, the respiratory viral infection is an infection with a coronavirus. In other embodiments, the respiratory viral infection is an infection with an influenza virus.Coronavirus

[0038] The coronavirus is a member of the family Coronaviridae, including betacoronavirus and alphacoronavirus respiratory pathogens that have relatively recently become known to invade humans. The Coronaviridae family includes such betacoronavirus as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS-CoV, Middle East Respiratory Syndrome-Corona Virus (MERS-CoV), HCoV- HKU1 , and HCoV-OC43. Alphacoronavirus includes, e g., HCoV-NL63 and HCoV-229E.

[0039] Coronaviruses invade cells through “spike” surface glycoprotein that is responsible for viral recognition of Angiotensin Converting Enzyme 2 (ACE2), a transmembrane receptor on mammalian hosts that facilitate viral entrance into host cells. Zhou et al., A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature 2020. A new coronavirus infection 2019 (COVID-19), caused by

[0040] SARS-CoV-2 causes severe respiratory distress, and this RNA virus strain has been the cause of the recent outbreak that has been declared a major threat to public health and worldwide emergency. Phylogenetic analysis of the complete genome of SARS-CoV-2 revealed that the virus was most closely related (89.1 % nucleotide similarity) to a group of SARS-like coronaviruses (genus Betacoronavirus, subgenus Sarbecovirus). Wu et al., A new coronavirus associated with human respiratory disease in China. Nature, Feb 3, 2020.

[0041] The complete genome of the SARS-CoV-2 coronavirus (29903 nucleotides, singlestranded RNA) is described in the NCBI database as GenBank Reference Sequence: MN908947. In embodiments, the coronavirus protein is selected from coronavirus spike protein (GenBank Reference Sequence: QHD43416), coronavirus membrane glycoprotein M (GenBank Reference Sequence: QHD43419), coronavirus envelope protein E (GenBank Reference Sequence: QHD43418), and coronavirus nucleocapsid phosphoprotein E (GenBank Reference Sequence: QHD43423).

[0042] The SARS-CoV-2 is an enveloped, single stranded, RNA virus that encodes a “spike” protein, also known as the S protein, which is a surface glycoprotein that mediates binding to a cell surface receptor; an integral membrane protein; an envelope protein, and a nucleocapsid protein. The S protein, comprising S1 subunit and S2 subunit, is a trimeric class I fusion protein that exists in a prefusion conformation that undergoes a structural rearrangement to fuse the viral membrane with the host-cell membrane. See, e.g., Li, F. Structure, Function, and Evolution of Coronavirus Spike Proteins. Annu. Rev. Virol. 3: 237-261 (2016 which is incorporated herein by reference in its entirety. The structure of the SARS-CoV-2 spike protein in the prefusion conformation has been discovered. See Daniel et al., Cryo-EM structure of the SARS-CoV-2 spike in the prefusion conformation. Science, 19 Feb 2020, which is incorporated herein by reference in its entirety.

[0043] In embodiments, the method treats a breakthrough with SARS-CoV-2 variants of concern (VOC), wherein the variant of SARS-CoV-2 are selected from alpha (B.1.1.7), beta (B.1.351 ), gamma (P.1 ), delta (B.1.617.2), epsilon (B.1.427 or B.1.429), eta (B.1.525), iota (B.1.526), kappa (B.1.617.1 ), lambda (C.37), mu (B.1.621 or B.1.621.1 ), omicron (BA.1 , B.1.1.529, BA.2, BA.2.75, BA.3, BA.4 / BA.5, BN.1 BQ.1 , XBB, XBB.1.5, XAK, XAY, XBC), pirola (BA.2.86) and zeta (P.2).Influenza virus

[0044] The influenza virus is a member of the family Orthomyxoviridae, which include enveloped viruses with segmented negative-sense single-strand RNA segments. There are several genera of this family including types A, B, C, D and Thogotovirus, of which, however, only genera A and B are clinically relevant for humans. See Untergruppe AB. Transfus Med Hemother. 2009. 36(1 ):32-39.

[0045] The eight genome segments of influenza A and B viruses are loosely encapsidated by the nucleoprotein. The polymerase complexes consisting of the three polymerase proteins PB1 , PB2, and PA are located at the ends of the nucleocapsids. These helical capsids are encircled by the M1 matrix protein and by a host-derived lipid bilayer envelope in which the virus surface glycoproteins haemagglutinin (HA) and neuraminidase (NA) as well as the M2 matrix protein are embedded. In embodiments, the influenza virus is selected from Type A, Type B, Type C, and Type D influenza.

[0046] In embodiments, the viral infection is pandemic 2009 influenza A (H1 N1 ) or avian influenza A (H5N1 ).Viral Infection sequelae

[0047] In embodiments, the patient has experienced one or more prior infections by or exposures to the virus. In some embodiments, the patient has experienced one prior infection by or exposure to the virus. In some embodiments, the patient has experienced two or more prior infections by or exposures to the virus. In other embodiments, the patient has experienced three or more prior infections by or exposures to the virus. In yet other embodiments, the patient has experienced one or more prior infections by or exposures to the virus of at least about two weeks before the infection.

[0048] In embodiments, the patient has mild or moderate COVID-19.

[0049] In embodiments, the patient is afflicted with one or more of fever, cough, shortness of breath, diarrhea, upper respiratory symptoms, lower respiratory symptoms, pneumonia, and acute respiratory syndrome. In some embodiments, the patient is afflicted with fever, tiredness, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper respiratory symptoms (e.g. sneezing, runny nose, nasal congestion, cough, sore throat), pneumonia, pneumonia respiratory failure, hepatic and renal insufficiency, acute respiratory distress syndrome (ARDS), and a cytokine imbalance by one or more symptoms of a coronavirus infection relative to an uninfected state.

[0050] In embodiments, the patient is elderly and / or afflicted with one or more comorbidities. In embodiments, the patient has one or more characteristics which provide a high risk for progression to severe disease. In some embodiments, the patient has one or more pre-existing medical conditions that reduce an immune response. In some embodiment, the patient is overweight or obese.Viral vaccines

[0051] In embodiments, the patient is vaccinated with a COVID vaccine specific against an infection with SARS-CoV2, optionally selected from tozinameran (COMIRNATY) or elasomeran (mRNA-1273, SPIKEVAX), or a variant thereof.

[0052] In some embodiments, the COVID vaccine is or comprises an mRNA- based vaccine. In embodiments, the mRNA vaccine comprises one or more non- canonical nucleotides, optionally selected from N1 -methylpseudouridine, pseudouridine, and 5-methoxyuridine. In embodiments, the COVID vaccine comprises one or more lipids.

[0053] In some embodiments, the COVID vaccine is or comprises a DNA-based vaccine.

[0054] In some embodiments, the COVID vaccine is or comprises a viral vectorbased vaccine. In embodiments, the viral vector-based vaccine is or comprises an adeno-associated-based vaccine, optionally selected from Ad26.COV2.S or AZD1222 (COVISHIELD, VAXZEVRIA), or a variant thereof.

[0055] In some embodiments, the COVID vaccine is or comprises an inactivated vaccine. In other embodiments, the COVID vaccine is or comprises a live-attenuated vaccine. In embodiments, the COVID vaccine is or comprises a subunit, recombinant, polysaccharide, or conjugate vaccine. In some embodiments, the COVID vaccine comprises a spike surface glycoprotein, membrane glycoprotein M, envelope protein E, and / or nucleocapsid phosphoprotein N, or a fragment thereof. In other embodiments, COVID vaccine comprises a nucleic acid encoding a spike surface glycoprotein, membrane glycoprotein M, envelope protein E, and / or nucleocapsid phosphoprotein N, or a fragment thereof.

[0056] In other embodiments, the patient is vaccinated with an influenza vaccine. In embodiments, the influenza vaccine is or comprises an mRNA-based vaccine. In some embodiments, the influenza vaccine is or comprises a DNA-based vaccine. In some embodiments, the influenza vaccine is or comprises an inactivated vaccine. In some embodiments, the influenza vaccine is or comprises a live-attenuated vaccine.

[0057] In embodiments, the influenza vaccine is or comprises a subunit, recombinant, polysaccharide, or conjugate vaccine. In embodiments, the influenza vaccine comprises an influenza protein, or an antigenic fragment thereof, optionally selected from hemagglutinin (HA) protein, matrix 2 (M2) protein, and neuraminidase, or an antigenic fragment thereof. In other embodiments, the influenza vaccine comprises a nucleic acid encoding an influenza protein, or an antigenic fragment thereof, optionallyselected from hemagglutinin (HA) protein, matrix 2 (M2) protein, and neuraminidase, or an antigenic fragment thereof.Allergens, Allergic Diseases and Allergic Immune Responses

[0058] In aspects, the disclosure provides methods for preventing an allergic reaction in a patient immunized to against the allergen.

[0059] In embodiments, allergic diseases include life-threatening anaphylaxis, food allergies, certain forms of asthma, rhinitis, conjunctivitis, angioedema, urticaria, eczema, eosinophilic disorders, including eosinophilic esophagitis, and drug and insect allergies.

[0060] In embodiments, the allergen is or comprises a protein, peptide, lipoproteins, carbohydrate, or are contained or expressed in cells, or fragments or derivatives of the same.

