Use of albumin for regulating immune cells

Abstract

The present invention relates to the use of albumin for regulating immune cells in subjects requiring it. In particular, the present invention relates to compositions comprising human albumin for use in regulating immune cells in subjects having systemic inflammatory response syndromes.

Description

[Technical Field]

[0001] The present invention relates to the use of albumin in the regulation of immune cells in subjects requiring it. In particular, the present invention relates to compositions comprising human albumin for use in the regulation of immune cells in subjects having systemic inflammatory response syndromes. [Background technology]

[0002] The immune system is a collection of cells and molecules that work together to protect humans from infectious pathogens. The immune system also provides humans with a surveillance mechanism that continuously monitors the integrity of cells and tissues. This surveillance recognizes changes in major histocompatibility complex proteins on cell membranes and viral proteins that indicate cancer development or intracellular viral infection. The immune system functions on two main principles: (1) recognition of foreign (non-self) substances by highly sensitive cell membrane or intracellular receptors present on immune cells, which leads to the expression of hundreds of genes that regulate inflammatory responses and the proliferation of immune cells; (2) elimination of microorganisms by diverse mechanisms including nonspecific antimicrobial proteins synthesized by the liver (i.e., a complement system that kills bacteria by direct lysis or activation of phagocytosis by immune cells); release of intracellular cytotoxic granules containing enzymes and pore-forming proteins (perforins) capable of killing microorganisms by granulocytes, T-cytotoxic lymphocytes (Tc cells), and natural killer (NK) cells, phagocytosis (granulocytes and macrophages), and specific antibodies [B lymphocytes (B cells)]. Activated Tc cells and NK cells present Fas ligands and interact with Fas receptors on target cells (cancer cells or virus-infected cells) to induce apoptosis-induced cell death. Antibodies, complement, and granulocytes are responsible for defense against most extracellular microorganisms, while macrophages, Tc cells, and NK cells are involved in surveillance processes and the elimination of tumor cells and virus-infected cells. Each part of the immune system functions complementaryly.

[0003] The innate immune system functions as a rapid response force, deploying a nonspecific (but highly effective) arsenal of weapons within minutes to eradicate or contain infectious pathogens. Cells of the innate immune system use conserved nonspecific pattern recognition receptors (PRRs) to reliably recognize common pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) released upon tissue injury. The recognition of PAMPs and DAMPs and the primary immune response are simple processes that occur within minutes. The innate immune system is composed of granulocytes (neutrophils, basophils, and eosinophils), macrophages (tissue cells derived from circulating monocytes), and dendritic cells (specialized cells that present antigens to T cells, B cells, and NK cells for the immune response). Although NK cells are lymphocytes, they are also part of the innate immune system.

[0004] The association of systemic inflammation, immunosuppression, a high incidence of secondary bacterial infections, and multiple organ failure identifies a syndrome commonly observed in patients with critical conditions, including acute decompensated cirrhosis, acute liver failure, severe sepsis (including septic shock), acute pancreatitis and other intraperitoneal inflammatory processes, multiple trauma, or severe burns.

[0005] Acute decompensated cirrhosis is defined as a condition in which ascites, hepatic encephalopathy, gastrointestinal bleeding, or a combination thereof has recently developed. 1、2 Acute exacerbation of chronic liver failure (ACLF) 1、2 And, Pre-ACLF 3、4 There are two severe forms. ACLF is defined as a patient presenting with single or multiple organ failure. 1、2Among patients with acute decompensated cirrhosis, immune activation is more prominent in patients with acute-on-chronic liver failure (ACLF), which is a syndrome characterized by intense systemic inflammation, multiple organ dysfunction / failure, and a high frequency 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), and this tendency is more prominent compared to non-ACLF patients (Weiss et al., 2021, Frontiers in Immunol, vol. 12, art. 699563). Pre-ACLF refers to a group of patients who are admitted with acute decompensated cirrhosis but do not have ACLF, and is defined as developing ACLF during hospitalization or within 3 months after hospitalization. 3、4 Recently, an unpublished survey has shown that pre-ACLF consists of two different phenotypes: "early" pre-ACLF, which defines patients who develop ACLF during the first hospitalization, and "late-onset" pre-ACLF, which defines patients who develop ACLF from discharge after the first hospitalization to the end of the 3-month follow-up period. In studies of ACLF patients, these patients exhibit intense systemic inflammation (indicated by leukocytosis 1、2 , neutrophilia 5 , elevated cytokine blood levels 6 , bioactive lipids 7 , and C-reactive protein 1、2 ), immunosuppression (reduced response of specific monocyte subsets to bacterial products 8~10 ; reduced microbial killing ability of neutrophils 5 , lymphopenia including T cells, B cells, and NK cells 5 ), and a high incidence of secondary infections 11 , which are fatal complications 11、12 . Patients admitted with "early" pre-ACLF also show intense systemic inflammation, but this feature is moderate in "slowly progressive" pre-ACLF patients. To date, there is no treatment for the abnormal regulation of immune cells in these patients.

[0006] Albumin is used in patients who have undergone treatment with large-volume aspiration. 13 Spontaneous bacterial peritonitis (SBP) 14 , or hepatorenal syndrome - acute kidney injury (HRS-AKI) 15 It is commonly used in patients with cirrhosis, including those with [specific condition]. The effects of albumin are usually attributed to plasma volume dilation. 11 However, recent studies suggest that albumin may affect certain immune cell functions. 16、17 This finding may explain the lower incidence of SBP and other infections reported in patients receiving long-term albumin therapy compared to patients receiving standard medical therapy. 18 .

[0007] Furthermore, although there is currently no established treatment for immunosuppression, recent studies suggest that intravenous albumin may reduce the severity of systemic inflammation beyond its plasma dilation properties (Fernandez et al., 2018, Gut, 67:1870-1880; Casulleras et al., 2020, Sci Transl Med, 12(566):eaax5135). [Overview of the Initiative] [Problems that the invention aims to solve]

[0008] The effects of intravenous albumin on lymphopenia and impaired neutrophil antimicrobial function, which are characteristic of patients with acute exacerbations of chronic liver failure (ACLF), remain unclear. [Means for solving the problem]

[0009] The present invention is based on the surprising discovery that treatment with human albumin affects different subsets of immune cells within the target (specifically neutrophils, CD4+ T cells, dendritic cells and / or monocytes). Accordingly, compositions containing human albumin are useful in modulating immune cells (specifically neutrophils, CD4+ T cells, dendritic cells and / or monocytes) in a subject in need thereof. The present invention is particularly useful in the context of subjects in which immune cells (specifically neutrophils, CD4+ T cells, dendritic cells and / or monocytes) are impaired and / or subjects who would benefit from an increase in immune cell function (specifically neutrophil function, CD4+ T cell function, dendritic cell function and / or monocyte function), for example to prevent microbial infection.

[0010] As described in the examples included herein, the inventors performed whole blood RNA sequencing (RNA-seq) at admission and 15 days (median) after the onset of ACLF in 49 inpatients with severe acute decompensated cirrhosis without ACLF. These patients were selected because of the stable course of systemic inflammation. Thirty patients received albumin administration during the progression of ACLF, while the remaining 19 did not. Additionally, in additional patients presenting with acute decompensated cirrhosis, the inventors performed single cell RNA-seq (scRNA-seq) in peripheral blood mononuclear cells (PBMCs) ex vivo exposed to albumin or vehicle for 2 hours, and also evaluated the antibacterial capacity of neutrophils ex vivo exposed to albumin.

[0011] Analysis of whole blood RNA-seq data revealed that patients receiving albumin administration specifically showed upregulation of signatures related to B cells, plasma cells, immunoglobulins; CD4 T cells; myeloid cells; mismatch repair, cell cycle and mitosis; and transcription factors such as the c-Myc and E2F families. Analyzing PBMCs from patients ex vivo exposed to albumin using scRNA-seq showed an increase in signatures related to B cells, myeloid cells, and CD4 T cells. Furthermore, the inventors demonstrated that neutrophils ex vivo exposed to albumin showed an increased degranulation response and enhanced phagocytosis.

[0012] Thus, advantageously, the inventors showed that in patients with severe acute decompensated cirrhosis, albumin also promotes the expansion of the B cell compartment and the CD4 T cell compartment, acts on mononuclear myeloid cells, and normally resets the antibacterial function of neutrophils.

