Use of albumin for modulating immune cells

Abstract

The present invention relates to the use of albumin for modulating immune cells in a subject in need thereof. In particular, the present invention relates to a composition comprising human albumin for use in modulating immune cells in a subject suffering from systemic inflammatory response syndrome.

Description

Technical Field

[0001] This invention relates to the use of albumin in modulating immune cells in subjects in need. In particular, this invention relates to a composition comprising human albumin for modulating immune cells in subjects suffering from systemic inflammatory response syndrome. Background Technology

[0002] The immune system is an aggregate of cells and molecules that work together to protect humans from infectious agents. The immune system also provides humans with a surveillance mechanism to continuously monitor the integrity of cells or tissues. Through this surveillance, changes in major histocompatibility complex proteins or viral proteins on the cell membrane can be identified, and these changes can indicate cancer development or intracellular viral infection. The immune system operates according to two main principles: (1) highly sensitive plasma membranes or intracellular receptors present in immune cells recognize foreign (non-self) substances, which leads to the expression of hundreds of genes that regulate inflammatory responses and immune cell proliferation; (2) eliminate microorganisms through a variety of mechanisms, including non-specific antimicrobial proteins synthesized by the liver (i.e., the complement system, which kills bacteria by directly cleaving or activating phagocytosis by immune cells); the release of intracellular cytotoxic granules by granulocytes, T-cytotoxic lymphocytes (Tc cells), and natural killer (NK) cells, which contain a range of enzymes and perforating proteins (perforin) capable of killing microorganisms, phagocytosis (granulocytes and macrophages), and specific antibodies [B-lymphocytes (B cells)]. Activated Tc and NK cells present Fas-ligands, which interact with Fas-receptors on target cells (cancer or virus-infected cells), leading to cell death caused by apoptosis. Antibodies, complement, and granulocytes defend against most extracellular organisms, while macrophages, Tc cells, and NK cells participate in surveillance processes and eliminate tumor cells and virus-infected cells. Each part of the immune system works in a complementary manner.

[0003] The innate immune system acts as a rapid-response force, deploying a range of non-specific (but highly effective) weapons within minutes to eliminate infectious agents or control the infection contained within. Cells from the innate immune system use a conserved non-specific pattern—recognition receptor (PRR)—that reliably recognize common pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) released during tissue damage. The recognition of PAMPs and DAMPs and the initial immune response is a simple process that occurs within minutes. The innate immune system consists of granulocytes (neutrophils, basophils, and eosinophils), macrophages (histiocytes derived from circulating monocytes), and dendritic cells (specialized cells that present antigens to T, B, and NK cells for an immune response). NK cells, although lymphocytes, are also part of the innate immune system.

[0004] The association of systemic inflammation, immunosuppression, high prevalence of secondary bacterial infections, and multiple organ failure has identified a syndrome common in critically ill patients, including those with acute decompensated cirrhosis, acute liver failure, severe sepsis (including septic shock), acute pancreatitis and other intra-abdominal inflammatory processes, multiple trauma, or severe burns.

[0005] Acute decompensated cirrhosis is defined as the recent onset of ascites, hepatic encephalopathy, gastrointestinal bleeding, or any combination thereof. 1,2 It has two severe forms, including acute-on-chronic liver failure (ACLF). 1,2 And the early stages of ACLF. 3,4 ACLF is defined as a patient with single or multiple organ failure. 1,2 Immune activation was more pronounced in patients with acute decompensated cirrhosis who developed acute-on-chronic liver failure (ACLF) than in those without ACLF (Weiss et al., 2021, Frontiers in Immunol, vol. 12, art. 699563). ACLF is a syndrome characterized by intense systemic inflammation, multiple organ dysfunction / failure, and a very high prevalence of bacterial infections (Moreau et al., 2013, Gastroenterology, 144:1426-37, 1437.e1-9; Arroyo et al., 2020, N Engl J Med, 382:2137-2145). ACLF pre-definition refers to a group of patients with acute decompensated cirrhosis who did not have ACLF at admission but developed ACLF during hospitalization or within 3 months of admission. 3,4 Recent unpublished research suggests two distinct phenotypes of pre-ACLF: "early" pre-ACLF, defined as patients developing ACLF during their initial hospitalization (baseline hospitalization, index hospitalization), and "delayed" pre-ACLF, defined as patients developing ACLF between discharge from their initial hospitalization and the end of a 3-month follow-up period. Studies of patients with ACLF have shown that these patients exhibit strong systemic inflammation (characterized by leukocytosis, ... 1,2 Increased neutrophils 5 Cytokines in the blood 6 Bioactive lipids 7 and C-reactive protein 1,2 Elevated levels indicate immunosuppression (response to a reduced response of a specific subset of monocytes to bacterial products); 8-10 Neutrophils have a reduced ability to kill microorganisms; 5 And lymphopenia involving T cells, B cells, and NK cells.5 (Indications), and the high incidence of secondary infections, 11,12 These are all fatal complications. 11 Patients hospitalized for the "early" pre-ACLF stage also exhibited severe systemic inflammation, while those with the "slowly progressive" pre-ACLF stage showed moderate inflammation. To date, there are no therapies targeting the immune cellular dysregulation in these patients.

[0006] Albumin is commonly used for patients with sclerosis, including those treated with large-volume biopsies. 13 and those suffering from spontaneous bacterial peritonitis (SBP) 14 Or hepatorenal syndrome-acute kidney injury (HRS-AKI) 15 The effect of albumin is usually attributed to plasma volume expansion in patients. 11 However, recent studies have shown that albumin may affect the function of specific immune cells. 16,17 These findings may explain the lower incidence of SBP and other infections reported in patients receiving long-term albumin administration compared to patients receiving standard drug therapy. 18

[0007] In addition, although there is no established treatment for immunosuppression to date, recent studies have shown that intravenous albumin, in addition to its plasma dilator properties, can reduce the severity of systemic inflammation (Fernández et al., 2018, Gut, 67:1870-1880; Casulleras et al., 2020, Sci Transl Med, 12(566):eaax5135). Summary of the Invention

[0008] The effects of intravenous albumin on characteristic lymphopenia and defective neutrophil antimicrobial function in patients with acute-on-chronic liver failure (ACLF) are unclear.

[0009] This invention is based on the surprising discovery that treatment with human albumin affects different subsets of immune cells (specifically neutrophils, CD4+ T cells, dendritic cells, and / or monocytes) in subjects. Therefore, compositions containing human albumin have beneficial utility in modulating immune cells (particularly neutrophils, CD4+ T cells, dendritic cells, and / or monocytes) in subjects in need. This invention is particularly useful, for example, in the prevention of microbial infections, in subjects with impaired immune cells (particularly neutrophils, CD4+ T cells, dendritic cells, and / or monocytes) and / or in subjects who could benefit from increased immune cell function (particularly neutrophil function, CD4+ T cell function, dendritic cell function, and / or monocyte function).

[0010] As described in the examples included herein, the inventors studied forty-nine patients admitted for severe acute decompensated cirrhosis (without ACLF) and performed whole blood RNA sequencing (RNA-seq) at admission and 15 days after the onset of ACLF. These patients were selected because they followed a stable systemic inflammatory process. Thirty patients received albumin during the progression to ACLF, but the other 19 did not. Furthermore, in other patients with acute decompensated cirrhosis, the inventors performed single-cell RNA-seq (scRNA-seq) on peripheral blood mononuclear cells (PBMCs) exposed to albumin or a vehicle for 2 hours in vitro and assessed the antimicrobial activity of neutrophils exposed to albumin in vitro.

[0011] Whole blood RNA-seq data analysis showed that patients who received albumin exhibited specific upregulation of signatures associated with B cells, plasma cells, and immunoglobulins; CD4 T cells; myeloid cells; mismatch repair, cell cycle, and mitosis; and transcription factors such as c-Myc and E2F family members. Analysis of patient PBMCs exposed in vitro to albumin using scRNA-seq revealed increased signatures associated with B cells, myeloid cells, and CD4 T cells. Furthermore, the inventors demonstrated that neutrophils exposed in vitro to albumin exhibited increased degranulation and enhanced phagocytosis.

[0012] Therefore, it is advantageous that the inventors have demonstrated that, in patients with severe acute decompensated cirrhosis, albumin promotes the expansion of B cell compartments and CD4 T cell compartments, acts on monocytes and myeloid cells, and restores the antimicrobial function of neutrophils to normal.

[0013] Therefore, this article provides a composition comprising human albumin for modulating immune cells in subjects suffering from systemic inflammatory response syndrome, wherein: (a) The composition comprising human albumin is used to modulate neutrophil function, wherein the human albumin is administered to a subject at a dose sufficient to increase the antimicrobial function of neutrophils; and / or (b) The composition comprising human albumin is used to modulate CD4+ T cells, wherein the human albumin is administered to a patient at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells; and / or (c) The composition comprising human albumin is used to modulate dendritic cells, wherein the human albumin is administered to a subject at a dose sufficient to increase the level of plasma cell-like dendritic cells; and / or (d) The composition comprising human albumin is used to regulate monocytes, wherein the human albumin is administered to a subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

[0014] Appropriately, an increase in the antimicrobial function of neutrophils may be due to an increase in neutrophil degranulation and / or neutrophil phagocytosis.

[0015] Human albumin may be administered to a subject in a dose sufficient to upregulate the expression of neutrophil genes selected from the following: CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1, or combinations thereof.

[0016] Appropriately, systemic inflammatory response syndrome may be associated with a selection of diseases including: decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

[0017] Appropriately, subjects may be patients with sepsis or liver disease.

[0018] Appropriately, subjects may have liver conditions selected from the following: decompensated cirrhosis, pre-acute-on-chronic liver failure, and acute-on-chronic liver failure.

[0019] A composition comprising human albumin is also provided for treating defective immune cell function in subjects in need, wherein the defective immune cell function is selected from: (a) Defective neutrophil function; (b) Defective central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function; (c) Defective plasmacytoid dendritic cell function; and (d) Defective intermediate HAVCR2+ monocyte function.

[0020] Appropriately, the treatment may increase the antimicrobial function of neutrophils in the subject, optionally wherein the treatment may increase neutrophil degranulation and / or neutrophil phagocytosis.

[0021] Human albumin may be administered to a subject in a dose sufficient to upregulate the expression of neutrophil genes selected from the following: CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1, or combinations thereof.

[0022] Appropriately, the defective immune cell function may be associated with diseases selected from: systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

[0023] Where appropriate, human albumin may be administered via a route of administration selected from the following: intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

[0024] Human albumin can be used as part of a multi-dose regimen to administer the drug at multiple intervals.

[0025] Suitable, a multiple dosing regimen may comprise multiple fractional doses administered at equal intervals of about 1 day to about 30 days; or the multiple dosing regimen may comprise multiple fractional doses administered at unequal intervals of about 1 day to about 30 days; or the multiple dosing regimen may comprise two or more doses up to the total cumulative dose.

[0026] Appropriately, human albumin may be administered as part of a multiple-dose regimen at a single dose (single dose) of about 5 g / interval (g / interval, g / time period) to about 500 g / interval; or human albumin may be administered as part of a multiple-dose regimen at a single dose of about 20 g / interval to about 200 g / interval.

[0027] Appropriately, the dosing interval can be every 15 days or less.

[0028] Appropriately, human albumin may be albumin derived from human plasma or recombinant human albumin.

[0029] Appropriately, the concentration of human albumin may be between 4% and 25% (w / v); or the concentration of human albumin may be about 20% (w / v).

[0030] As described herein, human albumin affects different subsets of immune cells (particularly neutrophils, CD4+ T cells, dendritic cells, and / or monocytes) in subjects. Therefore, it is a therapeutic agent in the compositions described herein. Therefore, the term "human albumin" is used interchangeably with the term "composition containing human albumin" throughout.

[0031] Throughout the description and claims of this specification, the words “comprising” and “containing” and their variations mean “including but not limited to”, and they are not intended (and will not) exclude other parts, additives, components, integers or steps.

[0032] Throughout the description and claims of this specification, the singular encompasses the plural unless the context requires otherwise. In particular, when the indefinite article is used, the specification should be understood to include both the plural and the singular unless the context requires otherwise.

[0033] The features, integers, properties, compounds, chemical parts or groups described in connection with a particular aspect, embodiment or example of the invention should be understood to be applicable to any other aspect, embodiment or example described herein, unless incompatible therewith.

[0034] The various aspects of the present invention will now be described in further detail. Attached Figure Description

[0035] Embodiments of the present invention are further described below with reference to the accompanying drawings, wherein: Figure 1. Whole blood gene signatures associated with albumin treatment. (A) Euler plot showing differentially expressed genes (DEGs) between time 2 (T2) and time 1 (T1) in the albumin and non-albumin groups. DEG is defined by an absolute fold change (FC) greater than 1.5 and P < 0.05. (B) Volcano plot showing the relationship between the differential expression effect size (log2 FC) and significance (-log10 P) between T2 and T1 for the albumin group (top) and the non-albumin group (bottom). In both volcano plots, gray dots indicate genes for which there is no significant difference in expression between T2 and T1 (absolute FC less than 1.5 and P > 0.05, i.e., -log10 P < 1.3). Gray dots indicate DEGs, which are genes that are upregulated or downregulated. (C) Differential expression effect size (log2 FC) between T2 and T1 in the albumin group compared to the corresponding differential effect size in the non-albumin group. This section only shows genes included in the GeneOntology gene set “GOCC_Immunoglobulin Complex”. DEGs with a concordant sign are considered shared. (D) Violin plot of RNA-seq deduced features of plasmablasts, NK cells, and T cells at T1 and T2 in the albumin- and non-albumin groups, determined using SingleRR software. Baseline values ​​for healthy subjects are also shown. White diamond symbols indicate the median. P-values ​​are derived from the Kruskal–Wallis test and subsequent Mann–Whitney U test. (E) Heatmap of 32 differentially expressed (DE) blood transcriptional modules (BTMs) between T2 and T1, which are specific to either the albumin- or non-albumin-group. Thirty-one DE BTMs are specific to the albumin-group, while only one DE BTM (“Endoplasmic Reticulum (M37.2)”) is specific to the non-albumin-group. Hierarchical clustering of BTM was performed based on the QuSAGE activity fraction obtained from the albumin group. An asterisk indicates P < 0.05. Gray values ​​represent QuSAGE activity fractions (-0.4 to 0.4).