[0061] In embodiments, the allergens is or comprise plant allergens (e.g. tree pollen), insect sting allergens (e.g., bee sting allergens), animal allergens (e.g., pet allergens, e.g., animal dander or cat Fel d1 antigen), latex allergens, fungal (including mold and mildew) allergens, cosmetic allergens, drug allergens, food allergens (e.g. mustard), dust, arthropod allergens (e.g. crustacean allergens, insect allergens, arachnid allergen), sulphites, sulphur dioxide, metal allergens (e.g. nickel, cobalt, chromium, zinc), insect venom, viruses, or bacteria. In embodiments, the allergen is or comprises an asthma antigen, a hay fever antigen, a hives antigen, an eczema antigen, a plant allergen, an insect sting allergen, an insect allergen, an animal allergen, a fungal allergen, a drug allergen, haptens, small chemicals, a pet allergen, a latex allergen, a mold allergen, a cosmetic allergen, or a food allergen.

[0062] In embodiments, the food allergen is selected from milk allergens, egg allergens, nut allergens (e.g., tree nut allergens (e.g., walnuts, cashews, and the like)), fish allergens, crustacean allergens, soy allergens, legume allergens (e.g., peanut allergen), seed allergens and wheat allergens. In embodiments, the insect sting allergen is selected from allergens that are or are associated with bee stings, wasp stings, hornet stings, yellow jacket stings and the like. In embodiments, the arthropod allergens are selected from insect allergens (e.g., cockroach allergens), crustacean allergens (e.g., lobster,shrimp allergens), and arachnid allergens (e.g., house-dust mites Der p1 antigen) and the like. In embodiments, the drug allergen is selected from allergens that are or are associated with antibiotics, NSAIDs, and anaesthetics. In embodiments, the pollen allergen is selected from grass allergens, tree allergens, weed allergens, and flower allergens. In embodiments, the metal allergen, is selected from nickel, cobalt, chromium, and zinc.

[0063] In embodiments, the allergen induces, or is expected to induce, an undesired immune response in a subject. In embodiments, the undesired immune response is or comprises a Type 1 hypersensitive immune response. In embodiments, the undesired immune response is or comprises allergen-specific antibody production. In embodiments, the undesired immune response is or comprises allergen specific CD4+ T cell proliferation and / or activity. In embodiments, the undesired immune response is or comprises allergen-specific B cell proliferation and / or activity.

[0064] In embodiments, the patient has experienced one or more prior exposures to the allergen. In embodiments, the patient has experienced one or more prior reactions to the allergen. In embodiments, the patient is afflicted with one or more of allergic conditions affecting the nose and eyes (e.g., allergic rhino-conjunctivitis), ears (e.g., allergic otitis media), lungs (e.g., bronchial asthma) as well as for severe insect sting allergy. In embodiments, the patient is afflicted with one or more of fever, tiredness, cough, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper respiratory symptoms (e.g., sneezing, runny nose, nasal congestion, cough, sore throat), itchy and / or painful ears, lower respiratory symptoms, and an immune imbalance relative to a non-allergic state. In embodiments, the patient has mild or moderate allergies or allergic symptoms.GM-CSF

[0065] In embodiments, GM-CSF includes any pharmaceutically safe and effective GM- CSF, or any derivative thereof having the biological activity of GM-CSF. In embodiments, the GM-CSF is recombinant human GM-CSF (rhu GM-CSF), such as sargramostim (LEUKINE). Sargramostim is a biosynthetic, yeast-derived, recombinant human GM- CSF, having of a single 127 amino acid glycoprotein that differs from endogenous humanGM-CSF by having a leucine instead of a proline at position 23. Other natural and synthetic GM-CSFs, and derivatives thereof having the biological activity of or substantially similar to natural human GM-CSF, are also useful.

[0066] In embodiments, the GM-CSF is produced or producible in bacteria, yeasts, plants, insect cells, and mammalian cells. In embodiments, the GM-CSF is produced or producible in Escherichia coli cells. In embodiments, the GM-CSF is produced or producible in yeast cells. In embodiments, the GM-CSF is produced or producible in Chinese hamster ovary cells (CHO). In embodiments, the GM-CSF is not produced in E. coli cells. In embodiments, the GM-CSF is produced in a cell that allows for glycosylation, e.g., yeast or CHO cells.

[0067] In embodiments, the GM-CSF has an amino acid sequence of SEQ ID NO: 1 , or a variant of at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98% identity thereto. In embodiments, the GM-CSF has an amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 3, or a variant of at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98% identity thereto. In embodiments, the GM-CSF is one of molgramostim, regramostim and sargramostim. In embodiments, the GM-CSF is sargramostim.

[0068] Without wishing to be bound by theory, the core of hGM-CSF consists of four helices that pack at angles. Crystal structures and mutagenic analysis of rhGM-CSF (Rozwarski D A et al., Proteins 26:304-13, 1996) showed that, in addition to apolar side chains in the protein core, 10 buried hydrogen bonding residues involve intramolecular hydrogen bonding to main chain atoms that were better conserved than residues hydrogen bonding to other side chain atoms; 24 solvation sites were observed at equivalent positions in the two molecules in the asymmetric unit, and the strongest among these is located in clefts between secondary structural elements. Two surface clusters of hydrophobic side chains are located near the expected receptor binding regions. Mutagenesis of residues on the helix A / helix C face confirmed the importance of certain Glu, Gly, and Gin residues. These residues are therefore not to be substituted in the functional substitution variants of hGM-CSF for use in the present disclosure and these helices are to be retained in a functional fragments or deletion variants of hGM-CSF foruse in this disclosure. Further, in embodiments, one of ordinary skill can reference UniProtKB entry P04141 for structure information to inform the identity of variants.

[0069] The N-terminal helix of hGM-CSF governs high affinity binding to its receptor (Shanafelt A B et al., EMBO J 10:4105-12, 1991 ) Transduction of the biological effects of GM-CSF requires interaction with at least two cell surface receptor components, (one of which is shared with the cytokine IL-5). The above study identified receptor binding determinants in GM-CSF by locating unique receptor binding domains on a series of human-mouse hybrid GM-CSF cytokines. The interaction of GM-CSF with the shared subunit of their high affinity receptor complexes was governed by a very small part of the peptide chains. The presence of a few key residues in the N-terminal a-helix of was sufficient to confer specificity to the interaction.

[0070] In embodiments, the amino acid mutations are amino acid substitutions, such as conservative and / or non-conservative substitutions.

[0071] “Conservative substitutions” may be made, for instance, on the basis of similarity in polarity, charge, size, solubility, hydrophobicity, hydrophilicity, and / or the amphipathic nature of the amino acid residues involved. The 20 naturally occurring amino acids can be grouped into the following six standard amino acid groups: (1 ) hydrophobic: Met, Ala, Vai, Leu, He; (2) neutral hydrophilic: Cys, Ser, Thr; Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that influence chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[0072] As used herein, “conservative substitutions” are defined as exchanges of an amino acid by another amino acid listed within the same group of the six standard amino acid groups shown above. For example, the exchange of Asp by Glu retains one negative charge in the so modified polypeptide. In addition, glycine and proline may be substituted for one another based on their ability to disrupt a-helices.

[0073] As used herein, “non-conservative substitutions” are defined as exchanges of an amino acid by another amino acid listed in a different group of the six standard amino acid groups (1 ) to (6) shown above.

[0074] In embodiments, the substitutions include non-classical amino acids (e.g. selenocysteine, pyrrolysine, N-formylmethionine [3-alanine, GABA and b-Aminolevulinic acid, 4-aminobenzoic acid (PABA), D-isomers of the common amino acids, 2,4- diaminobutyric acid, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, y-Abu, s-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosme, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, [3-alanine, fluoro-amino acids, designer amino acids such as [3 methyl amino acids, C a-methyl amino acids, N a-methyl amino acids, and amino acid analogs in general).

[0075] Modification of the amino acid sequences may be achieved using any known technique in the art e.g., site-directed mutagenesis or PCR based mutagenesis. Such techniques are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Plainview, N.Y., 1989 and Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, New York, N.Y., 1989.

[0076] Without wishing to be bound by theory, the degree of glycosylation of biosynthetic GM-CSFs appears to influence half-life, distribution, and elimination. (Lieschke and Burgess, N. Engl. J. Med. 327:28-35, 1992; Dorr, R. T„ Clin. Then 15:19-29, 1993; Horgaard et al., Eur. J. Hematol. 50:32-36, 1993). In embodiments, the present GM-CSF molecules are glycosylated.Allergen-Specific Therapies

[0077] Multiple allergens introduced by different routes are being utilized either clinically or experimentally with subcutaneous (SCIT) and sublingual (SLIT) allergen-specific immunotherapy (AIT). In embodiments, the patient is treated with an allergen-specific immunotherapy (AIT) based therapy.

[0078] In embodiments, the allergen-specific immunotherapy (AIT) is or comprises a subcutaneous immunotherapy (SCIT) or a sublingual immunotherapy (SLIT). In embodiments, the subcutaneous immunotherapy (SCIT) is or comprises a DNA or mRNA-based therapies. In embodiments, the sublingual immunotherapy (SLIT) is or comprises a tablet or a solution (Drop-SLIT).

[0079] In embodiments, the allergen-specific immunotherapy (AIT) is administered in one or more treatment phases. In embodiments, the AIT treatment phases are or comprises a build-up phase or a maintenance phase.Methods of Treatment or Prevention

[0080] In aspects, the present disclosure relates to a method for preventing a viral pathogen in a subject who is receiving a vaccine for the infection, comprising: administering an effective amount of a composition comprising granulocyte-macrophage colony-stimulating factor (GM-CSF) to a subject or patient in need thereof.