[0013] Thus, a composition comprising human albumin for use in modulating immune cells in a subject having a systemic inflammatory response syndrome, (a) the composition comprising human albumin is for modulating neutrophil function, and the human albumin is for administration to the subject at a dosage sufficient to increase the antibacterial function of neutrophils; and / or (b) the composition comprising human albumin is for modulating CD4+ T cells, and the human albumin is for administration to the patient at a dosage 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 for modulating dendritic cells, and the human albumin is for administration to the subject at a dosage sufficient to increase the level of plasmacytoid dendritic cells; and / or (d) Compositions containing human albumin are intended to modulate monocyte cells, and human albumin is intended to be administered to subjects in doses sufficient to increase the level of intermediate HAVCR2+ monocytes. Compositions are provided herein.

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

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

[0016] Preferably, systemic inflammatory response syndrome may relate to diseases selected from the group consisting of decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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] Preferably, the subjects may be patients with sepsis or hepatic conditions.

[0018] Preferably, the subjects may have liver conditions selected from the group consisting of decompensated cirrhosis, the pre-stage of acute exacerbation of chronic liver failure, and acute exacerbation of chronic liver failure.

[0019] A composition comprising human albumin for treating deficient immune cell function in a subject requiring it, wherein the deficient immune cell function is (a) Deficient neutrophil function; (b) Deficient central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function; (c) Deficient plasmacytoid dendritic cell function; and (d) Deficient intermediate HAVCR2+ monocyte function A composition selected from the group consisting of the following is also provided.

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

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

[0022] Preferably, the deficient immune cell function may relate to a disease selected from the group consisting of systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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] Preferably, human albumin may be intended for administration via a route of administration selected from the group consisting of intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

[0024] Preferably, human albumin may be administered at multiple intervals as part of a multi-dose regimen.

[0025] Preferably, a multi-dose regimen may include multiple divided doses administered at equal intervals of approximately 1 to 30 days; or a multi-dose regimen may include multiple divided doses administered at unequal intervals of approximately 1 to 30 days; or a multi-dose regimen may include two or more doses until a total cumulative dose is reached.

[0026] Preferably, human albumin may be administered as part of a multi-dose regimen in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval; or human albumin may be administered as part of a multi-dose regimen in individual doses ranging from approximately 20 g / interval to approximately 200 g / interval.

[0027] Preferably, the administration interval may be every 15 days or less.

[0028] Preferably, the human albumin may be human plasma-derived albumin or recombinant human albumin.

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

[0030] As described herein, human albumin affects different subsets of immune cells within a subject (specifically neutrophils, CD4+ T cells, dendritic cells, and / or monocytes). Therefore, it is a therapeutic agent in the compositions described herein. Accordingly, the term "human albumin" may be used interchangeably with the term "composition comprising human albumin" throughout this specification.

[0031] Throughout this specification and in the claims, the words “contains” and “includes” and their variations mean “contains, but not limited to,” and are not intended (and will not) exclude other parts, additives, ingredients, integers, or steps.

[0032] Throughout this specification and in its claims, unless otherwise specified in the context, the singular form includes the plural form. In particular, where the indefinite article is used, this specification should be understood as both plural and singular unless otherwise specified in the context.

[0033] Features, integers, properties, compounds, chemical parts, or groups described in relation to specific aspects, embodiments, or examples of the present invention should be understood to be applicable to any other aspects, embodiments, or examples described herein, unless otherwise incompatible.

[0034] Various aspects of the present invention are described in further detail below.

[0035] Embodiments of the present invention are further described below with reference to the accompanying drawings. [Brief explanation of the drawing]

[0036] [Figure 1A]Whole blood gene signatures associated with albumin therapy. (A) The Euler plot shows differentially expressed genes (DEGs) between time 2 (T2) and time 1 (T1) in the albumin and non-albumin groups. DEGs were defined by an absolute fold change (FC) greater than 1.5 and P < 0.05. (B) The volcano plot plots the effect size (log2FC) of differential expression between T2 and T1 against significance (-log10 P) for the albumin (top) and non-albumin (bottom) groups. In both volcano plots, gray dots indicate genes with 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). Grayscale dots indicate DEGs, which are upregulated or downregulated genes. (C) Comparison of differential expression effect size (log2FC) between T2 and T1 in the albumin group and the effect size of the difference in the corresponding non-albumin group. Only genes included in the Gene Ontology gene set "GOCC_Immunoglobulin Complex" are shown here. DEGs with matching signs were considered to be shared. (D) Violin plots of RNA-seq estimated signatures of plasmablasts, NK cells, and T cells at T1 and T2 time points, determined using SingleR R software. Baseline values ​​in healthy subjects are also shown. White diamond symbols indicate medians. P values ​​are calculated by the Mann-Whitney U test following the Kruskal-Wallis test. (E) Heatmap of 32 differential expression (DE) blood transcription modules (BTMs) between T2 and T1 that were specific to either the albumin or non-albumin group. Thirty-one DE BTMs were specific to the albumin group, while only one DE BTM ("endoplasmic reticulum (M37.2)") was specific to the non-albumin group. The BTMs were hierarchically clustered based on the QuSAGE activity scores obtained in the albumin group. Asterisks indicate P<0.05. Grayscale represents the QuSAGE activity score (-0.4 to 0.4). [Figure 1B-1] Same as above [Figure 1B-2] Same as above [Figure 1C]Same as above [Figure 1D] Same as above [Figure 1E] Same as above [Figure 2A] Single-cell RNA sequencing identifies specific immune cell changes in peripheral blood mononuclear cells (PBMCs) ex vivo exposed to albumin. This figure is based on results obtained from PBMCs of patients with acute decompensated cirrhosis. (A) Uniform manifold approximation and projection (UMAP) of B lymphocytes from 1946 patients exposed to albumin and the media, color-coded by cell type. (B) Overlay of albumin exposure and media exposure in B lymphocyte UMAP. (C) Box plot showing the abundance of B lymphocyte populations that changed significantly after albumin exposure. (D) UMAP of myeloid cells from 21819 patients exposed to albumin and the media, displayed as cell populations. (E) Overlay of albumin exposure and media exposure in myeloid cell UMAP. (F) Box plot showing the abundance of myeloid cell type (plasmacytoid dendritic cells; pDCs) that changed significantly after albumin exposure. (G) UMAP of CD4 T cells from 12,692 patients exposed to albumin and media, color-coded by cell type. (H) Overlay of albumin exposure and media exposure in CD4 T cell UMAP. (I) Box plot showing the abundance of CD4 T cells that significantly changed after albumin exposure. (J) Left: Representative density plot showing the signature score of BTM "Mitotic Cell Cycle in Stimulated CD4 T Cells (M4.11)" in CD4 T cell UMAP under media and albumin conditions. Right: Comparison of the log (signature score) of BTM4.11 in the CD4 T cell compartment between albumin and media. Statistical analysis of BTM signatures was performed using the Wilcoxon signed-rank test. Significant p-values ​​are shown. All comparisons of cell type abundances between albumin and media were tested for significance using the paired Wilcoxon signed-rank test. Significant adjusted p-values ​​are shown. [Figure 2B] Same as above [Figure 2C] Same as above [Figure 2D] Same as above [Figure 2E] Same as above [Figure 2F] Same as above [Figure 2G] Same as above [Figure 2H] Same as above [Figure 2I] Same as above [Figure 2J-1] Same as above [Figure 2J-2] Same as above [Figure 3] The effect of albumin on the host defense function of neutrophils derived from patients with acute decompensated cirrhosis. Panels A and B were prepared using functional assays on peripheral neutrophils newly isolated from 6 patients with acute decompensated cirrhosis and 5 age-matched healthy controls. (A) Neutrophils from both patients with acute decompensated cirrhosis and healthy controls were incubated with cell culture medium, human serum albumin (15 mg / ml), or recombinant human albumin (15 mg / ml) for 2 hours in a 37°C, 5% CO2 incubator. Neutrophil degranulation was assessed by measuring MPO enzyme activity in the cell supernatant. (B) Phagocytic activity was assessed by incubating neutrophils from both patients with acute decompensated cirrhosis and healthy controls with FITC-labeled zymosan bioparticles alone, or in the presence of human serum albumin and recombinant albumin (15 mg / mL) for 60 minutes, and compared to the medium control. [Modes for carrying out the invention]

[0037] The patent documents and scientific and technical literature referenced herein establish the knowledge available to those skilled in the art at the time of filing. All disclosures of issued patents, published and pending patent applications, and other publications cited herein are incorporated herein by reference to the same extent as they would be specifically and individually indicated. In the event of any conflict, the disclosures herein shall prevail.