[0036] Figure 2. Single-cell RNA sequencing identifies specific immune cell changes in isolated peripheral blood mononuclear cells (PBMCs) exposed to albumin. This figure was designed using results obtained from PBMCs from patients with acute decompensated cirrhosis. (A) Uniform manifold approximation and projection (UMAP) of B lymphocytes exposed to albumin and mediators in 1946 patients, colored by cell type. (B) Overlay plot of UMAP of albumin and mediators exposed to B lymphocytes. (C) Box plot of B lymphocyte population abundance that changed significantly after albumin exposure. (D) UMAP of myeloid cells exposed to albumin and mediators in 21819 patients, indicated by cell population. (E) Overlay plot of myeloid cell UMAP of albumin and mediators exposed to mediators. (F) Box plot of myeloid cell type (plasma cell-like dendritic cells; pDC) abundance that changed significantly after albumin exposure. (G) UMAP of CD4 T cells exposed to albumin and mediators in 12692 patients, colored by cell type. (H) Overlay plot of albumin and mediator exposure on CD4 T cells UMAP. (I) Box plot of significant changes in CD4 T cell abundance after albumin exposure. (J) Left panel, representative density plot showing the characteristic fractions of the mitotic cell cycle in BTMP-stimulated CD4 T cells (M4.11) on CD4 T cell UMAP under mediator and albumin conditions. Right panel, comparison of log(characteristic fractions) of BTM 4.11 between albumin and mediator in CD4 T cell compartments. Statistical analysis of BTM characteristics was performed using the Wilcoxon signed-rank test; p-values ​​indicated significance. Significance of comparisons of abundance across all cell types between albumin and mediator was tested using the paired Wilcoxon signed-rank test, with adjusted p-values ​​indicating significance.

[0037] Figure 3 Effects of albumin on host defense function of neutrophils from patients with acute decompensated cirrhosis. Figures A and B are designed using functional assays from freshly isolated peripheral neutrophils from 6 patients with acute decompensated cirrhosis and 5 age-matched healthy subjects. (A) Neutrophils from patients with acute decompensated cirrhosis and healthy subjects were incubated with cell culture medium (medium), human serum albumin (15 mg / mL), or recombinant human albumin (15 mg / mL) in a 5% CO2 incubator at 37°C for 2 h. Neutrophil degranulation was assessed by measuring MPO enzyme activity in the cell supernatant. (B) Phagocytic capacity was assessed by incubating neutrophils from patients with acute decompensated cirrhosis and healthy subjects with FITC-coupled yeast polysaccharide bioparticles alone or in the presence of human serum albumin and recombinant albumin (15 mg / mL) for 60 min and comparing them with a medium control.

[0038] The patents, scientific and technical documents cited herein establish the knowledge available to a person skilled in the art at the time of filing the application. All disclosures of granted patents, published and pending patent applications, and other publications cited herein are incorporated herein by reference as if each were specifically and individually indicated to be incorporated by reference. In any event of inconsistency, this disclosure shall prevail.

[0039] The various aspects of the present invention are described in further detail below. Detailed Implementation

[0040] This invention is based on the surprising finding that treatment with human albumin affects different subsets of immune cells in the subject’s body (specifically neutrophils, CD4+ T cells, dendritic cells and / or monocytes).

[0041] Following albumin treatment, the immune cell gene signatures of patients with acute decompensated cirrhosis (pre-ACLF who developed ACLF) (“albumin group”) were compared with those of equivalent patients who did not receive albumin (“no albumin group”). Significant numerical similarities were found between the transcriptional signatures of pre-ACLF and ACLF patients. Furthermore, differential expression of several gene modules was observed in the albumin group. Specifically, several genes acting as markers of activated low-density neutrophils were upregulated in patients treated with albumin. Additionally, a significant increase in the abundance of intermediate monocyte HAVCR2+ and plasmacytoid DCs was observed in the albumin group. Figure 2E and 2F Finally, albumin was shown to alter the profile of CD4+ T cell compartments. Figure 2H The abundance of activated memory CD4+ T cells decreased significantly, while the abundance of central memory ITGB1+ CD4+ T cells increased. Figure 2I ).

[0042] Patients with acute decompensated cirrhosis have a very high prevalence of bacterial infection upon admission. A significant proportion of patients who are not infected upon admission develop bacterial infections during hospitalization due to severely impaired antimicrobial defense mechanisms. The results of this study indicate that albumin can salvage deficient neutrophil antimicrobial function and depleted lymphocyte compartments in the most severe forms of acute decompensated cirrhosis.

[0043] Regulation of neutrophils

[0044] The inventors have demonstrated in this paper that treatment with human albumin can specifically upregulate nine major neutrophil genes in subjects. CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, MCEMP1 () Figure 1B Furthermore, this study demonstrates that neutrophils exposed to human albumin in vitro surprisingly exhibit increased antimicrobial activity, including increased degranulation and enhanced phagocytosis. Figure 3 The data presented in this article indicate that human albumin can modulate neutrophil function in subjects (particularly increasing the antimicrobial function of neutrophils).

[0045] Therefore, this document provides a composition comprising human albumin for modulating neutrophil function in a subject, wherein human albumin is administered to / is administered to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis). Such compositions can be used to prevent infection and / or inhibit pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in a subject. Therefore, this document also provides a composition comprising human albumin for preventing infection and / or inhibiting pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in a subject, wherein human albumin is administered to / is administered to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis).

[0046] In one example, this article provides a composition comprising human albumin for preventing microbial infection and / or inhibiting pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in a subject, wherein human albumin is administered to / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., wherein the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis).

[0047] Examples of suitable microbial infections include bacterial infections such as spontaneous bacterial peritonitis (SBP). SBP is one of the most common infections in patients with cirrhosis. The most frequently involved pathogens in SBP are Enterobacteriaceae and non-enterococcal streptococci, with enterococci being increasingly common in nosocomial episodes. Among other infections, the most common are urinary tract infections (UTI), pneumonia, primary / spontaneous bloodstream infections (BSI), and skin and soft tissue infections. The most frequently involved pathogens in UTI are Enterobacteriaceae and enterococci, while the most frequently involved pathogens in primary BSI are Enterobacteriaceae and staphylococci. Other less common infections include cholangitis, endocarditis, catheter-related bloodstream infections, Clostridium difficile enterocolitis, and secondary peritonitis. See Fernandez et al., Management of bacterial and fungal infections in cirrhosis: The MDRO challenge, Journal of Hepatology, Volume 75, Supplement 1, July 2021, pp. S101-S117. These appropriate examples of microbial infections apply to all relevant aspects described herein.

[0048] The compositions provided herein are particularly useful when administered to subjects suffering from systemic inflammatory response syndrome (SIRS). Subjects with SIRS typically have deficient neutrophil antimicrobial function. Advantageously, the compositions provided herein can be used to increase neutrophil antimicrobial function in these subjects (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis), and therefore can be specifically used to prevent infection and / or inhibit pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects with SIRS. Therefore, this document provides a composition comprising human albumin for modulating neutrophil function in subjects with SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis).

[0049] Therefore, the compositions provided herein can be used to treat SIRS in subjects by modulating neutrophil function. This document also provides a composition comprising human albumin for preventing infection and / or inhibiting pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects with SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., wherein the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis). Therefore, the compositions provided herein can be used to treat SIRS in subjects by preventing infection and / or inhibiting pathogen-associated molecular pattern (PAMP)-induced systemic inflammation.

[0050] Examples of subjects with SIRS are described elsewhere in this article. For example, SIRS may be associated with diseases selected from the following: decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. In other words, subjects with SIRS may also have diseases selected from the groups listed above.

[0051] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or a subject with ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0052] This document also provides a composition comprising human albumin for modulating neutrophil function in a subject with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein human albumin is administered / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., wherein the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis). Therefore, the composition provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in a subject by modulating neutrophil function. Furthermore, this article provides a composition comprising human albumin for the prevention of infection and / or inhibition of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in patients with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein human albumin is administered / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis). Therefore, the composition provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by preventing infection and / or inhibiting their pathogen-associated molecular pattern (PAMP)-induced systemic inflammation.

[0053] The compositions provided herein are also useful when administered to subjects with defective neutrophil function (e.g., defective neutrophil antimicrobial function, where the defective neutrophil antimicrobial function may be defective neutrophil degranulation and / or defective neutrophil phagocytosis)). Advantageously, the compositions provided herein can be used to increase neutrophil antimicrobial function in these subjects (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis), and can therefore be specifically used to prevent infection and / or inhibit pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects with defective neutrophil function. Therefore, this document provides a composition comprising human albumin for treating defective neutrophil function in subjects in need. In this example, human albumin is administered / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis). This article also provides a composition comprising human albumin for the prevention of infection and / or inhibition of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects with defective neutrophil function. In this example, human albumin is administered / given to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., where the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis).

[0054] Those skilled in the art can readily identify examples of subjects with defective neutrophil function. Defects in neutrophil function may manifest as defects in various biological processes related to neutrophil bactericidal activity, including defects in endothelial blockade or adhesion, defects in oxidative burst, defects in extracellular bactericidal mechanisms such as neutrophil extracellular traps or NETS release, defects in neutrophil degranulation, and / or defects in neutrophil phagocytosis.

[0055] For example, deficient neutrophil function may be associated with diseases selected from the following: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, deficient neutrophil function may be present in subjects with diseases selected from the following: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, the compositions comprising human albumin described herein can be used to treat defective neutrophil function selected from the following diseases: SIRS, decompensated cirrhosis, acute liver failure, pre-acute liver failure, chronic-on-acute liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer (in other words, the compositions can be used to treat defective neutrophil function in the aforementioned diseases). Therefore, this article also provides a composition comprising human albumin for treating diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer, wherein the composition is used to modulate neutrophil function (e.g., thereby treating defective neutrophil function).

[0056] In one example, a subject with defective neutrophil function is a subject with decompensated cirrhosis. For example, a subject with defective neutrophil function could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0057] In one example, the composition provided herein comprises human albumin, wherein the human albumin is administered to a subject in a dose sufficient to upregulate the expression of neutrophil genes selected from: CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1, or combinations thereof. This example applies to all suitable aspects provided herein, including when the composition comprising human albumin is used to modulate the immune cells of a subject (e.g., a subject with SIRS and / or a subject with decompensated cirrhosis), or when the composition comprising human albumin is used to treat defective neutrophil function in a subject in need (where applicable).

[0058] CD177 (NCBI gene ID: 57126; UniProtKB / Swiss-Prot: Q8N6Q3) encodes a glycosyl-phosphatidylinositol (GPI)-linked cell surface glycoprotein that plays a role in neutrophil activation. This protein can bind to platelet endothelial cell adhesion molecule-1 and functions in neutrophil migration. Mutations in this gene are associated with myeloproliferative disorders. Overexpression of this gene has been found in patients with polycythemia vera. Autoantibodies against this protein may cause pulmonary transfusion reactions, and it may be involved in Wegener's granulomatosis.

[0059] Oligosin 4 (OLFM4 - NCBI gene ID: 10562; UniProtKB / Swiss-Prot: Q6UX06) is a glycoprotein primarily expressed in myeloid cells and gastrointestinal tissues. OLFM4 is stored in specific granules of human neutrophils, defining a subset of neutrophils ranging from 5% to 40% that are OLFM4 positive. OLFM4 is thought to inhibit cathepsin C, a cysteine ​​protease crucial for the activation of serine proteases.

[0060] The proteoglycan 2 (PRG2) gene (NCBI gene ID: 5553; UniProtKB / Swiss-Prot: P13727, also known as the pro eosinophil major basic protein) encodes a major component of the eosinophil granule crystal core. High levels of the proform of PRG2 protein are also found in placental and pregnancy serum, where it exists in complexes with various other proteins, including pregnancy-associated plasma protein A (PAPPA), angiotensinogen (AGT), and C3dg. PRG2 protein may participate in antiparasitic defense mechanisms as a cytotoxin and helmintoxin, and is involved in immune hypersensitivity responses. PRG2 protein contains a peptide that exhibits potent antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria, and fungi. It is directly associated with epithelial cell damage, shedding, and bronchospasm in allergic diseases.

[0061] Myeloperoxidase (MPO - NCBI gene ID: 4353; UniProtKB / Swiss-Prot: P05164) is a heme protein synthesized during myeloid differentiation and is a major component of azurophilic granules in neutrophils. MPO is produced as a single-chain precursor, which is subsequently cleaved into light and heavy chains. Mature myeloperoxidase is a tetramer composed of two light chains and two heavy chains. MPO produces hypohalite, which is crucial for the bactericidal activity of neutrophils.