[0081] In aspects, the present disclosure provides a method for preventing an infection in a subject who is receiving a viral vaccine to the viral pathogen comprising: (a) selecting a patient who has received a vaccination to the infection; and (b) administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent; wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following vaccination to the infection, and to enhance the FcyR2B-binding antibody profile following vaccination to the infection.

[0082] In aspects, the present disclosure relates to a method for preventing an allergy in a subject who is receiving an allergen immunotherapy for the allergy, comprising: administering an effective amount of a composition comprising granulocyte-macrophage colony-stimulating factor (GM-CSF) to a subject or patient in need thereof.

[0083] In aspects, the present disclosure provides a method for preventing an allergy in a subject who is receiving an allergen immunotherapy for the allergy, comprising: (a) selecting a patient who has received an immunotherapy to an allergen associated with the allergic reaction; and (b) administering with an effective amount of a composition comprising a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent; wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following immunization to the allergen, and to enhance the FcyR2B-binding antibody profile following immunization to the allergen.

[0084] In embodiments, the subject or patient has received one or more doses of the allergen-specific immunotherapy (AIT) therapy. In embodiments, the patient in the build-up phase has received the AIT therapy once or twice every week. In embodiments, the patient in the maintenance phase has received the AIT therapy every two or four weeks. In embodiments, the patient has received a combination of the AIT therapies.

[0085] In embodiments, the method causes a change in the humoral response relative to an untreated or pre-treated state. In embodiments, the method causes an increase in a humoral response relative to an untreated or pre-treated state. In embodiments, the method causes an increase in an antibody response relative to an untreated or pretreated state. In embodiments, the method causes a change in the kinetics of antibody response relative to an untreated or pre-treated state. In embodiments, the method causes a change in the breadth and / or repertoire of the neutralizing antibody response relative to an untreated or pre-treated state. In embodiments, the method causes a change in the breadth of the neutralizing antibodies which is characterized by an increase in the diversity, specificity, potency, and effector function of pre-existing antibodies relative to an untreated or pre-treated state. In embodiments, the method causes an increase of the pre-existing Fcy-receptor (FcyR)-binding antibody profile relative to an untreated or pre-treated state. In embodiments, the method causes an increase in the FcyR2B-binding antibody profile relative to an untreated or pre-treated state.

[0086] In embodiments, the method prevents or mitigates the development or progression of the infection in the patient. In embodiments, the method prevents or mitigates the development or progression of one or more of fever, cough, shortness of breath, diarrhea, upper respiratory symptoms, lower respiratory symptoms, pneumonia, and acute respiratory syndrome.

[0087] In embodiments, the method causes a decrease in viral load in the subject relative to before treatment. In embodiments, the method reduces or mitigates a likelihood of the subject developing ARDS. In embodiments, the method reduces or mitigates a likelihood of the subject developing lung fibrosis. In embodiments, the method prevents or mitigates the development or progression of the viral infection in a subject including one or more of fever, tiredness, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper respiratory symptoms (e.g. sneezing, runny nose, nasal congestion, cough, sore throat), pneumonia, pneumonia respiratory failure, hepatic andrenal insufficiency, lower respiratory symptoms, acute respiratory distress syndrome (ARDS), and a cytokine imbalance.

[0088] In embodiments, the method prevents or mitigates the development or progression of the allergy in the patient. In embodiments, the method prevents or mitigates the development or progression of one or more of allergic conditions affecting the nose and eyes (e.g., allergic rhino-conjunctivitis), ears (e.g., allergic otitis media), lungs (e.g., bronchial asthma), skin and / or gastrointestinal tract. In embodiments, the method prevents or mitigates the development or progression of one or more of fever, tiredness, cough, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper fever, tiredness, cough, dry cough, aches and pains, shortness of breath and other breathing difficulties, asthma, diarrhea, upper respiratory symptoms (e.g. rhinitis, sneezing, runny nose, nasal congestion, cough, sore throat), conjunctivitis, itchy and / or painful ears, lower respiratory symptoms, nausea, vomiting, diarrhea, urticaria, angio-oedema, atopic dermatitis, eczema, eosinophilic disorders (e.g. eosinophilic esophagitis)and an immune imbalance.

[0089] In embodiments, the method partially inhibits an immune response to reduce or ablate an immune imbalance in the subject. In embodiments, the method reverses, reduces or prevents an immune response to the allergen. In embodiments, the method prevents or mitigates the development or progression of an allergy-related morbidity of the patient. In embodiments, the method prevents or mitigates the development or progression of an immune reaction that requires allergy medication.

[0090] In embodiments, the method prevents or mitigates the development or progression of an infection-related morbidity of the patient. In embodiments, the method prevents or mitigates the development or progression of the infection that requires hospitalization. In embodiments, the method prevents or mitigates the development or progression of the infection that causes death of the subject. In embodiments, the method prevents or mitigates the development or progression of the infection that requires receiving invasive mechanical ventilation. In embodiments, the method prevents or mitigates the development or progression of the infection that requires receiving extracorporeal membrane oxygenation (ECMO). In embodiments, the method prevents or mitigates thedevelopment or progression of the infection that requires receiving non-invasive mechanical ventilation. In embodiments, the method prevents or mitigates the development or progression of the infection that requires use of a high flow oxygen device. In embodiments, the method prevents or mitigates the development or progression of the infection that requires receiving supplemental oxygen. In embodiments, the method prevents or mitigates the development or progression of the infection that requires receiving ongoing medical care.

[0091] In embodiments, the method prevents or mitigates the development or progression of the viral infection or allergic reaction / response in the subject.Pharmaceutically Acceptable Salts and Excipients

[0092] The compositions described herein can possess a sufficiently basic functional group, which can react with an inorganic or organic acid, or a carboxyl group, which can react with an inorganic or organic base, to form a pharmaceutically acceptable salt. A pharmaceutically acceptable acid addition salt is formed from a pharmaceutically acceptable acid, as is well known in the art. Such salts include the pharmaceutically acceptable salts listed in, for example, Journal of Pharmaceutical Science, 66, 2-19 (1977) and The Handbook of Pharmaceutical Salts; Properties, Selection, and Use. P. H. Stahl and C. G. Wermuth (eds.), Verlag, Zurich (Switzerland) 2002, which are hereby incorporated by reference in their entirety.

[0093] Pharmaceutically acceptable salts include, by way of non-limiting example, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, pamoate, phenylacetate, trifluoroacetate, acrylate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene- 2-benzoate, isobutyrate, phenylbutyrate, a-hydroxybutyrate, butyne-1 ,4-dicarboxylate, hexyne-1 ,4-dicarboxylate, caprate, caprylate, cinnamate, glycollate, heptanoate,hippurate, malate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate, phthalate, teraphthalate, propiolate, propionate, phenylpropionate, sebacate, suberate, p-bromobenzenesulfonate, chlorobenzenesulfonate, ethylsulfonate, 2- hydroxyethylsulfonate, methylsulfonate, naphthalene-1 -sulfonate, naphthalene-2- sulfonate, naphthalene-1 ,5-sulfonate, xylenesulfonate, and tartarate salts.

[0094] The term “pharmaceutically acceptable salt” also refers to a salt of the compositions of the present disclosure having an acidic functional group, such as a carboxylic acid functional group, and a base. Suitable bases include, but are not limited to, hydroxides of alkali metals such as sodium, potassium, and lithium; hydroxides of alkaline earth metal such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia, and organic amines, such as unsubstituted or hydroxysubstituted mono-, di-, or tri-alkylamines, dicyclohexylamine; tributyl amine; pyridine; N- methyl, N-ethylamine; diethylamine; triethylamine; mono-, bis-, or tris-(2-OH-lower alkylamines), such as mono-; bis-, or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert- butylamine, or tris-(hydroxymethyl)methylamine, N,N-di-lower alkyl-N-(hydroxyl-lower alkyl)-amines, such as N,N-dimethyl-N-(2-hydroxyethyl)amine or tri-(2- hydroxyethyl)amine; N-methyl-D-glucamine; and amino acids such as arginine, lysine, and the like.

[0095] In embodiments, the compositions described herein are in the form of a pharmaceutically acceptable salt.Pharmaceutical Compositions and Formulations

[0096] In embodiments, the present disclosure pertains to pharmaceutical compositions comprising the compositions, e.g., GM-CSF and / or an additional therapeutic agent, described herein and a pharmaceutically acceptable carrier or excipient. Any pharmaceutical compositions described herein can be administered to a subject as a component of a composition that comprises a pharmaceutically acceptable carrier or vehicle. Such compositions can optionally comprise a suitable amount of a pharmaceutically acceptable excipient so as to provide the form for proper administration.

[0097] In embodiments, pharmaceutical excipients can be liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. The pharmaceutical excipients can be, for example, saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. In addition, auxiliary, stabilizing, thickening, lubricating, and coloring agents can be used. In embodiments, the pharmaceutically acceptable excipients are sterile when administered to a subject. Water is a useful excipient when any agent described herein is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid excipients, specifically for injectable solutions. Suitable pharmaceutical excipients also include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. Any agent described herein, if desired, can also comprise minor amounts of wetting or emulsifying agents, or pH buffering agents. Other examples of suitable pharmaceutical excipients are described in Remington’s Pharmaceutical Sciences 1447-1676 (Alfonso R. Gennaro eds., 19th ed. 1995), incorporated herein by reference.