[0038] Various aspects of the present invention are described in further detail below.

[0039] This invention is based on the surprising discovery that treatment with human albumin affects different subsets of immune cells within a subject (specifically neutrophils, CD4+ T cells, dendritic cells, and / or monocytes).

[0040] The immune cell gene signatures of patients with acute decompensated cirrhosis (pre-ACLF) who developed ACLF after albumin treatment ("albumin group") were compared to those of equivalent patients who did not receive albumin ("non-albumin group"). A surprising number of similarities were identified in the transcriptional characteristics of pre-ACLF and ACLF patients. In addition, differential expression of multiple gene modules was observed in the albumin group. Specifically, several genes that marker activated low-density neutrophils were upregulated in albumin-treated patients. Furthermore, a significant increase in the abundance of intermediate monocytes (HAVCR2+) and plasmacytoid DCs was observed in the albumin group (Figures 2E and 2F). Finally, albumin was shown to alter the profile of the CD4+ T cell compartment (Figure 2H), with a significant decrease in the abundance of activated memory CD4+ T cells and an increase in central memory ITGB1+ CD4+ T cells (Figure 2I).

[0041] The incidence of bacterial infections upon hospitalization is very high in patients with acute decompensated cirrhosis. Among patients who were infection-free upon admission, a significant proportion develop bacterial infections during hospitalization due to severely impaired defense mechanisms against microorganisms. This study demonstrates that albumin can restore both the deficient antimicrobial function of neutrophils and the depleted lymphocyte compartment in the most severe forms of acute decompensated cirrhosis.

[0042] Neutrophil regulation The inventors hereby demonstrate that treatment with human albumin specifically upregulates nine major neutrophil genes (CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1) in subjects (Figure 1B). In addition, neutrophils ex vivo exposed to human albumin surprisingly show increased antimicrobial activity, including increased degranulation response and enhanced phagocytosis (Figure 3). The data provided herein demonstrate that human albumin can modulate neutrophil function (particularly the enhancement of neutrophil antimicrobial function) in subjects.

[0043] Accordingly, compositions comprising human albumin for use in modulating neutrophil function in a subject are provided herein, wherein the human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or increased neutrophil phagocytosis). Such compositions may be used for the prevention of infection and / or the suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in a subject. Accordingly, compositions comprising human albumin for the prevention of infection and / or the suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in a subject are also provided herein, wherein the human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or increased neutrophil phagocytosis).

[0044] In one example, a composition comprising human albumin for the prevention of microbial infection and / or suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in a subject is also provided herein, wherein the human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (for example, the increase in antimicrobial function of neutrophils is an increase in neutrophil degranulation and / or neutrophil phagocytosis).

[0045] Suitable examples of 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 increasingly involved in hospital-acquired infections. Other most common infections include urinary tract infections (UTIs), pneumonia, primary / spontaneous bloodstream infections (BSIs), and skin and soft tissue infections. The most frequently involved pathogens in UTIs are Enterobacteriaceae and enterococci, while the most frequently involved pathogens in primary BSIs are Enterobacteriaceae and staphylococci. Other relatively less common infections include cholangitis, endocarditis, catheter-related bloodstream infections, Clostridium difficile colitis, 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, Pages S101-S117. These suitable examples of microbial infections apply to all relevant embodiments described herein.

[0046] The compositions provided herein are particularly useful when administered to subjects with systemic inflammatory response syndrome (SIRS). Subjects with SIRS typically have deficient neutrophil antibacterial function. Advantageously, the compositions provided herein can be used to increase the antibacterial function of neutrophils in these subjects (e.g., increased antibacterial function of neutrophils is increased neutrophil degranulation and / or neutrophil phagocytosis), and thus can be used particularly for the prevention of infection and / or suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects with SIRS. Accordingly, compositions comprising human albumin for use in modulating neutrophil function in subjects with SIRS are provided herein, wherein the human albumin is administered to the subject in a dose sufficient to increase the antibacterial function of neutrophils (e.g., increased antibacterial function of neutrophils is increased neutrophil degranulation and / or neutrophil phagocytosis).

[0047] Therefore, compositions provided herein may be intended for the treatment of SIRS in subjects by modulating neutrophil function. Compositions comprising human albumin for the prevention of infection and / or suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects having SIRS are also provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (for example, the increase in antimicrobial function of neutrophils is an increase in neutrophil degranulation and / or neutrophil phagocytosis). Therefore, compositions provided herein may be intended for the treatment of SIRS in subjects by the prevention of infection and / or suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation.

[0048] Examples of subjects with SIRS are described elsewhere in this specification. For example, SIRS may relate to diseases selected from the group consisting of decompensated cirrhosis, acute liver failure, the pre-acute exacerbation of chronic liver failure, acute exacerbation of 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. In other words, subjects with SIRS may also have diseases selected from the group listed above.

[0049] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS may 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.

[0050] Compositions comprising human albumin for use in modulating neutrophil function in subjects having decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or neutrophil phagocytosis). Thus, compositions provided herein may be intended for treating decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by modulating neutrophil function. Furthermore, compositions comprising human albumin for the prevention of infection and / or suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects having decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or increased neutrophil phagocytosis). Thus, compositions provided herein may be intended for the treatment of decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by preventing infection and / or suppressing pathogen-associated molecular pattern (PAMP)-induced systemic inflammation.

[0051] The compositions described herein are also useful when administered to subjects with deficient neutrophil function [for example, subjects with deficient neutrophil antibacterial function (e.g., the antibacterial function of deficient neutrophils may be degranulation and / or phagocytosis of deficient neutrophils)]. Advantageously, the compositions provided herein can be used to increase the antibacterial function of neutrophils in these subjects (e.g., an increase in the antibacterial function of neutrophils is an increase in neutrophil degranulation and / or neutrophil phagocytosis), and therefore can be used particularly in subjects with deficient neutrophil function to prevent infection and / or suppress pathogen-associated molecular pattern (PAMP)-induced systemic inflammation. Accordingly, compositions comprising human albumin for the treatment of deficient neutrophil function in subjects requiring it are provided herein. In this example, human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or neutrophil phagocytosis). Compositions comprising human albumin for the prevention of infection and / or suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation in subjects with deficient neutrophil function are also provided herein. In this example, human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or neutrophil phagocytosis).

[0052] Examples of subjects with impaired neutrophil function can be readily identified by those skilled in the art. Deficiencies in neutrophil function may manifest as deficiencies in various biological processes involved in neutrophil bactericidal activity, including deficiencies in arrest or adhesion on endothelial cells, deficiencies in oxidative bursts, deficiencies in extracellular bactericidal mechanisms such as neutrophil extracellular traps or NETS release, deficiencies in neutrophil degranulation, and / or deficiencies in neutrophil phagocytosis.

[0053] For example, neutrophil function deficiency may be associated with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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. Therefore, neutrophil function deficiency may be observed in subjects with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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. Accordingly, compositions comprising human albumin as described herein may be used to treat neutrophil function deficiencies in diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute hepatic failure, the pre-stage of acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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, these compositions may be used to treat aspects of neutrophil function deficiencies in the described diseases). Compositions comprising human albumin for the treatment of diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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 are therefore also provided herein, and such compositions are for modulating neutrophil function (for example, for treating neutrophil function that is deficient).

[0054] In one example, a subject with deficient neutrophil function may have decompensated cirrhosis. For instance, a subject with deficient neutrophil function may have 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.

[0055] In one example, a composition provided herein comprises human albumin, which is administered to a subject in a dose sufficient to upregulate the expression of a neutrophil gene, or a combination thereof, selected from the group consisting of CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1. This example applies to all appropriate embodiments described herein, including cases where the human albumin-containing composition is used to modulate immune cells in a subject (e.g., a subject with SIRS and / or decompensated cirrhosis), or where the human albumin composition is for the treatment of deficient neutrophil function in a subject requiring it.

[0056] CD177 (NCBI Gene ID: 57126; UniProtKB / Swiss-Prot: Q8N6Q3) encodes a glycosyl-phosphatidylinositol (GPI)-conjugated cell surface glycoprotein involved in neutrophil activation. This protein binds to platelet endothelial cell adhesion molecule-1 and can act on neutrophil migration. Mutations in this gene are associated with myeloproliferative disorders. Overexpression of this gene has been observed in patients with polycythemia vera. Autoantibodies against this protein may cause pulmonary transfusion reactions and may be involved in Wegener's granulomatosis.