[0062] Bactericidal permeability-increasing protein (BPI - NCBI gene ID: 671; UniProtKB / Swiss-Prot: P17213) encodes a lipopolysaccharide-binding protein. It is associated with human neutrophil granules and exhibits antimicrobial activity against Gram-negative bacteria. BPI is a potent neutrophil antimicrobial protein with bactericidal and LPS-neutralizing activity, and may also stimulate neutrophil phagocytosis of Gram-negative bacteria by promoting complement activation.

[0063] Resistin (RETN - NCBI gene ID: 56729; UniProtKB / Swiss-Prot: Q9HD89) belongs to the mouse resistin-like gene family. This family is characterized by C-terminal stretches (segments) of 10 cysteine ​​residues with equal spacing. RETN proteins exhibit antimicrobial activity in the skin, showing antimicrobial activity against both Gram-positive and Gram-negative bacteria. RETN promotes pro-inflammatory activation of neutrophils and the formation of extracellular traps in neutrophils.

[0064] Lipocalcin 2 (LCN2 - NCBI gene ID: 3934; UniProtKB / Swiss-Prot: P80188) encodes a protein belonging to the lipocalcin family. Members of this family transport small hydrophobic molecules, such as lipids, steroid hormones, and retinoids. LCN2 is a neutrophil gelatinase-associated lipocalcin and plays a role in innate immunity by restricting bacterial growth through the isolation of iron-containing siderophores. The presence of LCN2 protein in blood and urine is an early biomarker of acute kidney injury. LCN2 is thought to be involved in various cellular processes, including maintaining skin homeostasis and inhibiting invasiveness and metastasis. Mice lacking this gene are more susceptible to bacterial infections than wild-type mice.

[0065] CEA-CAM8 (NCBI gene ID: 1088; UniProtKB / Swiss-Prot: P31997) is expressed exclusively on granulocytes and encodes a cell surface glycoprotein that functions in a calcium-independent manner in cell adhesion. CEACAM8 mediates heterophilic cell adhesion (heterogeneous cell adhesion) with other carcinoembryonic antigen-associated cell adhesion molecules, such as CEACAM6. Furthermore, heterophilic interactions with CEACAM8 (heterogeneous interactions) occur in activated neutrophils.

[0066] Mast cell-expressed membrane protein 1 (MCEMP1 - NCBI gene ID: 199675; UniProtKB / Swiss-Prot: Q8IX19) is a type II transmembrane protein primarily expressed in myeloid immune cells, such as resident mast cells and alveolar macrophages. MCEMP1 is one of the most important inducible genes in many inflammatory diseases, such as asthma, idiopathic pulmonary fibrosis, cancer, sepsis, and stroke.

[0067] As used herein, "regulating neutrophil function" refers to controlling neutrophil function. Neutrophil function can be controlled by increasing or decreasing neutrophil activity (e.g., by stimulating existing neutrophils or by increasing the total number of neutrophils present). In the context of this invention, it is desirable to regulate neutrophil function by increasing neutrophil antimicrobial function (also referred to herein as neutrophil antimicrobial activity). Increased neutrophil antimicrobial function may also be referred to as enhanced neutrophil antimicrobial function.

[0068] Neutrophils are known to perform a variety of antimicrobial functions in vivo. Neutrophil antimicrobial functions include neutrophil degranulation and neutrophil phagocytosis. The increased neutrophil antimicrobial function used in this article can refer to an increase in neutrophil degranulation and / or neutrophil phagocytosis. In some examples, the increased neutrophil antimicrobial function refers to an increase in a combination of these features, namely, an increase in both neutrophil degranulation and neutrophil phagocytosis.

[0069] Methods for determining whether neutrophil antimicrobial function is increased are well known. Examples of such methods are provided in the Examples section below.

[0070] During the host defense response, neutrophils release myeloperoxidase (MPO), an essential antimicrobial protein primarily located in azurophilic or primary neutrophil granules. Therefore, the increase in neutrophil degranulation can be quantified by measuring MPO activity. This increase in neutrophil degranulation may also be referred to herein as enhanced neutrophil degranulation.

[0071] Phagocytosis can be assessed by determining the uptake of fluorescently labeled yeast glycan particles by neutrophils. This method can be used to determine whether neutrophil phagocytosis is increased. An increase in neutrophil phagocytosis may also be referred to herein as enhanced neutrophil phagocytosis.

[0072] Relevant methods are also provided for modulating neutrophil function (and / or preventing infection and / or inhibiting pathogen-associated molecular pattern (PAMP)-induced systemic inflammation) in subjects, wherein a composition comprising human albumin is administered to the subject at a dose sufficient to increase neutrophil antimicrobial function (e.g., wherein the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis). Relevant methods are also provided for treating defective neutrophil function in subjects in need.

[0073] Regulation of CD4+ T cells

[0074] The inventors have demonstrated in this paper that albumin treatment specifically upregulates the mitotic cell cycle in the blood transcription module (BTM) of subjects, specifically in stimulated CD4 T cells (M4.11). Figure 1E This indicates transcriptional activation in CD4+ T cells. Furthermore, this study demonstrates that albumin administration alters the profile of CD4+ T cell compartments in subjects (…). Figure 2H The abundance of activated memory CD4+ T cells decreased significantly while the abundance of central memory ITGB1+ CD4+ T cells increased. Figure 2IThe data presented in this article indicate that albumin can modulate the CD4+ T cell population in subjects (specifically by increasing the level of central memory ITGB1+ CD4+ T cells and decreasing the level of activated CD4+ memory T cells) and alter CD4+ T cell transcription, thereby altering CD4+ T cell function.

[0075] Therefore, this document provides a composition comprising human albumin for modulating CD4+ T cells in a subject, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells. Such compositions can be used to modulate CD4+ T cell function in a subject. Central memory ITGB1+ CD4+ T cells have recently been shown to possess cytotoxic properties (JI 2021,207: 2966-2975), indicating that these cells are involved in host resistance to infection. Therefore, the compositions provided herein can be used to prevent infection in a subject. Therefore, this document also provides a composition comprising human albumin for preventing infection in a subject, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells.

[0076] The compositions provided herein are particularly useful when administered to subjects suffering from systemic inflammatory response syndrome (SIRS). Subjects with SIRS may have defective CD4+ T cell function (particularly insufficient levels of central memory ITGB1+ CD4+ T cells and / or excessive levels of activated CD4+ memory T cells). Advantageously, the compositions provided herein can be used to increase the levels of central memory ITGB1+ CD4+ T cells and / or decrease the levels of activated CD4+ memory T cells in these subjects, and therefore can be specifically used to prevent infection in subjects suffering from SIRS. Therefore, this article provides a composition comprising human albumin for modulating CD4+ T cells (e.g., modulating CD4+ T cell function) in subjects suffering from SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase the levels of central memory ITGB1+ CD4+ T cells and / or decrease the levels of activated CD4+ memory T cells. Therefore, the compositions provided herein can be used to treat SIRS in subjects by modulating their CD4+ T cells (e.g., modulating CD4+ T cell function). This article also provides a composition comprising human albumin for the prevention of infection in a subject suffering from SIRS, wherein the human albumin is administered / given to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells. Therefore, the composition provided herein can be used to treat SIRS in a subject by preventing infection.

[0077] Elsewhere in this article, examples of subjects with SIRS are described. These examples are equally applicable to this aspect.

[0078] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or a subject with ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0079] This article also provides a composition comprising human albumin for modulating CD4+ T cells (e.g., modulating CD4+ T cell function) in a subject with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is administered / given to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells. Therefore, the composition provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in a subject by modulating CD4+ T cells (e.g., modulating CD4+ T cell function). Furthermore, this article provides a composition comprising human albumin for the prevention of infection in subjects with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is administered / given to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells. Therefore, the composition provided herein can be used for the treatment of decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by preventing infection.

[0080] The compositions provided herein are also useful when administered to subjects with defective CD4+ T cell function (e.g., defective central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function). Advantageously, the compositions provided herein can be used to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells in these subjects, and thus can be specifically used for infection prevention in subjects with defective CD4+ T cell function. Therefore, this document provides a composition comprising human albumin for treating defective CD4+ T cell function in subjects in need. In this example, human albumin is administered / given to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells. This document also provides a composition comprising human albumin for infection prevention in subjects with defective CD4+ T cell function. In this example, human albumin was administered to / done to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells.

[0081] Those skilled in the art can readily identify examples of subjects with defective CD4+ T cell function. For instance, defective CD4+ T cell function may be associated with diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, defective CD4+ T cell function may be present in subjects with diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, the compositions containing human albumin described herein can be used to treat defective CD4+ T cell function in a range of diseases including: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer (in other words, the compositions can be used to treat defective CD4+ T cell function in the aforementioned diseases).

[0082] Therefore, this article also provides a composition comprising human albumin for the treatment of diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma syndrome, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer, wherein the composition is used to modulate CD4+ T cell function (e.g., thereby treating defective CD4+ T cell function).

[0083] In one example, a subject with defective CD4+ T cell function is a subject with decompensated cirrhosis. For example, a subject with defective CD4+ T cell function could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0084] As used herein, “regulating CD4+ T cells” refers to controlling CD4+ T cells. This encompasses, for example, controlling the number of CD4+ T cells, controlling the CD4+ T cell population, and controlling CD4+ T cell function. CD4+ T cell function can be controlled by increasing or decreasing CD4+ T cell activity, for example, by stimulating a desired subset of CD4+ T cells (to increase the desired T cell activity of these cells), and / or by increasing the (relative) abundance of desired cells (to increase the number of cells with the desired T cell activity). Similarly, CD4+ T cell function can be controlled by decreasing CD4+ T cell activity, for example, by inactivating an unwanted subset of CD4+ T cells (to decrease the desired T cell activity of these cells), and / or by decreasing the (relative) abundance of unwanted cells (to decrease the number of cells with the unwanted T cell activity). In the context of this invention, it is desirable to regulate CD4+ T cell function by increasing the level of central memory ITGB1+ CD4+ T cells and / or decreasing the level of activated CD4+ memory T cells.

[0085] As used herein, the term "level" refers to the quantity of a specified cell type (e.g., central memory ITGB1+ CD4+ T cells or activated CD4+ memory T cells). This level can be the absolute quantity of cells and / or the relative quantity of the specified cells. The relative quantity of the specified cells can be relative to other cells in the subject's body or from a sample from the subject. These "other cells" can be, for example, all white blood cells, or specific cell subsets, such as CD4+ T cells. The relative quantity can be, for example, a percentage, fraction, or ratio. In the context of relative quantity, it should be understood that the relative quantity of the specified cells can be changed (e.g., increased or decreased) while the absolute quantity of the specified cells may or may not be changed. Similarly, the absolute quantity of the specified cells can be changed (increased or decreased) while the relative quantity may or may not be changed.

[0086] Methods for determining whether the level of central memory ITGB1+ CD4+ T cells is increased and / or whether the number of activated CD4+ memory T cells is decreased are well known. Examples of such methods are provided in the Examples section below.

[0087] Also provided are corresponding methods for modulating CD4+ T cell function (and / or preventing infection) in subjects, wherein a composition comprising human albumin is administered to the subject at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells. Also provided are corresponding methods for treating defective CD4+ T cell function in subjects in need.

[0088] Regulation of dendritic cells

[0089] The inventors have demonstrated in this paper that albumin treatment specifically increases the abundance of plasmacytoid dendritic cells in subjects. Figure 1E and 2D -2F). The data presented in this article indicate that albumin can modulate dendritic cells (particularly by increasing the level of plasmacytoid dendritic cells) and alter transcription in dendritic cells, thereby altering dendritic cell function.

[0090] Therefore, this document provides a composition comprising human albumin for modulating dendritic cells (DCs) in a subject, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells (pDCs). Such compositions can be used to modulate dendritic cell function in a subject. Plasmacytoid DCs are DCs specifically involved in interferon-mediated antiviral responses. Therefore, the compositions provided herein can be used to prevent viral infection in a subject. Therefore, this document also provides a composition comprising human albumin for preventing viral infection in a subject, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells.

[0091] The compositions provided herein are particularly useful when administered to subjects suffering from systemic inflammatory response syndrome (SIRS). Subjects with SIRS may have defective dendritic cell function (particularly insufficient levels of plasmacytoid dendritic cells). Advantageously, the compositions provided herein can be used to increase the levels of plasmacytoid dendritic cells in these subjects and can therefore be specifically used to prevent viral infections in subjects suffering from SIRS. Therefore, this article provides a composition comprising human albumin for modulating dendritic cells (e.g., modulating dendritic cell function) in subjects suffering from SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase the levels of plasmacytoid dendritic cells. Thus, the compositions provided herein can be used to treat SIRS in subjects by modulating dendritic cells (e.g., modulating dendritic cell function). This article also provides a composition comprising human albumin for preventing viral infections in subjects suffering from SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase the levels of plasmacytoid dendritic cells. Thus, the compositions provided herein can be used to treat SIRS in subjects by preventing viral infections.

[0092] Elsewhere in this article, examples of subjects with SIRS are described. These examples are equally applicable to this aspect.

[0093] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or a subject with ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0094] This document also provides a composition comprising human albumin for modulating dendritic cells (e.g., modulating dendritic cell function) in a subject with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells. Therefore, the composition provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in a subject by modulating dendritic cells (e.g., modulating dendritic cell function). Furthermore, this document also provides a composition comprising human albumin for preventing viral infection in a subject with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells. Therefore, the compositions provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by preventing viral infection.