[0098] The present disclosure includes the described pharmaceutical compositions (and / or additional therapeutic agents) in various formulations. Any inventive pharmaceutical composition (and / or additional therapeutic agents) described herein can take the form of solutions, suspensions, emulsion, drops, tablets, pills, pellets, capsules, capsules containing liquids, gelatin capsules, powders, sustained-release formulations, suppositories, emulsions, aerosols, sprays, suspensions, lyophilized powder, frozen suspension, desiccated powder, or any other form suitable for use. In embodiments, the composition is in the form of a capsule. In embodiments, the composition is in the form of a tablet. In embodiments, the pharmaceutical composition is formulated in the form of a soft-gel capsule. In a further embodiment, the pharmaceutical composition is formulated in the form of a gelatin capsule. In embodiments, the pharmaceutical composition is formulated as a liquid.

[0099] Where necessary, the inventive pharmaceutical compositions (and / or additional therapeutic agents) can also include a solubilizing agent. Also, the agents can be delivered with a suitable vehicle or delivery device as known in the art. Combination therapies outlined herein can be co-delivered in a single delivery vehicle or delivery device.

[0100] The formulations comprising the inventive pharmaceutical compositions (and / or additional therapeutic agents) of the present disclosure may conveniently be presented in unit dosage forms and may be prepared by any of the methods well known in the art of pharmacy. Such methods generally include the step of bringing the therapeutic agents into association with a carrier, which constitutes one or more accessory ingredients. Typically, the formulations are prepared by uniformly and intimately bringing the therapeutic agent into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product into dosage forms of the desired formulation (e.g., wet, or dry granulation, powder blends, etc., followed by tableting using conventional methods known in the art).

[0101] In embodiments, any pharmaceutical compositions (and / or additional therapeutic agents) described herein is formulated in accordance with routine procedures as a composition adapted for a mode of administration described herein.

[0102] Routes of administration include, for example: oral, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, sublingual, intranasal, intracerebral, intravaginal, transdermal, rectally, by inhalation, or topically. Administration can be local or systemic. In embodiments, the administering is affected orally. In embodiments, the administration is by parenteral injection. The mode of administration can be left to the discretion of the practitioner and depends in-part upon the site of the medical condition. In most instances, administration results in the release of any agent described herein into the bloodstream.

[0103] In embodiments, the GM-CSF (and / or other additional therapeutic agents) is administered via a subcutaneous route.

[0104] In embodiments, the GM-CSF (and / or other additional therapeutic agents) is administered via an oral (e.g., sublingual) route.

[0105] In embodiments, the GM-CSF (and / or additional therapeutic agents) is administered via an intranasal route. In embodiments, the GM-CSF (and / or additional therapeutic agents) is administered by inhalation. Without wishing to be bound by theory, inhalation of sargramostim produces low systemic exposure in the patient.

[0106] In embodiments, the GM-CSF (and / or additional therapeutic agents) is administered to the lung.

[0107] In embodiments, the GM-CSF (and / or additional therapeutic agents) is administered via aerosol or nebulizer.

[0108] In embodiments, the aerosol or nebulizer is selected from liquid nebulization, dry powder dispersion and meter-dose administration. In embodiments, the aerosol or nebulizer is selected from jet flow or mesh vibrating.

[0109] In embodiments, the GM-CSF (and / or additional therapeutic agents) is administered by inhalation and mediates local and / or peripheral cellular responses. For instance, in embodiments, the GM-CSF (and / or additional therapeutic agents) is administered by inhalation and mediates an increase in lymphocytes and / or eosinophils in the peripheral blood.

[0110] Accordingly, in embodiments, there is provided a method of modulating peripheral immune cells to cause an anti-infective or anti-allergy effect through local administration (e.g., inhalation).

[0111] In embodiments, the pharmaceutical compositions (and / or additional therapeutic agents) described herein are formulated in accordance with routine procedures as a composition adapted for oral administration. Compositions for oral delivery can be in the form of tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups, or elixirs, for example. Orally administered compositions can comprise one or more agents, for example, sweetening agents such as fructose, aspartame, or saccharin; flavoring agents such as peppermint, oil of Wintergreen, or cherry; coloring agents; and preserving agents, to provide a pharmaceutically palatable preparation. Moreover, where in tablet or pill form, the compositions can be coated to delay disintegration and absorption in the gastrointestinal tract thereby providing a sustained action over an extended period of time. Selectively permeable membranes surrounding an osmotically active driving any pharmaceutical compositions (and / or additional therapeutic agents) described herein are also suitable for orally administered compositions. In these latter platforms, fluid from the environment surrounding the capsule is imbibed by the driving compound, which swells to displace theagent or agent composition through an aperture. These delivery platforms can provide an essentially zero order delivery profile as opposed to the spiked profiles of immediate release formulations. A time-delay material such as glycerol monostearate or glycerol stearate can also be useful. Oral compositions can include standard excipients such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, and magnesium carbonate. In embodiments, the excipients are of pharmaceutical grade. Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, tragacanth, etc., and mixtures thereof.

[0112] Dosage forms suitable for parenteral administration (e.g., intravenous, intramuscular, intraperitoneal, subcutaneous, and intra-articular injection and infusion) include, for example, solutions, suspensions, dispersions, emulsions, and the like. They may also be manufactured in the form of sterile solid compositions (e.g., lyophilized composition), which can be dissolved or suspended in sterile injectable medium immediately before use. They may contain, for example, suspending or dispersing agents known in the art. Formulation components suitable for parenteral administration include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetates, citrates or phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose.

[0113] For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). The carrier should be stable under the conditions of manufacture and storage and should be preserved against microorganisms. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol), and suitable mixtures thereof.

[0114] The compositions provided herein, alone or in combination with other suitable components, can be made into aerosol formulations (i.e., “nebulized”) to beadministered via inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.

[0115] Any inventive pharmaceutical compositions (and / or additional therapeutic agents) described herein can be administered by controlled-release or sustained-release means or by delivery devices that are well known to those of ordinary skill in the art. Examples include, but are not limited to, those described in U.S. Patent Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595; 5,591 ,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; and 5,733,556, each of which is incorporated herein by reference in its entirety. Such dosage forms can be useful for providing controlled- or sustained-release of one or more active ingredients using, for example, hydropropyl cellulose, hydropropylmethyl cellulose, polyvinylpyrrolidone, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or a combination thereof to provide the desired release profile in varying proportions. Suitable controlled- or sustained-release formulations known to those skilled in the art, including those described herein, can be readily selected for use with the active ingredients of the agents described herein. The disclosure thus provides single unit dosage forms suitable for oral administration such as, but not limited to, tablets, capsules, gelcaps, and caplets that are adapted for controlled- or sustained-release.

[0116] Controlled- or sustained-release of an active ingredient can be stimulated by various conditions, including but not limited to, changes in pH, changes in temperature, stimulation by an appropriate wavelength of light, concentration or availability of enzymes, concentration or availability of water, or other physiological conditions or compounds.

[0117] In embodiments, a controlled-release system can be placed in proximity of the target area to be treated, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Other controlled-release systems discussed in the review by Langer, 1990, Science 249: 1527-1533) may be used.

[0118] Pharmaceutical formulations preferably are sterile. Sterilization can be accomplished, for example, by filtration through sterile filtration membranes. Where thecomposition is lyophilized, filter sterilization can be conducted prior to or following lyophilization and reconstitution.Administration and Dosage

[0119] It will be appreciated that the actual dose of the composition to be administered according to the present disclosure will vary according to the particular dosage form, and the mode of administration. Many factors that may modify the action of the composition (e.g., body weight, gender, diet, time of administration, route of administration, rate of excretion, condition of the subject, drug combinations, genetic disposition, and reaction sensitivities) can be taken into account by those skilled in the art. Administration can be carried out continuously or in one or more discrete doses within the maximum tolerated dose. Optimal administration rates for a given set of conditions can be ascertained by those skilled in the art using conventional dosage administration tests.

[0120] In embodiments, the GM-CSF is administered at a total dose of about 125 pg, about 150 pg, or about 200 pg, or about 250 pg, or about 300 pg, or about 350 pg. In embodiments, the GM-CSF is administered at a dose of about 125 pg, about 150 pg, or about 200 pg, or about 250 pg, or about 300 pg, or about 350 pg, once daily. In embodiments, the GM-CSF is administered at a total dose of about 250 pg, once daily.

[0121] In embodiments, the GM-CSF is administered at a dose of about 125 pg, about 150 pg, or about 200 pg, or about 250 pg, or about 300 pg, or about 350 pg, once daily. In embodiments, the GM-CSF is sargramostim, administered at a dose of about 250 pg, once daily. In embodiments, the GM-CSF is sargramostim, administered at a dose of about 250 pg, once daily, for at least 5 consecutive days.Combination Therapy and Additional Therapeutic Agents

[0122] In embodiments, the method further comprises administering one or more additional therapeutic agents to treat the viral infection or an allergic reaction / response.

[0123] In embodiments of the disclosure “treating” includes administration of the present combination to an individual who is at risk for potential exposure, or who has been exposed to or a source of the virus. “Treating” means, for example, administration of aviral vaccine (e.g., COVID vaccine or influenza vaccine or another vaccine for a virus) along with GM-CSF, including the combination of the present disclosure, to an individual who tested positive for the viral infection but has not yet shown mild or moderate symptoms, and administration of a combination of the present disclosure to an individual who shows or has shown viral infection symptoms.