[0057] Olfactomedin 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 and defines a subset of neutrophils ranging from 5% to 40% as OLFM4-positive. OLFM4 has been proposed to inhibit cathepsin C, a cysteine ​​protease essential for the activation of serine proteases.

[0058] The proteoglycan 2 (PRG2) gene (NCBI Gene ID: 5553; UniProtKB / Swiss-Prot: P13727, also known as proeosinophil major basic protein) encodes a major component of the crystalline core of eosinophil granules. PRG2 protein precursors are also present at high levels in the placenta and pregnancy serum, existing as complexes with several other proteins, including pregnancy-associated plasma protein A (PAPPA), angiotensinogen (AGT), and C3dg. PRG2 protein is involved in antiparasitic defense mechanisms as a cytotoxin and helminthotoxin, and may be involved in immunohypersensitivity reactions. PRG2 protein contains peptides that exhibit potent antibacterial activity against Gram-positive bacteria, Gram-negative bacteria, and fungi. It is directly involved in epithelial cell damage, shedding, and bronchospasm in allergic diseases.

[0059] Myeloperoxidase (MPO-NCBI Gene ID:4353;UniProtKB / Swiss-Prot:P05164) is a heme protein synthesized during myeloid differentiation and is a major component of neutrophil azurophilic granules. After being produced as a single-chain precursor, myeloperoxidase is cleaved into light and heavy chains. Mature myeloperoxidase is a tetramer consisting of two light chains and two heavy chains. MPO produces hypohalite, which is central to the bactericidal activity of neutrophils.

[0060] Bactericidal permeability-enhancing protein (BPI-NCBI Gene ID:671;UniProtKB / Swiss-Prot:P17213) encodes a lipopolysaccharide-binding protein. It is associated with human neutrophil granules and possesses antibacterial activity against Gram-negative bacteria. BPI is a potent antibacterial protein of neutrophils, possessing bactericidal activity and LPS neutralizing activity, and may also be involved in neutrophil-mediated phagocytosis of Gram-negative bacteria by promoting complement activation.

[0061] Resistin (RETN-NCBI Gene ID:56729;UniProtKB / Swiss-Prot:Q9HD89) belongs to a family defined by mouse resistin-like genes. A characteristic feature of this family is the C-terminal elongation of 10 equally spaced cysteine ​​residues. RETN proteins have antibacterial activity in the skin, exhibiting antibacterial activity against both Gram-positive and Gram-negative bacteria. RETN promotes pro-inflammatory activation of neutrophils and neutrophil extracellular trap formation.

[0062] Lipocalcin 2 (LCN2-NCBI Gene ID:3934;UniProtKB / Swiss-Prot:P80188) encodes a protein belonging to the lipocalin family. Members of this family transport small hydrophobic molecules such as lipids, steroid hormones, and retinoids. The LCN2 protein is a neutrophil gelatinase-associated lipocalin and plays a role in innate immunity by capturing iron-containing siderophores, thereby suppressing bacterial growth. The presence of LCN2 protein in blood and urine is an early biomarker of acute kidney injury. LCN2 is thought to be involved in multiple cellular processes, including maintaining skin homeostasis and suppressing infiltration and metastasis. Mice lacking this gene are more susceptible to bacterial infections than wild-type mice.

[0063] CEA cell adhesion molecule 8 (CEACAM8-NCBI Gene ID:1088;UniProtKB / Swiss-Prot:P31997) is expressed only in granulocytes and encodes a cell surface glycoprotein involved in cell adhesion in a calcium-independent manner. CEACAM8 mediates heterogeneous cell adhesion with other carcinoembryonic antigen-associated cell adhesion molecules such as CEACAM6. Furthermore, heterogeneous interactions with CEACAM8 also occur in activated neutrophils.

[0064] 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 pulmonary mast cells and alveolar macrophages. MCEMP1 is one of the most easily induced genes in many inflammatory diseases, including asthma, idiopathic pulmonary fibrosis, cancer, sepsis, and stroke.

[0065] As used herein, “modulation of neutrophil function” means controlling neutrophil function. Neutrophil function may be controlled by increasing or decreasing neutrophil activity (for example, by stimulating existing neutrophils or by increasing the total number of neutrophils present). In the context of the present invention, it is desirable to modulate neutrophil function by increasing the antimicrobial function of neutrophils (also referred herein as neutrophil antimicrobial activity). An increase in neutrophil antimicrobial function may also be referred to as enhancement of neutrophil antimicrobial function.

[0066] Neutrophils are known to perform multiple antimicrobial functions in vivo. These antimicrobial functions include neutrophil degranulation and neutrophil phagocytosis. As used herein, an increase in neutrophil antimicrobial function may refer to an increase in neutrophil degranulation and / or neutrophil phagocytosis. In some cases, an increase in neutrophil antimicrobial function may refer to an increase in a combination of these features, namely an increase in neutrophil degranulation and neutrophil phagocytosis.

[0067] Methods for determining whether or not there is an increase in the antibacterial function of neutrophils are well known. Examples of such methods are described in the Examples section below.

[0068] During host defense responses, neutrophils release myeloperoxidase (MPO), an essential antimicrobial protein primarily localized in azurofil granules or primary neutrophil granules. Therefore, increased neutrophil degranulation can be quantified by measuring MPO activity. This increased neutrophil degranulation may also be referred to as neutrophil degranulation enhancement in this specification.

[0069] Phagocytosis can be evaluated by measuring the uptake of fluorescently labeled zymosan particles by neutrophils. This method can be used to determine whether neutrophil phagocytosis is increased. Increased neutrophil phagocytosis may also be referred to as enhanced neutrophil phagocytosis in this specification.

[0070] Corresponding methods are also provided for regulating neutrophil function in subjects (and / or for the prevention of infection and / or the suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation), wherein a composition comprising human albumin is administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils (e.g., increased antimicrobial function of neutrophils is increased neutrophil degranulation and / or neutrophil phagocytosis). Corresponding methods are also provided for treating deficient neutrophil function in subjects requiring it.

[0071] Regulation of CD4+ T cells The inventors hereby demonstrate that treatment with albumin specifically upregulates the blood transcription module (BTM) "mitotic cell cycle (M4.11) in stimulated CD4 T cells" (Figure 1E) in subjects, indicating activation of transcription in CD4+ T cells. Furthermore, it is shown herein that albumin administration alters the profile of the CD4+ T cell compartment in subjects (Figure 2H), significantly reducing the abundance of activated memory CD4+ T cells and increasing central memory ITGB1+ CD4+ T cells (Figure 2I). The data provided herein demonstrate that albumin modulates the CD4+ T cell population in subjects (particularly by increasing the level of central memory ITGB1+ CD4+ T cells and decreasing the level of activated CD4+ memory T cells), alters transcription in CD4+ T cells, and thereby alters CD4+ T cell function.

[0072] Accordingly, compositions comprising human albumin for use in modulating CD4+ T cells in a subject are provided herein, wherein the human albumin is administered to the subject in 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 may be used to modulate the function of CD4+ T cells in a subject. Central memory ITGB1+ CD4+ T cells have recently been shown to possess cytotoxic properties (JI 2021, 207: 2966-2975), suggesting that these cells are involved in host resistance to infection. Accordingly, compositions provided herein may be used to prevent infection in a subject. Accordingly, compositions comprising human albumin for preventing infection in a subject are also provided herein, wherein the human albumin is administered to the subject in 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.

[0073] The compositions provided herein are particularly useful when administered to subjects with systemic inflammatory response syndrome (SIRS). Subjects with SIRS may have deficient CD4+ T cell function (particularly a deficiency in central memory ITGB1+ CD4+ T cell levels and / or an excess in activated CD4+ memory T cell levels). 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 used particularly to prevent infection in subjects with SIRS. Accordingly, compositions comprising human albumin for use in the regulation of CD4+ T cells (e.g., regulation of CD4+ T cell function) in subjects with SIRS are provided herein, and the human albumin is intended to be administered to the subject in 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, compositions provided herein may be intended for treating SIRS in a subject by modulating CD4+ T cells (e.g., modulating CD4+ T cell function). Accordingly, compositions comprising human albumin for preventing infection in a subject having SIRS are also provided herein, wherein the human albumin is intended to be administered to the subject in 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. Accordingly, compositions provided herein may be intended for treating SIRS in a subject by preventing infection.

[0074] Examples of objects having SIRS are described elsewhere in this specification. These examples are similarly applicable to this embodiment.

[0075] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS may 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.