[0095] The compositions provided herein are also useful when administered to subjects with defective dendritic cell function (e.g., defective plasmacytoid dendritic cell function). Advantageously, the compositions provided herein can be used to increase the level of plasmacytoid dendritic cells in these subjects and can therefore be specifically used to prevent viral infections in subjects with defective dendritic cell function. Therefore, this document provides a composition comprising human albumin for treating defective dendritic cell function in subjects in need. In this example, human albumin is administered / given to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells. This document also provides a composition comprising human albumin for preventing viral infections in subjects with defective dendritic cell function. In this example, human albumin is administered / given to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells.

[0096] Those skilled in the art can readily identify examples of subjects with defective dendritic cell function. For instance, defective dendritic cell function may be associated with diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, defective dendritic cell function may be present in subjects with diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, the compositions comprising human albumin described herein can be used to treat defective dendritic cell function in a selection of diseases including: SIRS, decompensated cirrhosis, acute liver failure, pre-acute chronic liver failure, chronic acute liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer (in other words, the compositions can be used to treat defective dendritic cell function in the diseases mentioned above).

[0097] Therefore, this article also provides a composition comprising human albumin for the treatment of diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma syndrome, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer, wherein the composition is used to modulate dendritic cell function (e.g., thereby treating defective dendritic cell function).

[0098] In one example, a subject with defective dendritic cell function is a subject with decompensated cirrhosis. For example, a subject with defective dendritic cell function could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0099] As used herein, “regulating dendritic cells” refers to controlling dendritic cells. This encompasses, for example, controlling the number of dendritic cells, controlling dendritic cell populations, and controlling dendritic cell function. Dendritic cell function can be controlled by increasing or decreasing dendritic cell activity, for example, by stimulating a desired subset of dendritic cells (to increase the desired dendritic cell activity of these cells), and / or by increasing the (relative) abundance of desired cells (to increase the number of cells with the desired dendritic cell activity). In the context of this invention, it is desirable to regulate dendritic cell function by increasing the level of plasmacytoid dendritic cells.

[0100] As used herein, the term "level" refers to the quantity of a specified cell type (e.g., plasma cell-like dendritic cells). This level can be the absolute quantity of cells and / or the relative quantity of the specified cells. The relative quantity of the specified cells may be relative to other cells in the subject's body or from a sample from the subject. These "other cells" can be, for example, all white blood cells, or specific cell subpopulations, such as dendritic cells. The relative quantity can be, for example, a percentage, fraction, or ratio. In the context of relative quantities, it should be understood that the relative quantity of the specified cells can be changed (e.g., increased or decreased) while the absolute quantity of the specified cells may or may not be changed. Similarly, the absolute quantity of the specified cells can be changed (increased or decreased) while the relative quantity may or may not be changed.

[0101] Methods for determining whether the level of plasmacytoid dendritic cells is increased are well known. Examples of such methods are provided in the Examples section below.

[0102] Plasma cell-like dendritic cells (pDCs) play a unique role in connecting innate and adaptive immunity. They possess a lymphoid shape and plasma cell morphology, with extensive endoplasmic reticulum, multiple mitochondria, and small Golgi bodies. pDCs have poor antigen-presenting capacity, particularly for exogenous antigens, but can acquire antigen-presenting cell function upon activation and express co-stimulatory molecules that can guide T cells toward specific functional subsets. Once activated by a TLR, the expression of co-stimulatory molecules is induced in all pDCs, thus conferring these cells with T cell initiation properties. Although pDCs can be activated via different cell surface receptors and cytosol nucleic acid sensors, nucleic acid sensing via TLR7 and TLR9 appears to be the primary activation mode for IFN production in these cells. Signaling via these two TLRs leads to the rapid and massive production of all type I and type III IFNs, triggering the induction of IFN-stimulated genes (ISGs), many of which possess antiviral properties. Due to these properties, it is hypothesized that a key function of pDCs is as antiviral cells. Activation of pDCs and the production of IFN-1 are important for antiviral responses and can promote tissue repair. However, chronic or long-term persistent activation of these cells (as seen in autoimmunity and persistent viral infections) may lead to or contribute to impaired immunity and disease progression.

[0103] Therefore, as used herein, "deficient pDC" or "deficient pDC function" can refer to pDCs that are chronically or persistently activated. Deficient pDC function also refers to pDCs that reduce the production or expression of IFN molecules. In some examples, a deficient pDC can refer to a pDC that overproduces IFN molecules, such as IFN-I in cases of systemic lupus erythematosus and psoriasis. This can lead to abnormal T cell production. In some examples, deficient pDCs may produce immunosuppressive mediators such as OX40L and ICOSL, which may at least partially suppress other immune cells.

[0104] Markers of dysfunctional pDCs can be detected by determining the levels of cytokines such as IFN molecules. Other markers of pDC dysfunction include elevated Tim-3 expression.

[0105] Also provided are corresponding methods for modulating dendritic cell function (and / or preventing viral infection) in subjects, wherein a composition comprising human albumin is administered to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells. Also provided are corresponding methods for treating defective dendritic cell function in subjects in need.

[0106] Regulation of monocytes

[0107] The inventors have demonstrated in this paper that albumin treatment specifically increases the abundance of intermediate HAVCR2+ monocytes in subjects. Figure 2D-2F The data presented in this article indicate that albumin can regulate monocytes (particularly by increasing the level of intermediate HAVCR2+ monocytes) and alter transcription in monocytes, thereby changing monocyte function.

[0108] Therefore, this article provides a composition comprising human albumin for modulating monocytes in a subject, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Such compositions can be used to modulate monocyte function in a subject. Intermediate HAVCR2+ monocytes are monocytes that suppress inflammatory responses from other immune cells, such as classical monocytes and CD4 T cells (Front. Immunol 2016; 7, 229). HAVCR2 (also known as Tim 3) is an immune checkpoint molecule. Therefore, the compositions provided herein can be used to suppress inflammatory responses in a subject. Therefore, this article also provides a composition comprising human albumin for suppressing inflammatory responses in a subject, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

[0109] The compositions provided herein are particularly useful when administered to subjects suffering from systemic inflammatory response syndrome (SIRS). Subjects with SIRS may have defective monocyte function (particularly insufficient levels of intermediate HAVCR2+ monocytes). Advantageously, the compositions provided herein can be used to increase the level of intermediate HAVCR2+ monocytes in these subjects and are therefore particularly useful for suppressing the inflammatory response in subjects suffering from SIRS. Therefore, this article provides a composition comprising human albumin for modulating monocytes (e.g., modulating monocyte function) in subjects suffering from SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Thus, the compositions provided herein can be used to treat SIRS in subjects by modulating monocytes (e.g., modulating monocyte function). This article also provides a composition comprising human albumin for suppressing the inflammatory response in subjects suffering from SIRS, wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Thus, the compositions provided herein can be used to treat SIRS in subjects by suppressing the inflammatory response.

[0110] Elsewhere in this article, examples of subjects with SIRS are described. These examples are equally applicable to this aspect.

[0111] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or a subject with ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0112] This article also provides a composition comprising human albumin for modulating monocytes (e.g., modulating monocyte function) in a subject with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Therefore, the composition provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in a subject by modulating monocytes (e.g., modulating monocyte function). Furthermore, this article also provides a composition comprising human albumin for suppressing an inflammatory response in a subject with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Therefore, the compositions provided herein can be used to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by inhibiting the inflammatory response.

[0113] The compositions provided herein are also useful when administered to subjects with defective monocyte function, such as defective intermediate HAVCR2+ monocyte function. Advantageously, the compositions provided herein can be used to increase the level of intermediate HAVCR2+ monocytes in these subjects, and therefore can be specifically used to suppress the inflammatory response in subjects with defective monocyte function. Therefore, this document provides a composition comprising human albumin for treating defective monocyte function in subjects in need. In this example, human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. This document also provides a composition comprising human albumin for suppressing the inflammatory response in subjects with defective monocyte function. In this example, human albumin is administered / given to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

[0114] Those skilled in the art can readily identify examples of subjects with defective monocyte function. For instance, defective monocyte function may be associated with diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, defective monocyte function may be present in subjects with diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, the compositions comprising human albumin described herein can be used to treat defective monocyte function in a selection of diseases including: SIRS, decompensated cirrhosis, acute liver failure, pre-acute chronic liver failure, chronic-on-acute liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer (in other words, the compositions can be used to treat defective monocyte function in the aforementioned diseases).

[0115] Therefore, this article also provides a composition comprising human albumin for the treatment of diseases selected from: SIRS, decompensated cirrhosis, acute liver failure, pre-acute liver failure, chronic-on-acute liver failure, severe sepsis, septic shock, severe multiple trauma syndrome, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer, wherein the composition is used to modulate monocyte function (e.g., thereby treating defective monocyte function).

[0116] In one example, a subject with defective monocyte function is a subject with decompensated cirrhosis. For example, a subject with defective monocyte function could be a subject with pre-ACLF decompensated cirrhosis (e.g., early pre-ACLF decompensated cirrhosis) or ACLF decompensated cirrhosis. In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0117] As used herein, “monocyte regulation” refers to the control of monocytes. This encompasses, for example, controlling the number of monocytes, controlling the monocyte population, and controlling monocyte function. Monocyte function can be controlled by increasing or decreasing monocyte activity, for example, by stimulating a desired subset of monocytes (to increase the desired monocyte activity of these cells), and / or by increasing the (relative) abundance of desired cells (to increase the number of cells with the desired monocyte activity). In the context of this invention, it is desirable to regulate monocyte function by increasing the level of intermediate HAVCR2+ monocytes.

[0118] As used herein, the term "level" refers to the quantity of a specified cell type (e.g., intermediate HAVCR2+ monocytes). This level can be the absolute quantity of cells and / or the relative quantity of the specified cells. The relative quantity of the specified cells can be relative to other cells in the subject's body or from a sample from the subject. These "other cells" can be, for example, all white blood cells, or a specific cell subset, such as monocytes. The relative quantity can be, for example, a percentage, a fraction, or a ratio. In the context of relative quantities, it should be understood that the relative quantity of the specified cells can be changed (e.g., increased or decreased) while the absolute quantity of the specified cells may or may not be changed. Similarly, the absolute quantity of the specified cells can be changed (increased or decreased) while the relative quantity may or may not be changed.

[0119] Methods for determining whether the level of intermediate HAVCR2+ monocytes is increased are well known. Examples of such methods are provided in the Examples section below.

[0120] Also provided are corresponding methods for modulating monocyte function (and / or suppressing inflammatory responses) in subjects, wherein a composition comprising human albumin is administered to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Also provided are corresponding methods for treating defective monocyte function in subjects in need.

[0121] Composition

[0122] The compositions provided herein contain human albumin.

[0123] Those skilled in the art will understand that the term "albumin" as used herein encompasses proteins having the same and / or very similar tertiary structure as human serum albumin (HSA) or its HSA domains and properties similar to those of HSA or related domains. The term albumin includes variants and / or derivatives, such as fusions and / or conjugates of albumin or albumin variants. The term "variant" refers to a polypeptide derived from parental albumin that contains alterations at one or more positions, namely substitutions, insertions, and / or deletions. Substitution refers to replacing an amino acid occupying a position with a different amino acid; deletion refers to removing an amino acid occupying a position; and insertion refers to adding 1-3 amino acids adjacent to an amino acid occupying a position. The altered polypeptide (variant) can be obtained through artificial intervention by modifying the polynucleotide sequence encoding naturally occurring (wild-type) albumin.

[0124] In a preferred embodiment of the invention, the albumin is human albumin, preferably human albumin purified from human plasma (also referred to herein as human plasma-derived albumin, human serum albumin, or serum albumin). Examples of commercially available human serum albumin include Albutein® and Plasbumin® (Grifols). In some examples, the albumin is recombinant human albumin. Numerous examples of recombinant human albumin are known in the art, including: commercially available recombinant human albumin such as Recombumin® (Albumedix), Cellastim S® (InVitria), and Albagen® (AlbuminBioscience), and human albumin expressed in plants (e.g., rice) and yeasts (e.g., Saccharomyces cerevisiae and Pichia pastoris).

[0125] As used herein, the term “recombinant” refers to a biomolecule, such as a gene or protein, that (1) has been removed from its natural environment, (2) is not associated with all or part of the polynucleotides of genes found in nature, (3) is operatively linked to polynucleotides not found in nature, or (4) is not found in nature. The term “recombinant” can also refer to cloned DNA isolates, chemically synthesized polynucleotide analogs, or polynucleotide analogs biosynthesized via heterologous systems, as well as proteins and / or mRNA encoded by such nucleic acids. In some embodiments, albumin is recombinant albumin, such as recombinant human albumin.

[0126] As used herein, the term "human plasma-derived" refers to biomolecules, such as genes or proteins, obtained from a standard blend of human plasma from donors. In some embodiments, the term "human plasma-derived" is used to refer to human plasma-derived albumin.

[0127] The compositions described herein are intended for administration to a subject. Therefore, they may be referred to as pharmaceutical compositions. These compositions may include additional active agents (in addition to the human albumin described herein, such as serum albumin or recombinant human albumin), provided that the presence level (amount, concentration, or dose) of the human albumin is sufficient to achieve the stated function. In other words, the compositions described herein contain a level (amount, concentration, or dose) of human albumin sufficient to modulate immune cells in a subject in the manner described.