[0124] In embodiments of the disclosure “treating” includes administration of the inventive combination to an individual who is at risk for potential exposure, or who has been exposed to or a source of the allergen. “Treating” means, for example, administration of an allergen-specific therapy (e.g., specific allergen immunotherapy (AIT)) along with GM-CSF, including the combination of the present disclosure, to an individual who tested positive for a specific allergen but has not yet shown allergy symptoms, and administration of a combination of the present disclosure to an individual who shows or has shown allergy symptoms.

[0125] In embodiments, the pharmaceutical composition of the present disclosure is co-administered in conjunction with additional agent(s). Co-administration can be simultaneous or sequential.

[0126] In embodiments, the additional therapeutic agent and the GM-CSF of the present disclosure are administered to a subject simultaneously. The term “simultaneously” as used herein, means that the additional therapeutic agent and the GM- CSF are administered with a time separation of no more than about 60 minutes, such as no more than about 30 minutes, no more than about 20 minutes, no more than about 10 minutes, no more than about 5 minutes, or no more than about 1 minute. Administration of the additional therapeutic agent and the GM-CSF can be by simultaneous administration of a single formulation (e.g., a formulation comprising the additional therapeutic agent and the GM-CSF composition) or of separate formulations (e.g., a first formulation including the additional therapeutic agent and a second formulation including the GM-CSF composition).

[0127] Co-administration does not require the therapeutic agents to be administered simultaneously, if the timing of their administration is such that the pharmacological activities of the additional therapeutic agent and the GM-CSF overlap intime, thereby exerting a combined therapeutic effect. For example, the additional therapeutic agent and the targeting moiety, the GM-CSF composition can be administered sequentially. The term “sequentially” as used herein means that the additional therapeutic agent and the GM-CSF are administered with a time separation of more than about 60 minutes. For example, the time between the sequential administration of the additional therapeutic agent and the GM-CSF can be more than about 60 minutes, more than about 2 hours, more than about 5 hours, more than about 10 hours, more than about 1 day, more than about 2 days, more than about 3 days, more than about 1 week apart, more than about 2 weeks apart, or more than about one month apart. The optimal administration times will depend on the rates of metabolism, excretion, and / or the pharmacodynamic activity of the additional therapeutic agent and the GM-CSF being administered. Either the additional therapeutic agent or the GM-CSF composition may be administered first.

[0128] Co-administration also does not require the therapeutic agents to be administered to the subject by the same route of administration. Rather, each therapeutic agent can be administered by any appropriate route, for example, parenterally or non- parenterally.

[0129] In embodiments, the GM-CSF described herein acts synergistically when co-administered with another therapeutic agent. In embodiments, the targeting moiety, the GM-CSF composition, and the additional therapeutic agent may be administered at doses that are lower than the doses employed when the agents are used in the context of monotherapy.

[0130] In embodiments, the additional therapeutic agent is selected from nirmatrelvir / ritonavir, remdesivir; favipiravir; galidesivir; prezcobix; lopinavir; and / or ritonavir; and / or arbidol lopinavir / ritonavir; and / or ribavirin; and / or IFN-beta; xiyanping; anti-VEGF-A; fingolimod; carrimycin; hydroxychloroquine; darunavir and cobicistat; methylprednisolone; brilacidin; leronlimab; thalidomide, bamlanivimab, casirivimab, and imdevimab.

[0131] In embodiments, the additional therapeutic agent is nirmatrelvir / ritonavir (PAXLOVID). In embodiments, the additional therapeutic agent is an antibody directedagainst an antigen of the virus. In embodiments, the additional therapeutic agent is a monoclonal antibody directed against a coronavirus antigen. In embodiments, the additional therapeutic agent is a monoclonal antibody directed against an influenza antigen. In some embodiments, the additional therapeutic agent is a steroid, optionally selected from dexamethasone or prednisone.

[0132] In embodiments, the additional therapeutic agent is selected from an over the counter (OTC) corticosteroid, a prescription corticosteroid, an over the counter (OTC) antihistamine, a prescription antihistamine, over the counter (OTC) decongestant, a prescription decongestant, mast-cell stabilizers, leukotriene inhibitors, biological medications and / or emergency epinephrine injections.

[0133] In embodiments, the additional therapeutic agent is selected from pill, liquids, nasal sprays and / or drops, throat sprays and / or drops, and eye drops.

[0134] In some embodiments, the method prevents or mitigates the development or progression of the infection that requires receiving an additional therapeutic agent.

[0135] In some embodiments, the method prevents or mitigates the development or progression of the allergy that requires receiving an additional therapeutic agent (for e.g., an allergy medication).Methods of Detecting Therapeutic Effect

[0136] In some embodiments, there is provided a method for detecting the likelihood of successful treatment or response to treatment with the present disclosure for an allergy or an allergic reaction in a patient. In embodiments, there is provided a method for detecting the likelihood of successful treatment or response to treatment with the present disclosure for a viral infection in a patient.

[0137] In embodiments, the allergic response or viral infection is assayed or assayable in a biological sample from the patient (e.g., blood, respiratory fluid (e.g., from an oropharyngeal (OP) or nasopharyngeal (NP) swab, sputum, a nasal secretion), saliva or stool.

[0138] In embodiments, the present disclosure includes monitoring the viral load or allergy symptoms during the course of treatment. In embodiments, the present disclosure causes a decrease in the viral load in the patient relative to an untreated or pre-treated state. In embodiment, the monitoring of the viral load is assayed or assayable using a measurement of a viral nucleic acid or protein. In some the monitoring of the viral load is assayed or assayable using PCR. In other embodiments, the monitoring of the viral load is assayed or assayable using a rapid test. In embodiments, the monitoring of the viral load is monitoring of viral load is assayed or assayable using a lateral flow antigen test.

[0139] In embodiments, there is provided a method for detecting the likelihood of successful treatment or response to the present treatment for a viral infection in a patient. In embodiments, there is provided a method for detecting the likelihood of successful treatment or response to treatment with the present disclosure for a viral infection in a patient or subject. In embodiment, the breakthrough viral infection is assayed or assayable using a measurement of a viral nucleic acid or protein. In some the breakthrough viral infection is assayed or assayable using PCR. In other embodiments, the breakthrough viral infection is assayed or assayable using a rapid test. In embodiments, the breakthrough viral infection is assayed or assayable using a lateral flow antigen test.

[0140] In embodiments, there is provided a method for detecting the likelihood of successful treatment or response to treatment with the present disclosure for allergy or an allergic response in a patient or subject by clinical (e.g., skin prick test or patch test) and / or patient-self assessments (e.g., allergic response or symptoms). In embodiments, the allergic response or immune response to an allergen is assayed or assayable using skin patch test and / or a skin prick test. In embodiments, the allergic response is assayed or assayable using a measurement of a nucleic acid or protein. In embodiments, the allergic response is assayed or assayable using PCR.

[0141] In embodiments, the present disclosure causes a decrease in the allergy immune response in the patient relative to an untreated or pre-treated state.

[0142] In embodiments, the present disclosure restores the immune imbalance and / or restores homeostasis in the patient relative to an untreated or pre-treated state.

[0143] In embodiments, the present disclosure causes a decrease in an allergic reaction, and therefore this reduction is used as a biomarker for treatment. In embodiments, the present disclosure causes a decrease in the reaction to a skin prick or patch test, and therefore this reduction is used as a biomarker for treatment. For instance, a decrease in an allergic reaction or skin reaction to a skin prick or patch test directs towards discontinuation of administration of GM-CSF or a reduction in dosage of GM- CSF.

[0144] In embodiments, a worsening of symptoms in the patient with an allergic reaction at any timepoint directs toward continued administration of the GM-CSF. In embodiments, a decrease in an allergic reaction directs towards discontinuation of administration of GM-CSF or a reduction in dosage of GM-CSF.

[0145] In embodiments, the method prevents or mitigates the development or progression of the allergic response that requires receiving an additional therapeutic agent.

[0146] In embodiments, the present disclosure causes a decrease in viral symptoms in the patient, and therefore this reduction is used as a biomarker for treatment. In embodiments, the present disclosure causes a decrease in the amount of virus in a patient, and therefore this reduction is used as a biomarker for treatment. For instance, a decrease in viral load and / or viral symptoms directs towards discontinuation of administration of GM-CSF or a reduction in dosage of GM-CSF.

[0147] In embodiments, a worsening of symptoms and / or an increase in the viral load in the patient with a viral infection at any timepoint directs toward continued administration of the GM-CSF. In embodiments, a decrease in a viral load and / or viral symptoms directs towards discontinuation of administration of GM-CSF or a reduction in dosage of GM-CSF

[0148] In embodiments, the method prevents or mitigates the development or progression of the viral infection that requires receiving an additional therapeutic agent.Uses of Molgramostim or Regramostim

[0149] In embodiments, the present disclosure pertains to the use of molgramostim or regramostim in a method of treating or preventing a viral infection or an allergic reaction, by administering via inhalation and, optionally, modulating an immune response in the patient.