[0076] Compositions comprising human albumin for use in regulating CD4+ T cells (e.g., modulating CD4+ T cell function) in subjects having decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is intended to be administered to the subject in 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. Thus, compositions provided herein may be intended for treating decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by regulating CD4+ T cells (e.g., modulating CD4+ T cell function). Furthermore, compositions comprising human albumin for preventing infection in subjects having decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis, or ACLF decompensated cirrhosis) are also provided herein, wherein the human albumin is intended to be administered to the subject in 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. Thus, compositions provided herein may be intended for treating decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis, or ACLF decompensated cirrhosis) in subjects by preventing infection.

[0077] Furthermore, the compositions provided herein are also useful when administered to subjects having deficient CD4+ T cell function (e.g., deficient 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 therefore can be used particularly to prevent infection in subjects having deficient CD4+ T cell function. Accordingly, compositions comprising human albumin for the treatment of deficient CD4+ T cell function in subjects requiring it are provided herein. In this example, the human albumin is administered to the subject in 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. Compositions comprising human albumin for preventing infection in subjects having deficient CD4+ T cell function are also provided herein. In this example, human albumin is administered to the subject in 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.

[0078] Examples of subjects with deficient CD4+ T cell function can be easily identified by those skilled in the art. For example, deficient CD4+ T cell function may be associated with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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. Therefore, deficient CD4+ T cell function may be observed in subjects with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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. Accordingly, compositions comprising human albumin as described herein may be used to treat deficient CD4+ T cell function in diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute hepatic failure, pre-acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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, these compositions may be used to treat aspects of deficient CD4+ T cell function in the diseases described herein).

[0079] Compositions comprising human albumin for the treatment of diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute hepatic failure, pre-acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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 are therefore also provided herein, and such compositions are for modulating CD4+ T cell function (for example, for treating deficient CD4+ T cell function).

[0080] In one example, a subject with deficient CD4+ T cell function is a subject with decompensated cirrhosis. For example, a subject with deficient CD4+ T cell function may 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.

[0081] As used herein, “modulation of CD4+ T cells” means controlling CD4+ T cells. This includes, for example, controlling the number of CD4+ T cells, controlling the population of CD4+ T cells, and controlling CD4+ T cell function. CD4+ T cell function may 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 increasing the (relative) abundance of the desired cells (to increase the number of cells having the desired T cell activity). Similarly, CD4+ T cell function may be controlled by decreasing CD4+ T cell activity, for example, by inactivating an undesirable subset of CD4+ T cells (to decrease the desired T cell activity of these cells) and / or decreasing the (relative) abundance of the undesirable cells (to decrease the number of cells having the undesirable T cell activity). In the context of the present invention, it is desirable to modulate 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.

[0082] As used herein, the term “level” refers to the quantity of a specified cell (e.g., central memory ITGB1+ CD4+ T cells or activated CD4+ memory T cells). A level may be an absolute quantity of cells and / or a relative quantity of a specified cell. A relative quantity of a specified cell may be a relative quantity of other cells in the subject, or other cells in a sample from the subject. The “other cells” may be, for example, all leukocytes, or a subpopulation of specific cells such as CD4+ T cells. A relative quantity may be, for example, a percentage, fraction, or ratio. In the context of relative quantity, it will be understood that a relative quantity of a specified cell may change (e.g., increase or decrease) regardless of whether it changes the absolute quantity of the specified cell. Similarly, the absolute quantity of a specified cell may change (increase or decrease) regardless of whether it changes the relative quantity.

[0083] Methods for determining whether there is an increase in the level of central memory ITGB1+ CD4+ T cells and / or a decrease in activated CD4+ memory T cells are well known. Examples of such methods are described in the Examples section below.

[0084] Methods are also provided for regulating (and / or preventing infection of) CD4+ T cell function in subjects, in which a composition containing human albumin is administered to the subject in 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. Corresponding methods are also provided for treating deficient CD4+ T cell function in subjects requiring it.

[0085] Regulation of dendritic cells The inventors hereby demonstrate that treatment with albumin specifically increases the amount of plasmacytoid dendritic cells in subjects (Figures 1E and 2D-2F). The data provided herein show that albumin can modulate dendritic cells (particularly by increasing the level of plasmacytoid dendritic cells), alter transcription in dendritic cells, and thereby alter dendritic cell function.

[0086] Accordingly, compositions comprising human albumin for use in modulating dendritic cells (DCs) in a subject are provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the levels of plasmacytoid dendritic cells (pDCs). Such compositions may be used to modulate the function of dendritic cells in a subject. Plasmacytoid DCs are DCs specialized for interferon-mediated antiviral responses. Accordingly, compositions provided herein may be used to prevent viral infection in a subject. Accordingly, compositions comprising human albumin for preventing viral infection in a subject are also provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the levels of plasmacytoid dendritic cells.

[0087] The compositions provided herein are particularly useful when administered to subjects with systemic inflammatory response syndrome (SIRS). Subjects with SIRS may have deficient dendritic cell function (particularly a deficiency in plasmacytoid dendritic cell levels). Advantageously, the compositions provided herein can be used to increase the levels of plasmacytoid dendritic cells in these subjects and, therefore, can be used particularly to prevent viral infections in subjects with SIRS. Accordingly, compositions comprising human albumin for use in regulating dendritic cells (e.g., regulating dendritic cell function) in subjects with SIRS are provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the levels of plasmacytoid dendritic cells. Thus, the compositions provided herein may be intended for treating SIRS in subjects by regulating dendritic cells (e.g., regulating dendritic cell function). Compositions comprising human albumin for preventing viral infections in subjects with SIRS are also provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the levels of plasmacytoid dendritic cells. Therefore, the compositions provided herein may be intended for treating SIRS in a subject by preventing viral infection.

[0088] Examples of objects having SIRS are described elsewhere in this specification. These examples are similarly applicable to this embodiment.

[0089] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS may 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.

[0090] Compositions comprising human albumin for use in regulating dendritic cells (e.g., regulating dendritic cell function) in subjects with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the level of plasmacytoid dendritic cells. Thus, the compositions provided herein may be intended to treat decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by regulating dendritic cells (e.g., dendritic cell function). Furthermore, compositions comprising human albumin for preventing viral infection in subjects with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis), wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the level of plasmacytoid dendritic cells. Therefore, the compositions provided herein may be intended for the treatment of decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in a subject by preventing viral infection.

[0091] The compositions provided herein are also useful when administered to subjects having deficient dendritic cell function (e.g., deficient plasmacytoid dendritic cell function). Advantageously, the compositions provided herein can be used to increase the levels of plasmacytoid dendritic cells in these subjects and, therefore, can be used particularly to prevent viral infections in subjects having deficient dendritic cell function. Accordingly, compositions comprising human albumin for the treatment of deficient dendritic cell function in subjects requiring it are provided herein. In this example, the human albumin is administered to the subject in a dose sufficient to increase the levels of plasmacytoid dendritic cells. Compositions comprising human albumin for the prevention of viral infections in subjects having deficient dendritic cell function are also provided herein. In this example, the human albumin is administered to the subject in a dose sufficient to increase the levels of plasmacytoid dendritic cells.

[0092] Examples of subjects with deficient dendritic cell function can be easily identified by those skilled in the art. For example, deficient dendritic cell function may be associated with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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. Therefore, deficient dendritic cell function may be observed in subjects with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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. Accordingly, compositions comprising human albumin as described herein may be used to treat deficient dendritic cell function in diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute hepatic failure, the pre-stage of acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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, these compositions may be used to treat aspects of deficient dendritic cell function in the diseases described herein).

[0093] Compositions comprising human albumin for the treatment of diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute hepatic failure, pre-acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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 are therefore also provided herein, and such compositions are for modulating dendritic cell function (for example, for treating deficient dendritic cell function).

[0094] In one example, a subject with deficient dendritic cell function is a subject with decompensated cirrhosis. For example, a subject with deficient dendritic cell function may 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.

[0095] As used herein, “modulation of dendritic cells” means controlling dendritic cells. This includes, for example, controlling the number of dendritic cells, controlling the population of dendritic cells, and controlling dendritic cell function. Dendritic cell function may 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 increasing the (relative) abundance of desired cells (to increase the number of cells having the desired dendritic cell activity). In the context of the present invention, it is desirable to modulate dendritic cell function by increasing the level of plasmacytoid dendritic cells.