[0128] The pharmaceutical composition may contain the human albumin described herein, as well as pharmaceutically acceptable excipients, adjuvants, diluents, and / or carriers.

[0129] The composition may conventionally contain pharmaceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, supplemental immune enhancers (such as adjuvants and cytokines), and optionally other therapeutic agents or compounds.

[0130] As used herein, “pharmaceutical acceptable” means a material that is not biologically or otherwise undesirable, i.e., that the material can be administered to an individual in conjunction with human albumin without causing any adverse biological effects or interacting in a harmful manner with any other component of a pharmaceutical composition containing the material.

[0131] Excipients are natural or synthetic substances formulated with an active ingredient (such as human albumin as described herein) to increase the volume of the formulation or to impart therapeutic enhancement to the active ingredient in the final dosage form, such as promoting drug absorption or dissolution. Excipients are also useful in the manufacturing process to aid in handling the associated active substance, such as by promoting powder flowability or non-stick properties, and also to contribute to in vitro stability, such as preventing denaturation within the expected shelf life. Pharmaceutically acceptable excipients are well known in the art. Therefore, suitable excipients can be readily identified by those skilled in the art. For example, suitable pharmaceutically acceptable excipients include water, saline, aqueous glucose solutions, glycerol, ethanol, etc.

[0132] An adjuvant is a pharmacological and / or immunological agent that alters the action of other agents in a formulation. Pharmaceutically acceptable adjuvants are well known in the art. Therefore, suitable adjuvants can be readily identified by those skilled in the art.

[0133] A diluent is a diluting agent. Pharmaceutically acceptable diluents are well known in the art. Therefore, those skilled in the art can readily identify suitable diluents.

[0134] The carrier is non-toxic to the recipient at the used dosage and concentration and is compatible with other ingredients in the formulation. The term "carrier" refers to a natural or synthetic organic or inorganic ingredient with which the active ingredient is combined to facilitate application. Pharmaceutically acceptable carriers are well known in the art. Therefore, suitable carriers can be readily identified by those skilled in the art.

[0135] As used herein, the term "subject" refers to an individual who has or is at risk of having a particular condition, symptom, or illness, such as a human. A subject can be a patient, i.e., a subject who requires treatment according to the present invention. The subject may have already received treatment for that condition, symptom, or illness. Alternatively, the subject may not have received treatment prior to treatment according to the present invention.

[0136] The compositions described herein can be administered to a subject via any conventional route, including injection or gradual infusion over time. Administration can be made, for example, via a route selected from: intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

[0137] The compositions described herein can be in any form suitable for the above-described administration method. Those skilled in the art can determine the appropriate dosage of the compositions described herein.

[0138] The compositions described herein are intended for administration in an effective amount. An "effective amount" is the amount that, alone or in combination with other doses, produces the desired (therapeutic or non-therapeutic) response. The effective amount used will depend, for example, on the therapeutic (or non-therapeutic) purpose, route of administration, and the patient / subject's condition. For instance, the appropriate dose of the compositions of the present invention for a given patient / subject will be determined by the attending physician (or the person administering the composition), taking into account various factors known to alter the effects of the compositions of the present invention, such as the severity and type of hematologic malignancies, weight, sex, diet, time and route of administration, other medications, and other relevant clinical factors. Dosage and schedule can vary depending on the patient / subject's specific condition, illness or symptoms, and overall health.

[0139] The pharmaceutical compositions described herein are advantageously presented in unit dosage forms.

[0140] In one example, human albumin (e.g., serum albumin or recombinant human albumin) is used as part of a multiple-dose regimen administered at multiple intervals. Those skilled in the art can determine suitable multiple-dose regimens. For example, a multiple-dose regimen may comprise multiple partial doses administered at equal intervals of about 1 day to about 30 days. In one example, a multiple-dose regimen may comprise multiple partial doses administered at unequal intervals of about 1 day to about 30 days. In one example, a multiple-dose regimen may comprise two or more doses until the total cumulative dose is reached. In some examples, human albumin may be administered at equal or unequal intervals of about 1 day to 6 days.

[0141] The compositions described herein contain appropriate levels (amount, concentration, or dose) of human albumin. In the context of any dosing regimen described herein, when used as part of a multiple dosing regimen (e.g., as part of any multiple dosing regimen specifically described herein), the human albumin in the composition may be administered at a single dose of about 5 g / interval to about 500 g / interval. For example, when used as part of a multiple dosing regimen (e.g., as part of any multiple dosing regimen specifically described herein), the human albumin in the composition may be administered at a single dose of about 20 g / interval to about 200 g / interval. For example, 40 g / interval. For example, 40 g / day.

[0142] In specific examples, a multiple-dose regimen may comprise administering multiple fractional doses of the composition described herein at equal intervals (equal intervals) from about 1 day to about 30 days, wherein the human albumin in the composition is used to administer a single dose of about 5 g / interval to about 500 g / interval (e.g., about 20 g / interval to about 200 g / interval). In another specific example, a multiple-dose regimen may comprise administering multiple fractional doses of the composition described herein at unequal intervals (unequal intervals) from about 1 day to about 30 days, wherein the human albumin in the composition is used to administer a single dose of about 5 g / interval to about 500 g / interval (e.g., about 20 g / interval to about 200 g / interval). In yet another specific example, a multiple-dose regimen may comprise administering the composition described herein two or more times until a total cumulative dose is reached, wherein the human albumin in the composition is used to administer a single dose of about 5 g / interval to about 500 g / interval (e.g., about 20 g / interval to about 200 g / interval). In some examples, a multiple-dose regimen may comprise administering about 40 g / interval of human albumin. In some examples, a multiple-dose regimen may include administering approximately 40 g / day of human albumin. In some examples, a regimen may include administering approximately 40 g / interval of human albumin over a period of 2 to 6 days. For example, a regimen may include administering approximately 40 g / day of human albumin over a period of 2 to 6 days. In some examples, a regimen may include administering approximately 40 g / day of human albumin over a median period of 2.5 days.

[0143] In each of these examples, the dosing interval can be every 15 days or less. For example, the dosing interval can be every 10 days or less.

[0144] In a specific example, the composition described herein contains human albumin at a concentration of about 4% to 25% (w / v) (e.g., about 20% (w / v)). In other words, the dosages and dosing regimens provided above can be achieved using a composition containing human albumin at a concentration of about 4% to 25% (w / v) (e.g., about 20% (w / v)).

[0145] The compositions described herein are particularly useful for treating subjects. The terms "treatment" or "treating" refer to any treatment of a disease or condition in a subject (such as a mammal), including: prevention or avoidance of the disease or condition, i.e., causing the absence of clinical symptoms; inhibition of the disease or condition, i.e., stopping or suppressing the development of clinical symptoms; and / or relief of the disease or condition, i.e., causing the remission of clinical symptoms. In some examples, the term "treatment" is used to treat defective immune cell function. In some embodiments, the term "treatment" is used to treat diseases selected from: systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure, acute-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

[0146] Those skilled in the art will understand that in human medicine, it is not always possible to distinguish between “prevention” and “inhibition” because the eventual inducing event may be unknown, latent, or the patient may not be diagnosed until long after one or more events have occurred. Therefore, as used herein, the term “prevention” is intended to encompass “prevention” and “inhibition” as defined herein, as an element of “treatment.”

[0147] As used in this article, systemic inflammatory response syndrome (SIRS) refers to an inflammatory state resulting from infection or non-infectious causes. SIRS is a serious condition associated with systemic inflammation, organ dysfunction, and organ failure. It is a subset of a cytokine storm, in which various cytokines are abnormally regulated. SIRS is also closely associated with sepsis, in which patients meet the criteria for SIRS and have a suspected or confirmed infection. The following criteria can be used to identify subjects with SIRS: In adults, SIRS manifestations include, but are not limited to: (i) Body temperature below 36 °C (96.8 °F) or above 38 °C (100.4 °F) (ii) Heart rate greater than 90 beats per minute (iii) Rapid breathing (high respiratory rate), greater than 20 breaths per minute; or arterial carbon dioxide partial pressure less than 4.3 kPa (32 mmHg). (iv) White blood cell count less than 4000 cells / mm³ (4 x 10⁻⁶) 9 (12,000 cells / L) or more than 12,000 cells / mm 3 (12 x 10) 9(cells / L); or the presence of more than 10% immature neutrophils (band-like pattern). A band-like pattern greater than 3% is called bandemia or "left shift".

[0148] When two or more of these criteria are met, a patient may be diagnosed with SIRS, regardless of whether there is evidence of infection. Patients with SIRS and acute organ dysfunction may be referred to as having "severe SIRS".

[0149] In children, the SIRS criteria have been modified as follows: (i) A heart rate that is more than two standard deviations above the normal range for its age, or an unexplained sustained increase in heart rate for more than 30 minutes to 4 hours, without any stimuli such as pain or drug administration. In infants, this also includes a heart rate that is below the 10th percentile for its age, or an unexplained sustained decrease for more than 30 minutes, without vagal stimulation, beta-blockers or congenital heart disease.

[0150] (ii) Body temperature measured orally, rectally, via a Foley catheter probe or via a central venous catheter probe is below 36°C or above 38.5°C.

[0151] (iii) A respiratory rate that is more than two standard deviations above the normal value for age, or the need for mechanical ventilation that is not related to neuromuscular disease or drug-induced anesthesia.

[0152] (iv) White blood cell count is elevated or decreased relative to age, unrelated to chemotherapy, or band-like pattern plus other immature patterns exceeding 10%.

[0153] In pediatric patients, an abnormal body temperature or white blood cell count is required to be considered a case of SIRS.

[0154] SIRS may be associated with subjects suffering from a condition selected from the following: decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure (ACLF), ACLF, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, SIRS may be present in subjects suffering from a condition selected from the following: decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure (ACLF), ACLF, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic lymphohistiocytosis (HLH), heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer. Therefore, the compositions comprising human albumin described herein can be used to treat SIRS in a selection of diseases including: decompensated cirrhosis, acute liver failure, pre-acute-on-chronic liver failure (ACLF), acute-on-chronic liver failure (ACLF), severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer (in other words, the compositions can be used to treat the SIRS aspect of these diseases). Methods for identifying whether a subject has one or more of these diseases are well known.

[0155] In one example, the subject is a patient with sepsis or a liver condition. Therefore, a subject with SIRS may have SIRS associated with sepsis or a liver condition. Examples of patients with sepsis include those with severe sepsis or septic shock. Examples of patients with liver conditions include those with decompensated cirrhosis, acute liver failure, pre-acute chronic-to-acute liver failure (ACLF), and / or acute chronic-to-acute liver failure (ACLF).

[0156] In one example, the subject had decompensated cirrhosis. Decompensated cirrhosis is defined as an acute deterioration of liver function in a patient with cirrhosis, characterized by recent onset of ascites, hepatic encephalopathy, gastrointestinal bleeding, or any combination thereof. Decompensated cirrhosis is also referred to herein as “acute decompensated cirrhosis,” “acute decompensated cirrhosis,” or “decompensated liver disease,” and these terms are used interchangeably.

[0157] In subjects with acute decompensated cirrhosis, those with chronic-on-acute liver failure (ACLF), a syndrome characterized by severe systemic inflammation, single multi-organ dysfunction / failure, and an extremely high prevalence of bacterial infections, were more significantly affected than those without ACLF (Moreau et al., 2013, Gastroenterology, 144:1426-37, 1437.e1-9; Arroyo et al., 2020, N Engl J Med, 382:2137-2145) (Weiss et al., 2021, Frontiers in Immunol, vol. 12, art. 699563). Furthermore, the PREDICT study recently identified a subset of subjects with severe systemic inflammation but without ACLF who were at high risk of developing ACLF and dying during hospitalization or within weeks of discharge (Trebicka et al., 2020, J Hepatol, 73:842-854). The participants in this subset were classified as having acute decompensated cirrhosis in the pre-ACLF stage.

[0158] Pre-ACLF is defined as a group of patients with acute decompensated cirrhosis who did not have ACLF at admission but developed ACLF during hospitalization or within 3 months of admission. Pre-ACLF can be divided into two distinct phenotypes: "early" pre-ACLF, defined as patients who develop ACLF during their initial hospitalization; and "delayed" pre-ACLF, defined as patients who develop ACLF between discharge from their initial hospitalization and the end of the 3-month follow-up period. Patients hospitalized for "early" pre-ACLF exhibit strong systemic inflammation, while this characteristic is more mild (mild) in patients with "slowly progressive" pre-ACLF.

[0159] Subjects with acute decompensated cirrhosis and ACLF are also referred to as ACLF subjects or ACLF patients in this article. Subjects with pre-ACLF acute decompensated cirrhosis are also referred to as pre-ACLF subjects or pre-ACLF patients in this article.

[0160] In one example, the subject is an ACLF subject or a pre-ACLF subject. In other words, the subject may have ACLF decompensated cirrhosis (also referred to as acute ACLF decompensated cirrhosis in this article) or pre-ACLF decompensated cirrhosis (also referred to as pre-acute ACLF decompensated cirrhosis in this article). In one example, pre-ACLF decompensated cirrhosis is early-stage pre-ACLF decompensated cirrhosis.

[0161] In one example, the subject suffered from acute liver failure. Acute liver failure is characterized by acute liver injury, hepatic encephalopathy (altered mental status), and elevated prothrombin time / international normalized ratio (INR). It is also known as fulminant hepatic failure, acute hepatic necrosis, fulminant hepatic necrosis, and fulminant hepatitis.