[0150] In embodiments, such molgramostim is used in a method of treating or preventing an immune response to a virus or a specific allergen. In embodiments, such regramostim is used in a method of treating or preventing an immune response to a virus or a specific allergen.Sequences

[0151] SEQ ID NO: 1 is wild type GM-CSF:

[0152] APARSPSPSTQPWEHVNAIQEAPRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKE NLKDFLLVIPFDCWEPVQE.

[0153] SEQ ID NO: 2 is sargramostim:

[0154] APARSPSPSTQPWEHVNAIQEALRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKE NLKDFLLVIPFDCWEPVQE.

[0155] SEQ ID NO: 3 is molgramostim:

[0156] APARSPSPSTQPWEHVNAIQEARRLLNLSRDTAAEMNETVEVISEMFDLQEPTCLQTRLELYKQGLRGSLTKLKGPLTMMASHYKQHCPPTPETSCATQIITFESFKE NLKDFLLVIPFDCWEPVQE.Definitions

[0157] The following definitions are used in connection with the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of skill in the art to which this disclosure belongs.

[0158] An “effective amount,” when used in connection with an agent effective for the treatment of a viral infection is an amount that is effective for treating or mitigating a response to a virus.

[0159] An “effective amount,” when used in connection with an agent effective for the treatment of an allergy or an allergic response is an amount that is effective for treating or mitigating a response to an allergen.

[0160] As used herein, “a,” “an,” or “the” can mean one or more than one. Further, the term “about” when used in connection with a referenced numeric indication means the referenced numeric indication plus or minus up to 10% of that referenced numeric indication. For example, the language “about 50” covers the range of 45 to 55.

[0161] As referred to herein, all compositional percentages are by weight of the total composition, unless otherwise specified. As used herein, the word “include,” and its variants, is intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that may also be useful in the materials, compositions, devices, and methods of this technology. Similarly, the terms “can” and “may” and their variants are intended to be non-limiting, such that recitation that an embodiment can or may comprise certain elements or features does not exclude other embodiments of the present technology that do not contain those elements or features.

[0162] Although the open-ended term “comprising,” as a synonym of terms such as including, containing, or having, is used herein to describe and claim the invention, the present invention, or embodiments thereof, may alternatively be described using alternative terms such as “consisting of” or “consisting essentially of.”

[0163] This disclosure is further illustrated by the following non-limiting examples.EXAMPLESExample 1 : Design of Clinical Trial of Sarqramostim (LEUKINE) for Treatment of COVID- 19

[0164] A prospective, randomized, open-label, interventional study to investigate the efficacy of sargramostim (LEUKINE) in vaccinated and unvaccinated patient who tested positive for COVID-19 patients was conducted (FIGURE 1 A). 600 patients enrolledin the clinical trial had mild to moderate COVID-19 and were not hospitalized. Most patients in the study (82.5%) were unvaccinated for COVID-19 or had not participated in a COVID-19 vaccination trial. There was no significant difference between treatment groups in the percentage of vaccinated patients. Treatment groups were well-balanced for mean age, sex, race, ethnicity, and body mass index (BMI). The Biomarker Cohort patients were comparable to the overall study population in terms of demographic and baseline disease characteristics. In the Biomarker Cohort patients, 28% of were vaccinated for COVID-19 or had participated in a COVID-19 vaccination trial (FIGUREI B). Of these vaccinated Biomarker Cohort patients, most had been vaccinated with an mRNA COVID-19 vaccine (sargramostim-treated = 11 , placebo-treated = 10) (FIGUREIC)Example 2: Clinical Results

[0165] The Fc receptor Luminex array used the fluorescently coded microspheres to capture up to 500 antigen specificities simultaneously and profile the effector capacity of each antigen specificity by determining the ability of these antigen-specific antibodies to interact with Fc receptors. The target antigens, as well as a positive control antigen (influenza H1 (A / California / 7 / 2009)) and a negative control antigen (Ebola virus gp), was covalently coupled to beads via primary amine conjugation or via biotin:streptavidin interactions. The beads were incubated with diluted serum, allowing “on-bead” affinity purification of antigen-specific antibodies. The bound antigen-specific antibodies were subsequently probed with tetramerized recombinant Fc receptors (FCGR2A 131 H / R, FCGR2B, FCGR3A 158V / F, and FCGR3B), and analyzed using Luminex. The data was reported as the median fluorescence intensity of PE for a specific bead channel. Each sample was run in biological duplicate, and the mean value of the duplicates was reported.

[0166] Random forest classifier models were designed to discriminate treatment and vaccination status and random forest regressors models were designed to map humoral features to viral load. The performance of the models was validated using a rigorous k-fold cross-validation and permutation testing framework. Cross-validation is a widely used strategy in machine learning to evaluate the performance of the model withdata held out, and during k-fold cross-validation, the models were built using subsets of the data and then tested on the remainder of the data to gain an unbiased estimate of model performance. Permutation testing generates suitable “negative control” models in a matched cross-validation framework to assess the significance of the observed model performance. Feature importance estimates were established by permutation of out-of- bag predictor observations within each fold. In essence, without wishing to be bound by theory, the importance of a feature corresponds to the drop in accuracy when the information of that feature is removed from the model.

[0167] Computational models of sargramostim and vaccination status highlight, inter alia, that FcGR2B is important in a synergistic immune response against COVID- specific antigens. Serum FcGR2B-binding antibody levels were assessed in 99 patients of the biomarker cohort (Sargramostim n=48; Placebo n=51 ). As shown in FIGURE 2, Sargramostim enhances FcGR2B-binding antibody profile (shown in AUC) in vaccinated patients. For each trimer, enrichment by Sargramostim to FcGR2B is enhanced by vaccination (compare third and fourth bars, i.e., “Unvaccinated-Sagramostim vs Unvaccinated-Placebo” and “Vaccinated-Sagramostim vs Vaccinated-Placebo”.). For each trimer, enrichment by vaccination to FcGR2B is enhanced by Sargramostim (compare first and second bars, i.e., “Vaccinated-Placebo vs Unvaccinated-Placebo” and “Vaccinated-Sagramostim vs Unvaccinated-Sagramostim” ) (FIGURE 2). This pair of enrichments indicates that there is underlying synergy between vaccination and Sargramostim. Note that the area under the curve (AUC) is a measure of the association between two subsections of the cohort, vaccinated-sargramostim versus vaccinated- placebo. An AUC> 0.6 indicates that FcGR2B binding was higher in the vaccinated- sargramostim arm whereas an AUC < 0.4 indicates that either feature was higher in the vaccinated-placebo arm. AUCs between 0.4 and 0.6 are indicative of no difference.

[0168] Assay measurements were baseline corrected as Day 14-Day 1 (baseline). The color bar denotes how much the viral load has decreased per subject in by Day 14 from baseline (Day 1 ). Change is viral load intensity is shown with colors, where blue corresponds to increasing viral load clearance and orange is no change in viral load. FIGURE 3 depicts the analysis of vaccinated patients only, with circles and triangles corresponding to the Sargramostim and the placebo groups respectively. Across allSARS-CoV-2 antigens, subjects with poor viral load clearance (orange) tend to be in the vaccinated-placebo group and show a lower FcGR2B binding. However, higher viral clearance (blue) in the vaccinated-sargramostim group is associated with greater FcGR2B binding. These data show that FcGR2B binding is a feature that is significantly antigen-agnostic in its ability to correlate the claim of synergy between Sargramostim and COVID-19 vaccination (FIGURE 3).

[0169] Symptoms were evaluated and scored in all (vaccinated and unvaccinated) patients who received sargramostim vs. placebo. Enrollment of COVID-vaccinated patients was limited to 100 patients and most had received a mRNA COVID vaccine. To evaluate the change in symptoms during the study, patients completed an electronic symptom score questionnaire at approximately the same time every day up to day 28. The questionnaire assessed 14 common symptoms of COVID-19 (i.e., cough, headache, body aches, etc.) with patients rating each symptom over the prior 24 hours (0=none, 1 =mild, 2=moderate, 3=severe). The overall symptom score was calculated as a sum of the individual scores from the 14 questions each day. Data analyzed at day 7, 14, and day 28. Importantly, the sargramostim cohort had a greater reduction in overall symptom score than the placebo cohort (FIGURE 4A) regardless of vaccination status. This effect was more pronounced in the vaccinated-sargramostim cohort / arm and all vaccinated patients had a lower overall symptom score at baseline than unvaccinated patients (FIGURE 4B). Vaccinated patients also had a lower baseline overall symptom score. The sargramostim arm had a greater reduction in overall symptom score than the placebo arm over 28 days. The effect of sargramostim on patients’ overall symptom score is observed from day 7 onwards.EQUIVALENTS

[0170] Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically herein. Such equivalents are intended to be encompassed in the scope of the following claims.INCORPORATION BY REFERENCE

[0171] All patents and publications referenced herein are hereby incorporated by reference in their entireties.

[0172] As used herein, all headings are simply for organization and are not intended to limit the disclosure in any manner. The content of any individual section may be equally applicable to all sections.

Claims

CLAIMSWhat is claimed is:1 . A method of preventing a viral infection, comprising:(a) selecting a patient who has received a vaccination to the infection; and(b) administering an effective amount of a granulocyte-macrophage colonystimulating factor (GM-CSF) agent; wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following vaccination to the infection.

2. A method of modulating or reshaping a humoral immune response to a viral infection in a subject comprising administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent to the subject; wherein the effective amount of GM-CSF is sufficient to enhance the FcyR2B-binding antibody profile following vaccination to the infection.