[0096] As used herein, the term “level” refers to the quantity of a specified cell (e.g., plasma cell-like DCs). A level may be an absolute quantity of cells and / or a relative quantity of a specified cell. A relative quantity of a specified cell may be a relative quantity of other cells in the subject, or other cells in a sample from the subject. The “other cells” may be, for example, all leukocytes, or a subpopulation of specific cells such as dendritic cells. A relative quantity may be, for example, a percentage, fraction, or ratio. In the context of relative quantities, it will be understood that a relative quantity of a specified cell may change (e.g., increase or decrease) regardless of whether it changes the absolute quantity of the specified cell. Similarly, the absolute quantity of a specified cell may change (increase or decrease) regardless of whether it changes the relative quantity.

[0097] Methods for determining whether or not there is an increase in the level of plasmacytoid dendritic cells are well known. Examples of such methods are described in the Examples section below.

[0098] Plasma cell-like dendritic cells (pDCs) play a unique role in linking innate and adaptive immunity. They possess a combination of lymphocyte-like morphology and plasma cell characteristics, including extensive endoplasmic reticulum, multiple mitochondria, and a small Golgi apparatus. While pDCs have low antigen-presenting ability, particularly for exogenous antigens, they acquire antigen-presenting cell function after activation, accompanied by the expression of co-stimulatory molecules, enabling them to guide T cells to specific functional subsets. After TLR-mediated activation, the expression of co-stimulatory molecules is induced in all pDCs, thereby conferring T cell priming properties to these cells. pDCs are activated via different cell surface receptors and cytoplasmic nucleic acid sensors, but for IFN production, nucleic acid sensing via TLR7 and TLR9 is considered the primary mode of activation. Signaling via these two TLRs leads to rapid and massive production of type I and type III IFNs, inducing the induction of IFN-stimulating genes (ISGs), many of which possess antiviral properties. Based on these characteristics, it has been proposed that the primary function of pDCs is to act as antiviral cells. pDC activation and IFN-1 production are important for the antiviral response and can promote tissue repair. However, chronic or prolonged activation of these cells can cause or contribute to immune dysfunction and disease progression, as can be observed in autoimmune diseases and persistent viral infections.

[0099] Therefore, as used herein, “deficient pDC” or “deficient pDC function” may refer to pDCs that are chronically or persistently activated. Deficient pDC function also refers to pDCs with reduced production or expression of IFN molecules. In some cases, a deficient pDC may refer to a pDC that overproduces IFN molecules such as IFN-I, as in systemic lupus erythematosus or psoriasis. This may lead to abnormal T cell production. In some cases, a deficient pDC may produce immunosuppressive mediators such as OX40L or ICOSL, which at least partially inhibit other immune cells.

[0100] Markers of dysfunctional pDCs can sometimes be detected by determining the levels of cytokines such as IFN molecules. Another marker of pDC dysfunction may be elevated Tim-3 expression.

[0101] Methods are also provided for regulating dendritic cell function (and / or preventing viral infection) in subjects, in which a composition containing human albumin is administered to the subject in a dose sufficient to increase the level of plasmacytoid dendritic cells. Corresponding methods are also provided for treating deficient dendritic cell function in subjects requiring it.

[0102] Adjusting a single ball The inventors hereby demonstrate that treatment with albumin specifically increases the amount of intermediate HAVCR2+ monocytes in subjects (Figures 2D-2F). The data provided herein show that albumin can modulate monocytes (particularly by increasing the level of intermediate HAVCR2+ monocytes), alter transcription in monocytes, and thereby alter monocyte function.

[0103] Therefore, compositions comprising human albumin for use in modulating monocytes in a subject are provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Such compositions may be used to modulate the function of monocytes in a subject. Intermediate HAVCR2+ monocytes are monocytes that suppress inflammatory responses from other immune cells (e.g., classical monocytes, CD4 T cells) (Front. Immunol 2016;7, 229). HAVCR2 (also known as Tim3) is an immune checkpoint molecule. Therefore, compositions provided herein may be used to suppress inflammatory responses in a subject. Therefore, compositions comprising human albumin for suppressing inflammatory responses in a subject are also provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

[0104] The compositions provided herein are particularly useful when administered to subjects with systemic inflammatory response syndrome (SIRS). Subjects with SIRS may have deficient monocyte function (particularly a deficiency in intermediate HAVCR2+ monocyte levels). Advantageously, the compositions provided herein can be used to increase the levels of intermediate HAVCR2+ monocytes in these subjects and, therefore, to suppress the inflammatory response, particularly in subjects with SIRS. Accordingly, compositions comprising human albumin for use in monocyte regulation (e.g., regulation of monocyte function) in subjects with SIRS are provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the levels of intermediate HAVCR2+ monocytes. Thus, the compositions provided herein may be intended for treating SIRS in subjects by monocyte regulation (e.g., regulation of monocyte function). Compositions comprising human albumin for suppressing the inflammatory response in subjects with SIRS are also provided herein, wherein the human albumin is intended to be administered to the subject in a dose sufficient to increase the levels of intermediate HAVCR2+ monocytes. Therefore, the compositions provided herein may be intended for treating SIRS in a subject by suppressing the inflammatory response.

[0105] Examples of objects having SIRS are described elsewhere in this specification. These examples are similarly applicable to this embodiment.

[0106] In one example, a subject with SIRS is a subject with decompensated cirrhosis. For example, a subject with SIRS may 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.

[0107] Compositions comprising human albumin for use in monocyte regulation (e.g., regulation of monocyte function) in subjects with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) are provided herein, wherein the human albumin is administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Thus, the compositions provided herein may be for treating decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in subjects by monocyte regulation (e.g., regulation of monocyte function). Furthermore, compositions comprising human albumin for suppressing inflammatory responses in subjects with decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) are also provided herein, wherein the human albumin is administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Therefore, the compositions provided herein may be intended for the treatment of decompensated cirrhosis (e.g., pre-ACLF decompensated cirrhosis or ACLF decompensated cirrhosis) in a subject by suppressing the inflammatory response.

[0108] The compositions provided herein are also useful when administered to subjects with deficient monocyte function (e.g., deficient 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, to suppress inflammatory responses, particularly in subjects with deficient monocyte function. Accordingly, compositions comprising human albumin for the treatment of deficient monocyte function in subjects requiring it are provided herein. In this example, the human albumin is administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Furthermore, compositions comprising human albumin for suppressing inflammatory responses in subjects with deficient monocyte function are also provided herein. In this example, the human albumin is administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes.

[0109] Examples of subjects with deficient monocyte function can be easily identified by those skilled in the art. For example, deficient monocyte function may be associated with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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. Therefore, deficient monocyte function may be observed in subjects with diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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. Accordingly, compositions comprising human albumin as described herein may be used to treat monocyte function deficiencies in diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute hepatic failure, the pre-stage of acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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, these compositions may be used to treat aspects of monocyte function deficiencies in the diseases described herein).

[0110] Compositions comprising human albumin for the treatment of diseases selected from the group consisting of SIRS, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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 are therefore also provided herein, and such compositions are for modulating monocyte function (for example, for treating monocyte function that is deficient).

[0111] In one example, a subject with deficient monocyte function is a subject with decompensated cirrhosis. For example, a subject with deficient monocyte function may 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.

[0112] As used herein, “monocyte regulation” means controlling monocytes. This includes, for example, controlling the number of monocytes, controlling the monocyte population, and controlling monocyte function. Monocyte function may be controlled by increasing or decreasing monocyte activity, for example, by stimulating a desired subset of monocyte cells (to increase the desired monocyte activity of these cells) and / or increasing the (relative) abundance of desired cells (to increase the number of cells having the desired monocyte activity). In the context of the present invention, it is desirable to regulate monocyte function by increasing the level of intermediate HAVCR2+ monocytes.

[0113] As used herein, the term “level” refers to the quantity of a specified cell (e.g., intermediate HAVCR2+ monocytes). A level may be an absolute quantity of cells and / or a relative quantity of a specified cell. A relative quantity of a specified cell may be a relative quantity of other cells in the subject, or other cells in a sample from the subject. The “other cells” may be, for example, all leukocytes, or a subpopulation of specific cells such as monocytes. A relative quantity may be, for example, a percentage, fraction, or ratio. In the context of relative quantity, it will be understood that a relative quantity of a specified cell may change (e.g., increase or decrease), regardless of whether or not it changes the absolute quantity of the specified cell. Similarly, the absolute quantity of a specified cell may change (increase or decrease), regardless of whether or not it changes the relative quantity.