[0162] Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. For example, Singleton and Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d Ed., John Wiley and Sons, NY (1994); and Hale and Marham, The Harper Collins Dictionary of Biology, Harper Perennial, NY (1991) provide general dictionaries for those skilled in the art of the many terms used in this invention. Although any methods and materials similar to or equivalent to those described herein may be used to practice this invention, preferred methods and materials are described herein. Therefore, the terms defined below will be described more fully by reference to the entire specification. Furthermore, as used herein, the singular terms “a,” “an,” and “the” include plural references unless the context clearly indicates otherwise. Unless otherwise stated, nucleic acids are written from left to right in a 5' to 3' orientation; amino acid sequences are written from left to right in an amino to carboxyl orientation. It should be understood that the present invention is not limited to the specific methods, protocols and reagents described, as these may vary depending on the context in which they are used by those skilled in the art.

[0163] Various aspects of the invention are illustrated by the following non-limiting examples.

[0164] Example

[0165] Example 1 - Albumin therapy for gene expression in blood immune cells of patients with acute decompensated cirrhosis The effects of characteristics (the effects of albumin on blood immune cells)

[0166] background

[0167] Assessing whole-genome RNA expression in blood is a valuable tool for simultaneously studying the characteristics of different types of circulating immune cells. 5,19-21The inventors now report results from the Discovery study, an analysis of whole blood RNA-seq data obtained twice from 49 patients in the PREDICT study; the first time when these patients were in the pre-ACLF stage (at admission), and the second time when they developed the ACLF stage a few days after admission during their initial hospitalization. Therefore, all patients had an “early” form of pre-ACLF. Because 30 patients received albumin therapy during progression to ACLF, while the remaining 19 did not, the Discovery study was the first attempt to capture the in vivo effects of albumin on gene signatures associated with blood immune cells. The longitudinal design and demonstrated similarity of the systemic inflammatory intensity of the “early” pre-ACLF stage to ACLF should minimize inter-individual variability in gene expression, which would otherwise be difficult to capture in vivo effects of albumin on blood immune cells.

[0168] Surprisingly, the Discovery study found that patients treated with albumin showed specific increases in gene signatures primarily associated with B cells, plasma cells, and neutrophils. These findings prompted the inventors to conduct additional studies, including with new patients with acute decompensated cirrhosis, to further investigate the effects of albumin on cells that make up freshly isolated peripheral blood mononuclear cells (PBMCs) and on freshly isolated blood neutrophils in vitro.

[0169] method

[0170] Patients and study design

[0171] All patients (i.e., 64 patients) had acute decompensated cirrhosis but not ACLF at enrollment. Written informed consent was provided by each patient to participate. Each study described below had previously been approved by the Ethics Committee of the Barcelona Hospital Clinic, Spain.

[0172] Discovery Research

[0173] The inventors first considered PREDICT research. 3The 49 patients were carefully selected based on the following criteria: i) All patients had “early” pre-ACLF at enrollment (time 1 [T1]) because they developed ACLF during their initial hospitalization (time 2 [T2]; data not shown). For simplicity, the inventors have hereafter used the term “pre-ACLF” instead of “early pre-ACLF”. The inventors hypothesized that using longitudinal data to study patients exhibiting the most severe form of acute decompensated cirrhosis would limit the impact of inter-individual variability on the assessment of albumin effects. ii) Whole blood RNA-seq data were obtained at T1 and at T2, as pre-defined in the protocol, in all patients. iii) None of the 49 patients had received albumin prior to T1. iv) 30 of the 49 patients received albumin between T1 and T2 and formed the albumin group, while the remaining 19 patients did not receive albumin between T1 and T2 and formed the no-albumin group (data not shown). Notably, the inventors also obtained clinical blood cell counts and whole blood RNA-seq results (data not shown) from 10 age-matched healthy subjects (HS). The inventors analyzed longitudinal whole blood RNA-seq data from 49 patients for two purposes. The first was to assess whether the transcriptomic characteristics of the 49 patients studied in the pre-ACLF stage differed from those in the same patients when ACLF progressed, regardless of albumin administration. Indeed, unlike the previously described whole blood transcriptomic characteristics of ACLF, 5 The whole blood transcriptome profile of pre-ACLF has not yet been investigated. The second primary objective is to assess whether albumin affects the RNA profile of circulating immune cells by analyzing the blood transcriptome in both albumin- and albumin-free groups.

[0174] Additional research

[0175] The findings of the Discovery study prompted the inventors to recruit nine new patients with acute decompensated cirrhosis whose peripheral blood mononuclear cells (PBMCs) were freshly isolated to investigate the in vitro effects of albumin on the cells that make up PBMCs using single-cell RNA-seq (scRNA-seq). Additionally, neutrophils were freshly isolated from six new patients in an additional group to investigate the in vitro effects of albumin on the antimicrobial function of major neutrophils.

[0176] RNA preparation for whole blood RNA-seq

[0177] As previously described, 5,19RNA was isolated from blood stored in Tempus tubes using the Tempus™ Spin RNA Isolation Kit (Reference 43802, Applied Biosystems, Foster City, CA). Importantly, using the HUGO Genome Nomenclature Committee (an approved resource for naming human genes), the inventors devised a comprehensive protocol that included not only protein-coding genes but also genes from other locus types, such as immunoglobulin genes and T-cell receptor genes, resulting in a total of 14,615 genes for analysis. Genes derived from sex chromosomes and mitochondrial DNA were not included in the analysis.

[0178] Whole blood RNA-Seq analysis

[0179] Assess differential gene expression.

[0180] Differentially expressed genes (DEGs) are defined as having two criteria: an absolute fold change (FC) greater than 1.5 and a regulatory P-value less than 0.05.

[0181] Immune cell characteristics were assessed using SingleR R software.

[0182] Using SingleR software 22 Whole blood RNA-seq data from study participants were compared with a reference dataset (GSE107011) containing 114 human bulk RNA-seq samples from sorted immune cell populations from four healthy subjects. 23 The reference dataset contains 114 human RNA-seq samples, annotated for 10 major immune cell types. This dataset (GSE107011) also contains additional samples annotated for 29 finer immune cell types. The SingleR workflow first compares the whole blood RNA-seq data with the 10 reference major immune cell types, and then with the 29 finer immune cell types.

[0183] Analysis of gene set enrichment

[0184] Use gene expression quantification analysis (QuSAGE) methods, such as those implemented in the QuSAGE package. 24 Gene set enrichment analysis was performed using 346 blood transcriptome modules (BTMs) as a gene set. BTMs are a set of gene modules developed through large-scale network integration of publicly available human blood transcriptomes. 20QuSAGE provides an activity score for each gene set in each comparison (e.g., ACLF pre-comparison with healthy subjects; T2 vs. T1 in the albumin group). A BTM is identified as differentially expressed when its QuSAGE activity score has a false discovery rate (FDR) < 0.05 or P < 0.05 (where appropriate).

[0185] Analysis of shared genes and shared BTM

[0186] When the DEG or the differentially expressed BTM is sign-consistent, the DEG or the differentially expressed BTM is defined as shared between the two comparisons.

[0187] ScRNA-seq in PBMC

[0188] Peripheral venous blood (8 ml) was collected from 9 patients and 4 age-matched healthy subjects via venipuncture and placed into EDTA-coated sterile, pyrogen-free tubes (Becton Dickinson, East Rutherford, NJ). PBMCs were immediately isolated as previously described. 25 PBMC at 1.5x10 6 The cells were seeded at a density of 100 cells / ml and incubated for 2 hours at 37°C in a 5% CO2 incubator with albumin (Albutein®, Grifols, Barcelona, ​​Spain) (15 mg / ml) or medium. At the end of the culture, the cells were rapidly (within 30 min) transferred to ice and sent to CNAG’s single-cell genomics platform.

[0189] Experiments in neutrophils

[0190] Neutrophils were isolated from peripheral venous blood (20 ml) obtained from 6 patients and 5 age-matched healthy subjects using the Ficoll-Hypaque method. The isolated neutrophils were immediately used in various experiments to assess the major antimicrobial functions of neutrophils, such as degranulation assays and phagocytosis assays.

[0191] result

[0192] Characterization of pre-ACLF at T1 and ACLF at T2 in 49 patients in the Discovery Cohort

[0193] Clinical features

[0194] The inventors first characterized all patients at T1 (pre-ACLF) and T2 (progression to ACLF). Characterization was similar in terms of standard laboratory values ​​(including clinically differential blood cell counts of neutrophils, monocytes, and lymphocytes; data not shown) and blood levels of inflammatory markers (data not shown). Notably, blood levels of inflammatory markers were elevated in both pre-ACLF and ACLF compared to corresponding levels in healthy subjects (data not shown). The only two differences were the presence of organ failure in ACLF at T2 but not in pre-ACLF at T1, which was consistent with expectations, and higher white blood cell counts in ACLF compared to pre-ACLF (data not shown). Compared to healthy subjects, pre-ACLF and (as expected) ACLF... 5 The characteristics of these findings were leukocytosis, neutropenia, and lymphopenia (data not shown). Notably, the inventors discovered a strong positive correlation between clinical blood cell counts and blood cell counts inferred from RNA-seq (data not shown), suggesting that RNA-seq analysis of whole blood RNA can provide information relevant to criterion-standard clinical blood cell count measurements.

[0195] Next, the inventors considered the characteristics at T1 and T2 in patients in the albumin group and the albumin-free group. In the albumin group, the median time between T1 and T2 for blood collections used for RNA-seq was 11.5 days (interquartile range [IQR], 6.2 to 18.7). The median duration of albumin treatment was 2.5 days (IQR, 2 to 6), and the albumin dose was 40.0 g daily (IQR, 22.5 to 57.5). The time between the last albumin administration and the subsequent blood collection for RNA-seq was 1.5 days (IQR, 0.0 to 2.7), indicating that the inventors explored patients shortly after albumin administration. Albumin was administered according to standard medical indications. In the albumin-free group, the median time between T1 and T2 was 29 days (IQR, 16 to 52).

[0196] In both the albumin and non-albumin groups, progression from pre-ACLF (T1) to ACLF (T2) was associated with a significant increase in MELD scores. However, apart from the increased serum creatinine at T2, there were no significant changes within the groups in the remaining longitudinally collected standard laboratory data and circulating levels of 14 inflammatory mediators (Table 1). However, some intergroup differences were observed. The delay between T1 and T2 was shorter in the albumin group than in the non-albumin group. At both T1 and T2, serum levels of C-reactive protein and interleukin-6 were higher in the albumin group. A higher percentage of patients in the albumin group died within 28 and 90 days compared to those in the non-albumin group; this difference was expected given the higher prevalence of SBP and HRS-AKI in the albumin group.

[0197] Transcriptional characteristics of all patients at the pre-ACLF stage (T1) and ACLF stage (T2)

[0198] Analyzing the DEGs in the two comparisons (pre-ACLF vs. healthy subjects and ACLF vs. healthy subjects), as expected, the inventors observed a strong similarity between the two disease stages. Consequently, the number of DEGs was high in both comparisons: 4615 DEGs in pre-ACLF compared to healthy subjects, and 4529 DEGs in ACLF compared to healthy subjects. Of the 4615 DEGs associated with pre-ACLF, 3929 (85%) overlapped with the DEGs assigned to ACLF (data not shown). Furthermore, when the inventors compared the effect size change (log2 fold change [FC]) in pre-ACLF vs. healthy subjects with the effect size change in ACLF vs. healthy subjects, they observed a high degree of consistency in the magnitude of change between the two signatures (data not shown).

[0199] Next, the inventors used QuSAGE to analyze the differential expression of blood gene modules (BTMs). BTMs are sets of genes associated with blood cells (e.g., plasma cells, immunoglobulins (M156.1)) or biological functions (e.g., muscle contraction, SRF targets (M195)). Of the 258 annotated BTMs, 20The total number of differentially expressed (DE) BTMs (FDR < 0.05) was 190 in pre-ACLF versus healthy subjects and 186 in ACLF versus healthy subjects (data not shown), indicating extensive changes in the blood transcription module space in both pre-ACLF and ACLF. Notably, only 20 BTMs showed specific differential expression relative to healthy subjects in either pre-ACLF (12 modules) or ACLF (8 modules), while 179 DE BTMs were shared, i.e., DE modules with consistent signs in both comparisons (Table SX). Of these shared DE gene modules, 108 were upregulated and 71 were downregulated. The top 20 gene modules of shared upregulated BTMs and the top 20 gene modules of shared downregulated BTMs were documented (data not shown). The shared upregulated BTMs are associated with innate immunity, including BTMs related to TLR and inflammatory signaling, interferon-α responses, and innate immune cells such as neutrophils, monocytes, and dendritic cells. Shared downregulated BTM was associated with T cells, NK cells, and antigen presentation. These results are consistent with those obtained when analyzing the genetic signatures of 10 major immune cell types and 29 fine immune cell types using SingleRR software. Overall, the analysis of whole blood RNA-seq data using QuSAGE and SingleRR software provided consistent results highlighting the similarity of the blood transcriptional module landscape in pre-ACLF and ACLF; both disease stages are characterized by a simultaneous increase in genetic signatures associated with innate immune cells and a decrease in genetic signatures associated with adaptive immune cells.