3. A method of preventing an allergic reaction, comprising:(a) selecting a patient who is receiving or has received an immunotherapy to an allergen or an allergen immunotherapy; and(b) administering an effective amount of a granulocyte-macrophage colonystimulating factor (GM-CSF) agent; wherein the effective amount of GM-CSF is sufficient to modulate the breadth of the neutralizing antibodies following immunization to the allergen.

4. A method of modulating a humoral immune response to an allergen in a subject comprising administering an effective amount of a granulocyte-macrophage colony-stimulating factor (GM-CSF) agent to the subject; wherein the effective amount of GM-CSF is sufficient to enhance the FcyR2B-binding antibody profile following immunization to the allergen.

5. The method of any one of the claims 1 to 4, wherein the administration of the composition in subjects causes a change in the humoral response relative to an untreated or pre-treated state.

6. The method of any one of the claims 1 to 5, wherein the method causes an increase in the humoral repertoire relative to an untreated or pre-treated state.

7. The method of any one of the claims 1 to 6, wherein the method causes a change in the kinetics of antibody response relative to an untreated or pre-treated state.

8. The method of any one of the claims 1 to 7, wherein the method causes a change in the breadth and / or repertoire of the neutralizing antibody response relative to an untreated or pre-treated state.

9. The method of claim 8, wherein the change in the breadth of the neutralizing antibodies is characterized by an increase in the diversity, specificity, potency, and effector function of pre-existing antibodies relative to an untreated or pre-treated state.

10. The method of any one of the claims 1 to 9, wherein the method causes an increase of the pre-existing Fcy-receptor (FcyR)-binding antibody profile relative to an untreated or pre-treated state.11 . The method of claim 10, wherein the method causes an increase in the FcyR2B- binding antibody profile relative to an untreated or pre-treated state.

12. The method of claim 1 or claim 2, wherein the viral infection is a respiratory viral infection.

13. The method of claim 1 or claim 2, wherein the viral infection is an infection with a coronavirus.

14. The method of claim 13, wherein the coronavirus is selected from:(i) a betacoronavirus, optionally selected from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS-CoV, Middle East respiratory syndromecorona virus (MERS-CoV), HCoV-HKU1 , and HCoV-OC43; and(ii) an alphacoronavirus, optionally selected from HCoV-NL63 and HCoV-229E.

15. The method of claim 13 or claim 14, wherein the coronavirus is SARS-CoV-2 and / or the subject is afflicted with COVID-19.

16. The method of claim 15, wherein the SARS-CoV-2 is a variant of SARS-CoV-2.

17. The method of claim 16, wherein the variant of SARS-CoV-2 is selected from alpha (B.1.1.7), beta (B.1.351 ), gamma (P.1 ), delta (B.1.617.2), epsilon (B.1.427 or B.1.429), eta (B.1.525), iota (B.1.526), kappa (B.1.617.1 ), lambda (C.37), mu (B.1.621 or B.1.621.1 ), omicron (BA.1 , B.1 .1 .529, BA.2, BA.2.75, BA.3, BA.4 / BA.5, BN.1 , BQ.1 , CH.1.1 , XBB, XBB.1.5, XBB.1.9.1*, XBB.1.9.2*, XBB.1.16*, XBB2.3, XAK, XAY, XBC, FE.1 *), pirola (BA.2.86) and zeta (P.2).

18. The method of any one of the claims 13 to 17, wherein the subject has mild or moderate COVID-19.

19. The method of claim 1 or claim 2, wherein the viral infection is an infection with an influenza virus.

20. The method of claim 19, wherein the influenza virus is selected from Type A, Type B, Type C, and Type D influenza.

21. The method of claim 19 or claim 20, wherein the viral infection is pandemic 2009 influenza A (H1 N1 ) or avian influenza A (H5N1 ).

22. The method of any of the claims 13 to 21 , wherein the subject is afflicted with one or more of fever, cough, shortness of breath, diarrhea, upper respiratory symptoms, lower respiratory symptoms, pneumonia, and acute respiratory syndrome.

23. The method of any one of claims 13 to 22, wherein the subject is afflicted with fever, tiredness, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper respiratory symptoms (e.g. sneezing, runny nose, nasal congestion, cough, sore throat), pneumonia, pneumonia respiratory failure, hepatic and renal insufficiency, acute respiratory distress syndrome (ARDS), and a cytokine imbalance by one or more symptoms of a coronavirus infection relative to an uninfected state.

24. The method of any one of the claims 13 to 23, wherein the method prevents or mitigates the development or progression of the infection.

25. The method of any one of claims 13 to 24, wherein the method prevents or mitigates the development or progression of the infection that requires hospitalization.

26. The method of any one of claims 13 to 25, wherein the method prevents or mitigates the development or progression of the infection that causes death of the subject.

27. The method of any one of claims 13 to 26, wherein the method prevents or mitigates the development or progression of the infection that requires receiving invasive mechanical ventilation.

28. The method of any one of claims 13 to 27, wherein the method prevents or mitigates the development or progression of the infection that requires receiving extracorporeal membrane oxygenation (ECMO).

29. The method of any one of claims 13 to 28, wherein the method prevents or mitigates the development or progression of the infection that requires receiving non-invasive mechanical ventilation.

30. The method of any one of claims 13 to 29, wherein the method prevents or mitigates the development or progression of the infection that requires use of a high flow oxygen device.

31. The method of any one of claims 13 to 30, wherein the method prevents or mitigates the development or progression of the infection that requires receiving supplemental oxygen.

32. The method of any one of claims 13 to 31 , wherein the method prevents or mitigates the development or progression of the infection that requires receiving ongoing medical care.

33. The method of any one of claims 13 to 32, wherein the method causes a decrease in viral load in the subject relative to before treatment.

34. The method of any one of claims 13 to 33, wherein the method reduces or mitigates a likelihood of the subject developing ARDS.

35. The method of any one of claims 13 to 34, wherein the method reduces or mitigates a likelihood of the subject developing lung fibrosis.

36. The method of claim 3 or claim 4, wherein the allergen induces, or is expected to induce, an undesired immune response in a subject.

37. The method of claim 36, wherein the undesired immune response is or comprises a Type 1 hypersensitive immune response.

38. The method of claim 36, wherein the undesired immune response is or comprises allergen-specific antibody production.

39. The method of claim 36, wherein the undesired immune response is or comprises allergen specific CD4+ T cell proliferation and / or activity.

40. The method of claim 36, wherein the undesired immune response is or comprises allergen-specific B cell proliferation and / or activity.41 . The method of any one of the claims 36 to 40, wherein the allergen is or comprises a protein, peptide, lipoproteins, carbohydrate, or are contained or expressed in cells, or fragments or derivatives of the same.

42. The method of any one of the claims 36 to 41 , allergen is or comprises plant allergens (e.g. tree pollen), insect sting allergens (e.g., bee sting allergens), animal allergens (e.g., pet allergens, e.g., animal dander or cat Fel d1 antigen), latex allergens, fungal (including mold and mildew) allergens, cosmetic allergens, drug allergens, food allergens (e.g. mustard), dust, arthropod allergens (e.g. crustacean allergens, insect allergens, arachnid allergen), sulphites, sulphur dioxide, metal allergens (e.g. nickel, cobalt, chromium, zinc), insect venom, viruses, or bacteria.

43. The method of claim 42, wherein the food allergen is selected from milk allergens, egg allergens, nut allergens (e.g., tree nut allergens (e.g., walnuts, cashews, and the like)), fish allergens, crustacean allergens, soy allergens, legume allergens (e.g., peanut allergen), seed allergens and wheat allergens.

44. The method of claim 42, wherein the insect sting allergen is selected from allergens that are or are associated with bee stings, wasp stings, hornet stings, yellow jacket stings and the like.

45. The method of claim 42, wherein the arthropod allergens is selected from insect allergens (e.g., cockroach allergens), crustacean allergens (e.g., lobster, shrimp allergens), and arachnid allergens (e.g., house-dust mites Der p1 antigen) and the like.

46. The method of claim 42, wherein the drug allergen is selected from allergens that are or are associated with antibiotics, NSAIDs, and anaesthetics.

47. The method of claim 42, wherein the pollen allergen, is selected from grass allergens, tree allergens, weed allergens, and flower allergens.

48. The method of claim 42, wherein the metal allergen, is selected from nickel, cobalt, chromium, and zinc.

49. The method of any one of the claims 36 to 48, wherein the allergen is or comprises an asthma antigen, a hay fever antigen, a hives antigen, an eczema antigen, a plant allergen, an insect sting allergen, an insect allergen, an animal allergen, a fungal allergen, a drug allergen, haptens, small chemicals, a pet allergen, a latex allergen, a mold allergen, a cosmetic allergen, or a food allergen.

50. The method of any one of claims 36 to 49, wherein the subject is afflicted with one or more of allergic conditions affecting the nose and eyes (e.g., allergic rhinoconjunctivitis), ears (e.g., allergic otitis media), lungs (e.g., bronchial asthma) as well as for severe insect sting allergy.51 . The method of any one of claims 36 to 50, wherein the subject is afflicted with one or more of fever, tiredness, cough, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper respiratory symptoms (e.g., sneezing, runny nose, nasal congestion, cough, sore throat), itchy and / or painful ears, lower respiratory symptoms, and an immune imbalance relative to a non- allergic state.