[0114] Methods for determining whether there is an increase in the level of intermediate HAVCR2+ monocytes are well known. Examples of such methods are described in the Examples section below.

[0115] Methods are also provided to address the modulation of monocyte function (and / or suppression of inflammatory responses) in subjects, in which a composition containing human albumin is administered to the subject in a dose sufficient to increase the level of intermediate HAVCR2+ monocytes. Corresponding methods are also provided for treating deficient monocyte function in subjects requiring it.

[0116] composition The compositions provided herein include human albumin.

[0117] Those skilled in the art will understand that the reference to “albumin” in this specification encompasses proteins having a tertiary structure identical and / or very similar to human serum albumin (HSA) or the HSA domain, and possessing similar properties to HSA or related domains. The term albumin includes variants and / or derivatives such as fusions and / or conjugations of albumin or albumin variants. The term “variant” means a polypeptide derived from parental albumin that includes changes at one or more (several) positions, i.e., substitutions, insertions, and / or deletions. Substitution means replacing an amino acid occupying a position with a different amino acid, deletion means removing an amino acid occupying a position, and insertion means adding one to three amino acids adjacent to an amino acid occupying a position. These altered polypeptides (variants) can be obtained through human intervention by modifying the polynucleotide sequence encoding naturally occurring (wild-type) albumin.

[0118] In preferred embodiments of the present invention, albumin is human albumin, preferably human albumin purified from human plasma (also referred herein as human plasma-derived albumin, human serum albumin, or serum albumin). Examples of commercially available human serum albumins include Albumein® and Plasbumin® (Grifols). In some examples, albumin is recombinant human albumin. Several examples of recombinant human albumins are known in the art and commercially available recombinant human albumins, such as Recombumin® (Albumedix), Cellastim S® (InVitria), and Albagen® (Albumin Bioscience), as well as human albumins expressed in plants such as rice and in yeasts such as Saccharomyces cerevisiae and Pichia pastoris.

[0119] As used herein, the term “recombinant” refers to a biomolecule such as a gene or protein that is (1) removed from its natural environment, (2) unrelated to all or part of the polynucleotides in which a gene is found in nature, (3) operably ligated with polynucleotides that are not ligated in nature, or (4) not present in nature. The term “recombinant” can also be used to refer to cloned DNA isolates, chemically synthesized polynucleotide analogs, or polynucleotide analogs biologically synthesized by heterologous systems, as well as proteins and / or mRNA encoded by such nucleic acids. In some embodiments, albumin is recombinant albumin, for example, recombinant human albumin.

[0120] As used herein, the term “human plasma-derived” refers to biomolecules, such as genes or proteins, obtained from standards of pooled human plasma from donors. In some embodiments, the term “human plasma-derived” is used to refer to human plasma-derived albumin.

[0121] The compositions described herein are for administration to subjects. Therefore, they may be referred to as pharmaceutical compositions. These compositions may contain additional activators (in addition to human albumin described herein, e.g., serum albumin or recombinant human albumin), provided that human albumin is present in a level (amount, concentration, or dose) sufficient to enable the described function. In other words, the compositions described herein contain a level (amount, concentration, or dose) of human albumin sufficient to modulate the immune cells of a subject in the manner described.

[0122] The pharmaceutical composition may include, along with the human albumin described herein, pharmaceutically acceptable excipients, adjuvants, diluents, and / or carriers.

[0123] The composition may typically contain salts, buffers, preservatives, suitable carriers, adjuvants, and other immunostimulants such as cytokines, as well as other therapeutic agents or compounds as appropriate, in pharmaceutically acceptable concentrations.

[0124] As used herein, “pharmaceutically acceptable” means that a substance is not biologically or otherwise undesirable; that is, when administered to an individual together with human albumin, the substance does not cause any undesirable biological effects or adverse interactions with any of the other components of the pharmaceutical composition in which it is contained.

[0125] Excipients are natural or synthetic substances formulated together with the active ingredient (e.g., human albumin provided herein) and are included for the purpose of increasing the volume of the formulation or enhancing the therapeutic effect of the active ingredient in the final dosage form, such as promoting drug absorption or solubility. Excipients are also useful in the manufacturing process, assisting in the handling of the relevant active ingredient, such as by promoting powder flowability or imparting non-stickiness, and contributing to in vitro stability, such as preventing denaturation during the expected storage period. Pharmaceutically acceptable excipients are well known in the art. Therefore, a suitable excipient is easily identifiable to those skilled in the art. Examples of suitable pharmaceutically acceptable excipients include water, physiological saline, aqueous dextrose, glycerol, and ethanol.

[0126] An adjuvant is a pharmacological and / or immunological agent that modifies the effects of other agents in a pharmaceutical formulation. Pharmaceutically acceptable adjuvants are well known in the art. Therefore, a suitable adjuvant is readily identifiable to those skilled in the art.

[0127] A diluent is a chemical used to dilute a substance. Pharmaceutically acceptable diluents are well known in the art. Therefore, a suitable diluent can be easily identified by those skilled in the art.

[0128] The carrier is non-toxic to the recipient at the dosage and concentration used and is compatible with the other components of the formulation. The term "carrier" refers to a natural or synthetic organic or inorganic component used to formulate the active ingredient and facilitate its application. Pharmaceutically acceptable carriers are well known in the art. Therefore, a suitable carrier is readily identifiable to those skilled in the art.

[0129] As used herein, “Subject” means an individual having, or at risk of having, a particular condition, disorder, or symptom, such as a human being. A Subject may be a patient, i.e., a Subject requiring treatment according to the present invention. A Subject may have previously received treatment for the condition, disorder, or symptom. Alternatively, a Subject may not have received treatment prior to treatment according to the present invention.

[0130] The compositions described herein may be administered to a subject by any conventional route, such as by injection or by slow infusion over time. For example, administration may be carried out by a route of administration selected from the group consisting of intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

[0131] The compositions described herein may be in any form suitable for the above-described administration method. Appropriate dosages of the compositions described herein may be determined by those skilled in the art.

[0132] The compositions described herein are intended to be administered in an effective dose. An "effective dose" is the amount, alone or in combination with additional doses, that produces a desired (therapeutic or non-therapeutic) response. The effective dose used depends, for example, on the therapeutic (or non-therapeutic) purpose, the route of administration, and the patient / subject's condition. For example, the appropriate dose of the composition of the present invention for a given patient / subject is determined by the attending physician (or the person administering the composition) considering various factors known to alter the action of the composition of the present invention, such as the severity and type of hematological malignancy, body weight, sex, diet, timing and route of administration, other medications, and other relevant clinical factors. Dosage and administration schedule may be modified depending on the patient / subject's overall condition, specific condition, disorder, or symptoms.

[0133] The pharmaceutical compositions described herein are, advantageously, provided in unit dosage forms.

[0134] In one example, human albumin (e.g., serum albumin or recombinant human albumin) is intended to be administered at multiple intervals as part of a multi-dose regimen. A suitable multi-dose regimen may be identified by those skilled in the art. For example, a multi-dose regimen may include multiple divided doses administered at equal intervals of approximately 1 to 30 days. In one example, a multi-dose regimen may include multiple divided doses administered at unequal intervals of approximately 1 to 30 days. In another example, a multi-dose regimen may include two or more doses until a total cumulative dose is reached. In some cases, human albumin may be administered at equal or unequal intervals of approximately 1 to 6 days.

[0135] The compositions described herein contain appropriate levels (amount, concentration, or dose) of human albumin. In the context of any of the dosing regimens described herein, the human albumin in the composition may be intended to be administered in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval when it is part of a multi-dose regimen (for example, when it is part of any of the multi-dose regimens specifically described herein). For example, the human albumin in this composition may be intended to be administered in individual doses ranging from approximately 20 g / interval to approximately 200 g / interval when it is part of a multi-dose regimen (for example, when it is part of any of the multi-dose regimens specifically described herein). For example, 40 g / interval. For example, 40 g / day.