[0200] Whole blood genetic characteristics associated with albumin therapy

[0201] The inventors studied the differential gene expression between T2 and T1 in the albumin-only and non-albumin-only groups and found that the gene characteristics of the two groups were different. In fact, 269 DEGs (92 upregulated genes and 177 downregulated genes) were specific to the albumin-only group, while 103 DEGs (64 upregulated genes and 39 downregulated genes) were specific to the non-albumin-only group; the two groups shared only 36 DEGs with consistent symbols. Figure 1A Identification of DEG in volcano maps ( ). Figure 1B This illustrates the group-specificity of upregulated and downregulated genes. For example, in the albumin group, at T2 versus T1, various immunoglobulin genes (see below) and several major neutrophil genes were upregulated. CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, MCEMP1 () Figure 1B (see above), but not in those groups that were upregulated in the albumin-free group ( Figure 1B (See the image below).

[0202] Due to the analysis of DEG ( Figure 1B This led to increased interest in immunoglobulin genes. The inventors compared the effect size changes (log2 fold changes) between T2 and T1 for each of the 125 genes (including 120 immunoglobulin genes) included in the gene set called "Immunoglobulin Complexes." They found that the effect size changes were greater in the albumin group than in the non-albumin group, with 33 immunoglobulin genes upregulated compared to only 6. Figure 1C Genes encoding the constant region of the immunoglobulin heavy chain ( IGHM , IGHG2 , IGHG3, IGHG4, IGHA2 ) were specifically upregulated in the albuminome. These findings are consistent with the results of the SingleR analysis, which showed an increase in plasmablast genetic signatures, which is albuminome-specific. Figure 1D (See the image above). Immunoglobulin genes specifically upregulated in the albuminome also include immunoglobulin light chain κ (…). IGKC ) and λ ( IGLC1 The genes specifically upregulated in the albumin-free group were those encoding the constant region of the immunoglobulin heavy chain, the V region of the variable domain of the immunoglobulin heavy chain, and the V region of the variable domains of the light chains κ and λ. The three genes specifically upregulated in the albumin-free group were those encoding the V region of the variable domain of the immunoglobulin heavy chain (Figure 10). These findings collectively indicate a widespread upregulation of immunoglobulin-encoding genes, specific to patients receiving albumin.

[0203] Next, the inventors applied QuSAGE to identify differentially expressed BTMs between T2 and T1 within each group. The inventors first observed that the number of differentially expressed BTMs was greater in the albumin group than in the albumin-free group (36 vs. 6). Only one differentially expressed BTM was specific to the albumin-free group; this BTM was upregulated and associated with the endoplasmic reticulum (ER). Figure 1E In stark contrast, 31 differentially expressed BTMs (13 downregulated and 18 upregulated) are albuminome-specific. Figure 1E The downregulated modules are associated with erythropoiesis, the cytoskeleton, or cell junctions. Figure 1E Within the albuminome-specific upregulated BTM, there are modules associated with innate immunity and inflammation, including those related to activated dendritic cells; complement and other receptors in dendritic cells (DCs); and those associated with pro-inflammatory cytokines and chemokines. Figure 1EAlbuminome-specific upregulated BTMs also include modules associated with B cells (including those related to B cell, plasma cell, and immunoglobulin enrichment) and modules associated with mismatch repair, cell cycle, and mitosis. Finally, BTMs associated with transcription factor (TF) transcriptional networks (such as c-Myc and E2F family members) are specifically upregulated in the albuminome. Figure 1E These findings suggest that the activity of these TFs is increased in the blood of patients receiving albumin. Since c-Myc and E2F family members are regulators of the cell cycle, this increased activity may explain the induction of gene modules related to the cell cycle and mitosis observed in the blood of patients receiving albumin.

[0204] It is worth noting that the characteristics of T cells and NK cells inferred by RNA-seq ( Figure 1D ) and gene modules associated with these cells, downregulated, are hallmarks of ACLF (and pre-ACLF; data not shown), and remained downregulated in patients treated with albumin. However, the inventors observed that a BTM called “mitotic cell cycle (M4.11) in stimulated CD4 T cells” was specifically upregulated in patients treated with albumin ( Figure 1E This finding indicates the presence of transcriptional activation in the CD4 T cells of these patients. Consistent with this, the inventors found no overlap between member genes of BTM associated with mitotic cell cycle in stimulated CD4 T cells (M4.11) and genes used to characterize CD4 T cells using SingleRR software. Overall, the results reveal that blood from patients who received albumin is characterized by increased features associated with immune cells (such as B cells, plasma cells, some innate immune cells, and possibly CD4 T cells), accompanied by increased features of mitosis, cell cycle activity, and increased E2F family TF activity.

[0205] ScRNA-seq of human PBMCs to investigate the in vitro effects of albumin

[0206] Results obtained from whole blood RNA-seq analysis prompted the inventors to use scRNA-seq to study the in vitro effects of albumin on human PBMCs. Before conducting the scRNA-seq experiment, fresh PBMCs from 9 patients with acute decompensated cirrhosis and 4 healthy controls were exposed to albumin (15 mg / ml) and the medium for 2 hours in vitro.

[0207] The inventors jointly analyzed 66,064 human PBMCs from patients and healthy subjects, clustering these PBMCs into three major immune cell lineages: B lymphocytes, myeloid cells, and T cells. The inventors further used typical genetic markers to sub-cluster each lineage to annotate the different cell types.

[0208] The inventors analyzed 3786 B lymphocytes (1946 from patients and 1840 from healthy subjects), identifying 10 major populations. Results from patient cells are as follows: Figure 2A As shown. Notably, the inventors described two subpopulations that yielded transitional-like B-cell profiles, characterized by: CD79B , IGHD and TCL1A (Transitional B cells) and CD55 Higher expression of (transitional B cells). The inventors observed a significant increase in the abundance of cells expressing transitional B cell-like characteristics and a decrease in naive B cells in patient cells exposed to albumin. Figure 2B and 2C However, other B-lymphocyte populations remained unchanged. The inventors further evaluated 26,898 myeloid cells (21,677 from patients and 5,053 from healthy subjects), which clustered into 13 distinct cell populations. Seven of these clusters were associated with monocytes, three with dendritic cells (DCs), and the other three included myeloid-derived suppressor cells and granulocyte-monocyte progenitor cells. Results for the obtained patient cells are as follows... Figure 2D As shown. The inventors observed that in patient cells exposed to albumin, the abundance of intermediate monocytes (HAVCR2+) and plasma cell-like dendritic cells (DCs) was significantly increased. Figure 2E and 2F The abundance of other myeloid cell groups remained unchanged.

[0209] Based on differentially expressed BTM found in the T cell compartments, the inventors also analyzed 35,380 T lymphocytes, including 21,731 CD4 T cells (12,692 from patients and 9,039 from healthy subjects), 10,072 CD8 T cells (3,483 from patients and 6,589 from healthy subjects), and 3,577 unconventional T cells (1,292 from patients and 2,285 from healthy subjects). CD4 T cells clustered into 10 distinct cell populations. Results for the obtained patient cells are as follows... Figure 2G As shown. The inventors observed that patient exposure to albumin altered the profile of CD4 T cell compartments ( Figure 2H The abundance of activated memory CD4 T cells decreased significantly, and the central memory ITGB1... +Increased CD4 T cell abundance ( Figure 2I CD8 T cells and unconventional T cells clustered into 13 distinct cell populations, but no significant changes in cell populations were observed after albumin exposure.

[0210] One of the BTMs specifically upregulated in patients who received albumin is a BTM called "mitotic cell cycle (M4.11) in stimulated CD4 T cells". Figure 1E To verify the M4.11 signature on scRNA-seq, the inventors used the Ucell package to calculate the gene signature score of this BTM in all CD4 T cells. 26 The M4.11 BTM signature showed different distribution patterns on CD4 T cells under albumin and mediator conditions, and albumin-treated cells exhibited significantly higher cell density compared to mediator-treated cells. Figure 2J ).

[0211] Furthermore, analysis of PBMCs from healthy subjects showed that albumin had similar effects on B lymphocyte and myeloid cell compartments as those in patients.

[0212] Study on the in vitro effects of albumin on neutrophils

[0213] Analysis of whole blood RNA-seq data from the albumin- and non-albumin-treated groups drew the inventors' attention to neutrophils. In fact, pairwise comparisons of whole blood RNA-seq data between T2 and T1 in the albumin- and non-albumin-treated groups revealed upregulation of nine major neutrophil genes, specific to patients receiving albumin. Figure 1B Therefore, these findings prompted the inventors to design in vitro studies in freshly isolated neutrophils. Since neutrophils provide the first line of defense against most pathogens and express fewer genes than any other white blood cell, the study focused on analyzing the effects of albumin on neutrophil function.

[0214] During the host defense response, neutrophils release myeloperoxidase (MPO), an essential antimicrobial protein that is primarily located in azurophilic or primary neutrophil granules. 27 Therefore, neutrophil degranulation can be quantified by measuring MPO activity. Figure 3 A showed that MPO activity was enhanced in the supernatant of neutrophils from patients with acute decompensated cirrhosis and healthy subjects, and incubated with serum albumin or recombinant human albumin.

[0215] Phagocytosis is another defense mechanism against pathogens, which can be assessed by determining the uptake of fluorescently labeled yeast glycan particles by neutrophils. Figure 3 B showed that neutrophils from patients with acute decompensated cirrhosis and healthy subjects, incubated with serum albumin or recombinant human albumin, exhibited enhanced phagocytosis of yeast glycans.

[0216] discuss

[0217] This study (the Discovery study) is the first to use longitudinal genomics to explore the mechanism of action of albumin in patients with acute decompensated cirrhosis. The design of this study, particularly the selection of candidates and aspects related to albumin therapy, was completed after considering known and some unpublished aspects. Patients hospitalized for ACLF are known to exhibit an extremely dynamic clinical course, improving, remaining stable, or worsening within days of admission, with significant changes in the degree of systemic inflammation. 2 Clearly, this was not a suitable candidate for this study, which aimed to evaluate the effects of albumin therapy on immune cell transcriptomics. The other three phenotypes of acute decompensated cirrhosis, including “unstable” and “stable” decompensated cirrhosis and “slowly progressive” pre-ACLF, were also not considered because they are associated with moderate systemic inflammation, which may further subside during hospitalization, and the inventors were only interested in including patients with severe systemic inflammation to better assess the potential changes in albumin-induced immune cell gene expression. For this reason, the inventors focused on patients with “early” pre-ACLF, whose level of systemic inflammation was similar to that seen in ACLF. The inventors decided to include only patients who had received albumin therapy with a clearly defined indication (percutaneous biopsy, prevention of SBP-related HRS-AKI, or treatment of HRS-AKI). 13 Although this introduces a higher degree of bias in patients receiving albumin therapy, the inventors did not select patients who received albumin within one month prior to T1, patients with excessively long intervals between two whole blood RNA-seq assessments, or patients with a delay >10 days between their last albumin dose and T2. Finally, although albumin dosage and duration of treatment vary depending on the indication, the inventors decided to analyze data from all patients regardless of albumin dosage. Of the 1273 patients enrolled in the PREDICT study, 49 met all selection criteria and were therefore investigated in this Discovery study.

[0218] The inventors first compared blood transcriptomes obtained from all patients at various time points with those obtained from healthy subjects. They observed that pre-ACLF and ACLF were characterized by increased genetic signatures associated with innate immune cells (including those associated with neutrophils, monocytes, and dendritic cells) and decreased genetic signatures associated with lymphocytes (including T cells, B cells, and NK cells). These findings suggest that myeloid-mediated innate immune activation and lymphocyte depletion occur simultaneously in the blood of patients with cirrhosis and severe systemic inflammation. The similarity between pre-ACLF and ACLF in transcriptome signatures, as well as most clinical features, standardized laboratory values, and blood cytokine levels, confirms the inventors' hypothesis that the study patients were in a relatively stable state of systemic inflammation during the study period. This stable state is a prerequisite for avoiding confounding factors that obscure the in vivo effects of albumin on blood immune cells.

[0219] The primary findings of the Discovery study were provided by a comparison of blood transcriptomes at T2 versus T1 in 30 patients who received albumin and 19 who did not. The inventors observed that patients treated with albumin exhibited genetic signatures not observed in those who did not receive the treatment. Therefore, patients treated with albumin were characterized by increased signatures associated with adaptive immune cells (including those associated with B cells, plasma cells, and immunoglobulins, and CD4 T cells) and signatures associated with innate myeloid mononuclear cells. Furthermore, patients treated with albumin showed increased signatures associated with the cell cycle, mitosis, and TFs (such as c-Myc and E2F family members). Importantly, results obtained using scRNA-seq in ex vivo patient PBMCs exposed to albumin indicated that albumin itself induces changes in both adaptive and innate immune cells. These findings collectively suggest that albumin administration-triggered signaling leads to expansion of B cell and CD4 T cell compartments, while simultaneously activating some myeloid mononuclear cells.