52. The method of any one of claims 36 to 51 , wherein the subject has mild or moderate allergies or allergic symptoms.

53. The method of any one of claims 1 to 52, wherein the method prevents or mitigates the development or progression of the viral infection or allergic reaction / response in the subject.

54. The method of claim 53, wherein the method prevents or mitigates the development or progression of the viral infection in a subject including one or more of fever, tiredness, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper respiratory symptoms (e.g. sneezing, runny nose, nasal congestion, cough, sore throat), pneumonia, pneumonia respiratory failure, hepatic and renal insufficiency, lower respiratory symptoms, acute respiratory distress syndrome (ARDS), and a cytokine imbalance.

55. The method of claim 53, wherein the method prevents or mitigates the development or progression of one or more of allergic conditions affecting the nose and eyes (e.g. allergic rhino-conjunctivitis), ears (e.g. allergic otitis media), lungs (e.g. bronchial asthma), skin and / or gastrointestinal tract in a subject, including one or more of fever, tiredness, cough, dry cough, aches and pains, shortness of breath and other breathing difficulties, diarrhea, upper fever, tiredness, cough, dry cough, aches and pains, shortness of breath and other breathing difficulties, asthma, diarrhea, upper respiratory symptoms (e.g. rhinitis, sneezing, runny nose, nasal congestion, cough, sore throat), conjunctivitis, itchy and / or painful ears, lower respiratory symptoms, nausea, vomiting, diarrhea, urticaria, angio-oedema, atopic dermatitis, eczema, eosinophilic disorders (e.g. eosinophilic esophagitis) and an immune imbalance.

56. The method of claim 53, wherein the method prevents or mitigates the development or progression of the infection that requires receiving an additional therapeutic agent.

57. The method of claim 56, wherein the additional therapeutic agent is selected from nirmatrelvir / ritonavir, remdesivir; favipiravir; galidesivir; prezcobix; lopinavir; and / or ritonavir; and / or arbidol lopinavir / ritonavir; and / or ribavirin; and / or IFN-beta;xiyanping; anti-VEGF-A; fingolimod; carrimycin; hydroxychloroquine; darunavir and cobicistat; methylprednisolone; brilacidin; leronlimab; thalidomide, bamlanivimab, casirivimab, and imdevimab.

58. The method of claim 57, wherein the additional therapeutic agent is nirmatrelvir / ritonavir (PAXLOVID).

59. The method of claim 57, wherein the additional therapeutic agent is an antibody directed against an antigen of the virus.

60. The method of claim 59, wherein the additional therapeutic agent is a monoclonal antibody directed against a coronavirus antigen.61 . The method of claim 59, wherein the additional therapeutic agent is a monoclonal antibody directed against an influenza antigen.

62. The method of claim 61 , wherein the additional therapeutic agent is a steroid.

63. The method of claim 62, wherein the steroid is dexamethasone or prednisone.

64. The method of claim 55, wherein the method prevents or mitigates the development or progression of the allergy that requires receiving an additional therapeutic agent (for e.g., an allergy medication).

65. The method of claim 64, wherein the additional therapeutic agent is selected from an over-the-counter (OTC) corticosteroid, a prescription corticosteroid, an over- the-counter (OTC) antihistamine, a prescription antihistamine, over-the-counter (OTC) decongestant, a prescription decongestant, mast-cell stabilizers, leukotriene inhibitors, biological medications and / or emergency epinephrine injections.

66. The method of claims 64 or 65, wherein the additional therapeutic agent is selected from pill, liquids, nasal sprays and / or drops, throat sprays and / or drops, and eye drops.

67. The method of any one of claims 1 to 66, wherein the method restores the immune imbalance and / or restores homeostasis in the subject relative to an untreated or pre-treated state.

68. The method of any one of the claims 1 to 67, wherein the viral infection or allergic response is assayed or assayable in a biological sample from the subject.

69. The method of claim 68, wherein the biological sample is or comprises respiratory fluid, sputum, saliva, or stool.

70. The method of claim 69, wherein the respiratory fluid is from an oropharyngeal (OP) or nasopharyngeal (NP) swab.

71. The method of claim 68 or claim 70, wherein the respiratory fluid is or comprises a nasal secretion.

72. The method of any of the claims 69 to 71 , wherein the respiratory fluid is or comprises saliva.

73. The method of claim 68, wherein the viral infection or allergic reaction / response is assayed or assayable using a measurement of a nucleic acid or protein.

74. The method of claim 73, wherein the viral infection or allergic reaction / response is assayed or assayable using PCR.

75. The method of claim 73, wherein the viral infection is assayed or assayable using a rapid test.

76. The method of claim 73, wherein the allergy or allergic response is assayed or assayable using a patch test.

77. The method of claim 73, wherein the allergy or allergic response is assayed or assayable using a skin prick test.

78. The method of any one of the claims 1 to 77, wherein the GM-CSF has an amino acid sequence of SEQ ID NO: 1 , or a variant of at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98% identity thereto.

79. The method of any one of the claims 1 to 78, wherein the GM-CSF has an amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 3, or a variant of at least about 90%, or at least about 93%, or at least about 95%, or at least about 97%, or at least about 98% identity thereto.

80. The method of any one of the claims 1 to 79, wherein the GM-CSF is one of molgramostim, sargramostim, and regramostim.

81. The method of claim 80, wherein the GM-CSF is sargramostim.

82. The method of any one of the claims 1 to 81 , wherein the GM-CSF is administered at a total dose of about 125 pg, about 150 pg, or about 200 pg, or about 250 pg, or about 300 pg, or about 350 pg.

83. The method of claim 82, wherein the GM-CSF is administered at a total dose of about 250 pg.

84. The method of any one of the claims 1 to 83, wherein the GM-CSF is administered at a dose of about 125 pg, about 150 pg, or about 200 pg, or about 250 pg, or about 300 pg, or about 350 pg, once daily.

85. The method of claim 84, wherein the GM-CSF is sargramostim, administered at a dose of about 250 pg, once daily.

86. The method of any one of the claims 1 to 85, wherein the GM-CSF is sargramostim, administered at a dose of about 250 pg, once daily, for 5 consecutive days.

87. The method of any one of the claims 1 to 86, wherein the GM-CSF is administered by inhalation.

88. The method of any one of the claims 1 to 87, wherein the GM-CSF is administered to the lung.

89. The method of claim 88, wherein the GM-CSF is administered via aerosol or nebulizer.

90. The method of claim 89, wherein the aerosol or nebulizer is selected from liquid nebulization, dry powder dispersion and meter-dose administration.91 . The method of any one of the claims 1 to 90, wherein the method further comprises administering one or more additional therapeutic agents to treat the viral infection or an allergic reaction / response.

92. The method of claim 91 , wherein the additional therapeutic agent to treat the viral infection is selected from nirmatrelvir / ritonavir, remdesivir; favipiravir; galidesivir; prezcobix; lopinavir; and / or ritonavir; and / or arbidol lopinavir / ritonavir; and / or ribavirin; and / or IFN-beta; xiyanping; anti-VEGF-A; fingolimod; carrimycin; hydroxychloroquine; darunavir and cobicistat; methylprednisolone; brilacidin; leronlimab; thalidomide, bamlanivimab, casirivimab, and imdevimab.

93. The method of claim 92, wherein the additional therapeutic agent is nirmatrelvir / ritonavir (PAXLOVID).

94. The method of claim 92, wherein the additional therapeutic agent is an antibody directed against an antigen of the virus.

95. The method of claim 94, wherein the additional therapeutic agent is a monoclonal antibody directed against a coronavirus antigen.

96. The method of claim 94, wherein the additional therapeutic agent is a monoclonal antibody directed against an influenza antigen.

97. The method of claim 94, wherein the additional therapeutic agent is a steroid.

98. The method of claim 97, wherein the steroid is dexamethasone or prednisone.

99. The method of claim 91 , wherein the additional therapeutic agent to treat the allergic reaction / response is selected from an over-the-counter (OTC) corticosteroid, a prescription corticosteroid, an over-the-counter (OTC) antihistamine, a prescription antihistamine, over-the-counter (OTC) decongestant, a prescription decongestant, mast-cell stabilizers, leukotriene inhibitors, biological medications and / or emergency epinephrine injections.

100. The method of any one of claims 1 to 99, wherein the method further comprises the step of monitoring the viral load or allergy symptoms during the course of treatment.

101. The method of claim 100, wherein the monitoring of viral load is assayed or assayable using a measurement of viral nucleic acid or protein.

102. The method of claim 100, wherein the monitoring of viral load is assayed or assayable using PCR.

103. The method of claim 100, wherein the monitoring of viral load is assayed or assayable using a rapid test.

104. The method of claim 100, wherein the monitoring of viral load is assayed or assayable using a lateral flow antigen test.

105. The method of claim 100, wherein an increase in viral load in the subject directs continued administration of GM-CSF.

106. The method of claim 100, wherein a worsening of symptoms in the subject with a viral infection at any timepoint directs toward continued administration of the GM-CSF.

107. The method of claim 100, wherein a decrease in viral load directs towards discontinuation of administration of GM-CSF or a reduction in dosage of GM-CSF.

108. The method of claim 100, wherein a worsening of allergy symptoms in the subject with an allergic reaction at any timepoint directs toward continued administration of the GM-CSF.

109. The method of claim 100, wherein a decrease in an allergic reaction and / or symptoms directs towards discontinuation of administration of GM-CSF or a reduction in dosage of GM-CSF.

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