[0220] Another key finding of this study is that albumin can activate neutrophils. In patients treated with albumin, multiple genes that serve as markers of activated low-density neutrophils were upregulated. Furthermore, in vitro experiments surprisingly demonstrated that neutrophils exposed to albumin exhibited increased degranulation and enhanced phagocytosis. Therefore, albumin can restore neutrophil-mediated defense mechanisms, which are known to be severely impaired in acute decompensated cirrhosis. 5,28

[0221] Patients with acute decompensated cirrhosis have a very high prevalence of bacterial infection upon admission, reaching 37.3% among patients with ACLF. 11And the rate was 25.1% among patients who did not have ACLF. 3,4 Among patients who were not infected upon admission, 46% exhibited ACLF symptoms. 11 and 18% of patients who did not show ACLF 4 Bacterial infections occurred during hospitalization. These data suggest that the body's defense mechanisms against microbes are severely compromised. The presence of gut bacterial translocation, coupled with severe impairment of innate immune cells (as evidenced by the defective antimicrobial function of neutrophils and monocytes) and depletion of lymphocyte compartments (including T cells, B cells, and NK cells), may explain such a high risk of infection in patients with acute decompensated cirrhosis. 29 The results of this study strongly suggest that albumin can salvage the deficient antimicrobial function of neutrophils and depleted lymphocyte compartments in the most severe forms of acute decompensated cirrhosis.

[0222] An interesting observation from the in vitro study presented in this article is that the response to albumin in lymphocytes (scRNA-seq experiment) and the response in neutrophils (experiment assessing degranulation and phagocytosis) occurred in less than 2 hours, indicating that the signaling from albumin molecules is transmitted very rapidly in target cells.

[0223] In summary, the inventors have advantageously demonstrated in patients with severe acute decompensated cirrhosis that albumin promotes the expansion of B-cell and CD4 T-cell compartments, acts on monocytes and myeloid cells, and resets neutrophil antimicrobial function to normal. Therefore, it is advantageous that administering human albumin to patients with decompensated cirrhosis can achieve clinical benefits such as prevention of infection and inhibition of PAMP-induced systemic inflammation in these patients.

[0224] abbreviation

[0225] Additional materials and methods

[0226] Neutrophils were isolated from peripheral venous blood (20 ml) obtained from six patients with acute decompensated cirrhosis (AD) and five age-matched healthy subjects (HS) using the Ficoll-Hypaque method. In short, patients with AD were admitted to the liver intensive care unit of a hospital outpatient department (Barcelona, ​​Spain). Blood was obtained from four healthy donors through an agreement with the hospital outpatient blood bank. Peripheral venous blood (8 ml) was obtained by venipuncture and collected in EDTA-coated sterile, pyrogen-free tubes (Becton Dickinson, Grenoble, France). The blood sample was centrifuged at 200 g for 10 min to collect plasma. The precipitated cells were diluted to a volume of 20 ml with DPBS- / -. The diluted blood was separated into layers on 13.3 ml of Ficoll-Hypaque and centrifuged at 500 g for 25 min with the break-off. After separating the PBMC layer from the supernatant, neutrophils were collected from the pellets and incubated with pre-warmed ammonium chloride-potassium lysis buffer at room temperature for 10 minutes to remove red blood cells, followed by centrifugation at 400 g for 5 min. The red blood cell lysis procedure was repeated twice, and the resulting pellets were washed with DPBS. The separated neutrophils were resuspended in RPMI 1640 medium containing penicillin (100 U / mL), streptomycin (100 U / mL), and L-glutamine (4 mM) and free of fetal bovine serum (FBS) for the following neutrophil function assays.

[0227] threshing test After standing for 30 minutes, neutrophils increased at a rate of 3 x 10⁻⁶. 6 Seeds were inoculated at a density of 100 cells / mL and incubated with albumin, recombinant human albumin (15 mg / mL, both 15 mg / mL) or a media control in a 5% CO2 incubator at 37°C for 2 hours in the absence or presence of phorbol 12-myristate 13-acetate (100 nM). At the end of the incubation period, the supernatant was collected and threshing was measured using a neutrophil myeloperoxidase activity assay kit (Cayman Chemical, AnnArbor, MI). Briefly, 25... 1 neutrophil supernatant and 25 The assay buffer was added to each well of the experimental plate. Then, 50 μl of 3,3',5,5'-tetramethylbenzidine (TMB) (a substrate for horseradish peroxidase) was added to each well, and the absorbance was measured at 1 minute and 5 minutes after the addition of TMB using a microplate reader (Infinite M PLEX Monochromator, TECAN, Männedorf, Switzerland). The assays were performed at room temperature.

[0228] Phagocytosis assay After standing for 30 minutes, neutrophils were cultured at a rate of 5 x 10⁻⁶ cells / mL. 5 Cells were seeded at a density of [number] cells / mL and incubated for 2 hours at 37°C in a 5% CO2 incubator with albumin, recombinant human albumin (both 15 mg / mL), or a media control. Subsequently, 50 µL of conditioned fluorescein-bound yeast glycan bioparticles (Thermo Fisher Scientific) were added to each well (cell / bioparticle ratio 1:10), resulting in a final volume of 200 µL, and incubated at 37°C for 60 min. Cells were then washed with sterile DPBS and 100 µL of trypan blue solution (1 / 10 dilution in sterile DPBS) was added to quench the fluorescence of the extracellular bioparticles. Finally, the plate was centrifuged at 400 g for 5 min at room temperature, and excess trypan blue was carefully aspirated. The fluorescence intensity of each well was read using a microplate reader (FLUOstar Optima, Ortenberg, Germany).

[0229] The results showed that albumin could salvage the antimicrobial function of defective neutrophils. Therefore, it is advantageous that administering human albumin to patients with decompensated cirrhosis can achieve clinical benefits such as prevention of infection and inhibition of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in patients with decompensated cirrhosis.

[0230] Readers are advised to note all papers and documents related to this application, submitted concurrently with or prior to this specification, and made publicly available together with this specification, the contents of which are incorporated herein by reference.

[0231] All features disclosed in this specification (including any appended claims, abstract, and drawings) and / or all steps of any disclosed method or process may be combined in any combination, except that at least some of such features and / or steps are mutually exclusive combinations.

[0232] Each feature disclosed in this specification (including any appended claims, abstract, and drawings) may be replaced by an alternative feature serving the same, equivalent, or similar purpose, unless otherwise expressly stated. Therefore, unless otherwise expressly stated, each disclosed feature is merely one example of a general series of equivalent or similar features.

[0233] This invention is not limited to the details of any of the foregoing embodiments. This invention covers any novel one or any novel combination of features disclosed in this specification (including any appended claims, abstract, and drawings), or any novel one or any novel combination of steps of any disclosed method or process.

[0234] References

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[0270] Numbering Clauses

[0271] Clause 1. A composition comprising human albumin for use in treating defective immune cell function in a subject suffering from systemic inflammatory response syndrome, wherein: (a) The composition comprising human albumin is used to treat defective neutrophil function, wherein the human albumin is administered to the subject at a dose sufficient to increase the antimicrobial function of neutrophils; and / or (b) The composition comprising human albumin is used to treat defective CD4+ T cell function, wherein human albumin is administered to the patient at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells; and / or (c) The composition comprising human albumin is used to treat defective dendritic cell function, wherein human albumin is administered to the subject at a dose sufficient to increase the level of plasmacytoid dendritic cells; and / or (d) The composition comprising human albumin is used to treat defective monocyte function, wherein human albumin is administered to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

[0272] Clause 2. The composition used according to Clause 1(a), wherein the increase in neutrophil antimicrobial function is an increase in neutrophil degranulation and / or neutrophil phagocytosis.

[0273] Clause 3. The composition used according to Clause 1(a) or Clause 2, wherein human albumin is administered to a subject in a dose sufficient to upregulate the expression of neutrophil genes selected from: CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1, or combinations thereof.

[0274] Clause 4. The composition used according to any of the preceding clauses, wherein the systemic inflammatory response syndrome is associated with a disease selected from the following: decompensated cirrhosis, acute liver failure, pre-acute chronic-on-chronic liver failure, acute chronic-on-chronic liver failure, severe sepsis, septic shock, severe multiple traumatic injury, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

[0275] Clause 5. The composition used according to any of the preceding clauses, wherein the subject is a patient with sepsis or a patient with liver disease.

[0276] Clause 6. The composition used according to any of the preceding clauses, wherein the subject has a liver condition selected from the following: decompensated cirrhosis, pre-acute chronic liver failure, and acute chronic liver failure.

[0277] Clause 7. A composition comprising human albumin for use in treating defective immune cell function in a subject in need, wherein the defective immune cell function is selected from: (a) Defective neutrophil function; (b) Defective central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function; (c) Defective plasmacytoid dendritic cell function; and (d) Defective intermediate HAVCR2+ monocyte function; The defective immune cell function mentioned above is associated with diseases selected from the following: systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute chronic liver failure, chronic-on-acute liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

[0278] Clause 8. The composition for use according to any of the preceding clauses, wherein human albumin is used for administration via a route selected from: intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

[0279] Clause 9. The composition used according to any of the preceding clauses, wherein human albumin is used as part of a multi-dose regimen administered at multiple intervals.

[0280] Clause 10. The composition used according to Clause 9, wherein: The multiple-dose regimen comprises multiple fractional doses administered at equal intervals of approximately 1 day to approximately 30 days; or The multiple-dose regimen comprises multiple fractional doses administered at unequal intervals of approximately 1 day to approximately 30 days; or The multiple dosing regimen comprises two or more administrations until the total cumulative dose is reached.

[0281] Clause 11. The composition used according to any one of Clauses 9 to 10, wherein: Human albumin is used as part of a multiple-dose regimen, administered as a single dose of approximately 5 g / interval to approximately 500 g / interval; or Human albumin is used as part of a multi-dose regimen, administered in single doses of approximately 20 g / interval to approximately 200 g / interval.

[0282] Clause 12. The composition used according to any one of Clauses 9 to 11, wherein the dosing interval is every 15 days or less.

[0283] Clause 13. The composition used in accordance with any of the preceding clauses, wherein the human albumin is human plasma-derived albumin or recombinant human albumin.

[0284] Clause 14. The composition used according to any of the preceding clauses, wherein the concentration of human albumin is 4% to 25% (w / v); or wherein the concentration of human albumin is about 20% (w / v).

Claims

1. A composition comprising human albumin for use in modulating immune cells in a subject suffering from systemic inflammatory response syndrome, wherein: (a) The composition comprising human albumin is used to modulate neutrophil function, wherein the human albumin is administered to the subject at a dose sufficient to increase the antimicrobial function of neutrophils; and / or (b) The composition comprising human albumin is used to modulate CD4+ T cells, wherein the human albumin is administered to a patient at a dose sufficient to increase the level of central memory ITGB1+ CD4+ T cells and / or decrease the level of activated CD4+ memory T cells; and / or (c) The composition comprising human albumin is used to modulate dendritic cells, wherein the human albumin is administered to the subject at a dose sufficient to increase the level of plasma cell-like dendritic cells; and / or (d) The composition comprising human albumin is used to regulate monocytes, wherein the human albumin is administered to the subject at a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

2. The composition for use according to claim 1(a), wherein, Increased antimicrobial function of neutrophils is due to increased neutrophil degranulation and / or neutrophil phagocytosis.

3. The composition for use according to claim 1(a) or claim 2, wherein, The human albumin is administered to the subject in a dose sufficient to upregulate the expression of neutrophil genes selected from: CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1, or combinations thereof.

4. The composition for use according to any one of the preceding claims, wherein, The systemic inflammatory response syndrome is associated with a selection of diseases including: decompensated cirrhosis, acute liver failure, pre-acute chronic-on-chronic liver failure, acute chronic-on-chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

5. The composition for use according to any one of the preceding claims, wherein, The subjects were patients with sepsis or liver disease.

6. The composition for use according to any one of the preceding claims, wherein, The subjects had liver conditions selected from the following: decompensated cirrhosis, pre-acute chronic liver failure, and acute chronic liver failure.

7. A composition comprising human albumin for treating defective immune cell function in subjects in need, wherein, The deficient immune cell functions are selected from: (a) Defective neutrophil function; (b) Defective central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function; (c) Defective plasmacytoid dendritic cell function; and (d) Defective intermediate HAVCR2+ monocyte function.

8. The composition for use according to claim 7, wherein, The defective immune cell function is associated with a selection of diseases including: systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute chronic liver failure, acute chronic liver failure, severe sepsis, septic shock, severe multiple trauma, severe burns, acute pancreatitis, severe acute pancreatitis, hemophagocytic syndrome, heat shock syndrome, acute ischemia-reperfusion syndrome, inflammatory diseases, and cancer.

9. The composition for use according to any one of the preceding claims, wherein, The human albumin is intended for administration via a route selected from the following: intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

10. The composition for use according to any one of the preceding claims, wherein, The human albumin is used as part of a multi-dose regimen to be administered at multiple intervals.

11. The composition for use according to claim 10, wherein: Multiple-dose regimens consist of multiple partial doses administered at equal intervals of approximately 1 day to approximately 30 days; or Multiple-dose regimens consist of multiple fractional doses administered at unequal intervals of approximately 1 day to approximately 30 days; or Multiple dosing regimens involve two or more doses until the total cumulative dose is reached.

12. The composition for use according to any one of claims 10 to 11, wherein: The human albumin is used as part of a multi-dose regimen to be administered in individual doses of about 5 g / interval to about 500 g / interval; or The human albumin is used as part of a multi-dose regimen to be administered in individual doses of about 20 g / interval to about 200 g / interval.

13. The composition for use according to any one of claims 10 to 12, wherein, The dosing interval is every 15 days or less.

14. The composition for use according to any one of the preceding claims, wherein, The human albumin is either human plasma-derived albumin or recombinant human albumin.

15. The composition for use according to any one of the preceding claims, wherein, The concentration of the human albumin is 4% to 25% (w / v); or the concentration of the human albumin is about 20% (w / v).

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