[0136] In certain cases, a multi-dose regimen may include multiple divided doses of the composition described herein, administered at equal intervals of approximately 1 to approximately 30 days, with the human albumin in the composition being administered in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval (e.g., approximately 20 g / interval to approximately 200 g / interval). In other certain cases, a multi-dose regimen may include multiple divided doses of the composition described herein, administered at unequal intervals of approximately 1 to approximately 30 days, with the human albumin in the composition being administered in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval (e.g., approximately 20 g / interval to approximately 200 g / interval). In yet another certain case, a multi-dose regimen may include two or more doses of the composition described herein until a total cumulative dose is reached, with the human albumin in the composition being administered in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval (e.g., approximately 20 g / interval to approximately 200 g / interval). In some cases, a multi-dose regimen may involve administering human albumin at approximately 40 g / day intervals. In some cases, a multi-dose regimen may involve administering human albumin at approximately 40 g / day intervals. In some cases, a dosing regimen may involve administering human albumin at approximately 40 g / day intervals over 2 to 6 days. For example, a dosing regimen may involve administering human albumin at approximately 40 g / day over 2 to 6 days. In some cases, a dosing regimen may involve administering human albumin at approximately 40 g / day over a median of 2.5 days.

[0137] In each of these examples, the dosing interval may be every 15 days or less. For example, the dosing interval may be every 10 days or less.

[0138] In certain examples, the compositions described herein contain human albumin at a concentration between approximately 4% and 25% (w / v) [e.g., approximately 20% (w / v)]. In other words, the doses and administration regimens provided above can be achieved using compositions containing human albumin, with a concentration of human albumin between approximately 4% and 25% (w / v) [e.g., approximately 20% (w / v)].

[0139] The compositions described herein are particularly useful for the treatment of subjects. The terms “treatment” or “to treat” mean any treatment of a disease or disorder in a subject such as a mammal, and include prevention or defense of the disease or disorder, i.e., preventing the manifestation of clinical symptoms; inhibition of the disease or disorder, i.e., stopping or suppressing the manifestation of clinical symptoms; and / or alleviation of the disease or disorder, i.e., causing regression of clinical symptoms. In some examples, the term treatment is used to treat deficient immune cell function. In some embodiments, the term “treatment” is used to treat a disease selected from the group consisting of systemic inflammatory response syndrome, decompensated cirrhosis, acute hepatic failure, the pre-stage of acute exacerbation of chronic hepatic failure, acute exacerbation of chronic hepatic 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.

[0140] Those skilled in the art will understand that in human medicine, the ultimate triggering event may be unknown, latent, or the patient may only be identified long after the event has occurred, and therefore, it is not always possible to distinguish between "prevention" and "suppression." Accordingly, as used herein, the term "prevention of onset" is intended to encompass both "prevention" and "suppression" as defined herein, as components of "treatment."

[0141] As used herein, systemic inflammatory response syndrome (SIRS) refers to an inflammatory condition resulting from infectious or non-infectious causes. SIRS is a serious condition associated with systemic inflammation, organ damage, and organ failure. It is a subset of cytokine storms, involving abnormal regulation of various cytokines. SIRS is also closely associated with sepsis in patients who meet the criteria for SIRS and have a suspected or confirmed infection. Patients with SIRS may be identified using the following criteria:

[0142] In adults, symptoms of SIRS include, but are not limited to, the following: (i) Body temperature is below 36°C (96.8°F) or above 38°C (100.4°F) (ii) Heart rate exceeds 90 beats per minute (iii) Tachypnea (increased respiratory rate) exceeding 20 breaths per minute; or arterial carbon dioxide partial pressure less than 4.3 kPa (32 mmHg) (iv) White blood cell count of 4000 cells / mm 3 (4×10 9 Less than 12,000 cells / mm³ (individual cells / L) 3 (12×10 9 If the percentage exceeds 10% of individual cells / L, or if more than 10% are immature neutrophils (band type), it is called bandemia or "left shift".

[0143] Regardless of whether or not there are signs of infection, a patient may be diagnosed with "SIRS" if they meet two or more of these criteria. Patients with SIRS and acute organ dysfunction may be referred to as having "severe SIRS."

[0144] In children, the SIRS criteria are modified as follows: (i) A heart rate that is two standard deviations above the age-specific reference range, or a sustained high heart rate of unknown cause lasting more than 30 minutes but up to 4 hours, in the absence of stimuli such as pain or drug administration. In infants, this also includes a heart rate below the age-specific 10th percentile, or a sustained low heart rate of unknown cause lasting more than 30 minutes, in the absence of vagal nerve stimulation, beta-blockers, or congenital heart disease. (ii) Body temperature measured from the oral cavity, rectum, Foley catheter probe, or central venous catheter probe is below 36°C or above 38.5°C. (iii) Respiratory rate more than 2 standard deviations higher than the age-specific reference value, or need for mechanical ventilation unrelated to neuromuscular disease or anesthesia administration. (iv) An increase or decrease in white blood cell count relative to age that is not related to chemotherapy, or a band greater than 10% or other immature morphology. In pediatric patients, SIRS requires an abnormal body temperature or white blood cell count.

[0145] SIRS may be associated with subjects having diseases selected from the group consisting of decompensated cirrhosis, acute liver failure, pre-ACLF (pre-acute exacerbation of chronic liver failure), acute exacerbation of chronic liver failure (ACLF), 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. Therefore, SIRS may be present in subjects having diseases selected from the group consisting of decompensated cirrhosis, acute liver failure, pre-ACLF (pre-acute exacerbation of chronic liver failure), acute exacerbation of chronic liver failure (ACLF), 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. Accordingly, compositions containing human albumin as described herein may be used to treat SIRS in diseases selected from the group consisting of decompensated cirrhosis, acute hepatic failure, pre-ACLF, acute exacerbation of chronic hepatic failure (ACLF), 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, these compositions may be used to treat aspects of SIRS in the described diseases). Methods for identifying whether a subject has one or more of these diseases are well known.

[0146] In one example, the subjects are patients with sepsis or hepatic conditions. Therefore, subjects with SIRS may have SIRS related to sepsis or hepatic conditions. Examples of patients with sepsis include patients with severe sepsis or septic shock. Examples of patients with hepatic conditions include patients with decompensated cirrhosis, acute hepatic failure, the pre-acute exacerbation of chronic hepatic failure (pre-ACLF), and / or acute exacerbation of chronic hepatic failure (ACLF).

[0147] In one example, the subject has decompensated cirrhosis. Decompensated cirrhosis is defined as an acute deterioration of liver function in a patient with cirrhosis, characterized by the recent onset of ascites, hepatic encephalopathy, gastrointestinal bleeding, or any combination thereof. In this specification, decompensated cirrhosis is also referred to as "acute decompensated cirrhosis," "acutely decompensated cirrhosis," or "decompensated liver disease," and these terms may be used interchangeably.

[0148] Among patients with acute decompensated cirrhosis, immune activation is more pronounced in those with acute exacerbation of chronic liver failure (ACLF), a syndrome characterized by severe systemic inflammation, single multi-organ dysfunction / failure, and a high incidence 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), and this tendency is more pronounced in patients with ACLF compared to those without ACLF (Weiss et al., 2021, Frontiers in Immunol, vol. 12, art. 699563). Furthermore, the PREDICT study recently identified a subset of subjects presenting with severe systemic inflammation but without developing ACLF, who were at high risk of developing ACLF and at high risk of death during hospitalization or within weeks after discharge (Trebicka et al., 2020, J Hepatol, 73:842-854). This subset of subjects is classified as having pre-ACLF acute decompensated cirrhosis.

[0149] Pre-ACLF is defined as a group of patients with acute decompensated cirrhosis who were admitted to the hospital without ACLF but developed ACLF during hospitalization or within 3 months after admission. Pre-ACLF can be classified into two distinct phenotypes: “early” pre-ACLF, which defines patients who develop ACLF during their initial hospitalization, and “late” pre-ACLF, which defines patients who develop ACLF between discharge from initial hospitalization and the end of the 3-month follow-up period. Patients admitted with “early” pre-ACLF show severe systemic inflammation, a feature that is moderate in “slowly progressive” pre-ACLF patients.

[0150] In this specification, subjects with acute decompensated cirrhosis accompanied by ACLF are also referred to as ACLF subjects or ACLF patients. In this specification, subjects with pre-ACLF acute decompensated cirrhosis are also referred to as pre-ACLF subjects or pre-ACLF patients.

[0151] In one example, the subjects are ACLF subjects or pre-ACLF subjects. In other words, the subjects may have ACLF decompensated cirrhosis (also referred to herein as ACLF acute decompensated cirrhosis) or pre-ACLF decompensated cirrhosis (also referred to herein as pre-ACLF acute decompensated cirrhosis). In one example, pre-ACLF decompensated cirrhosis is early pre-ACLF decompensated cirrhosis.

[0152] In one case, the subject has liver failure. Acute liver failure is characterized by acute liver injury, hepatic encephalopathy (changes in mental state), and an elevated prothrombin time / international normalized ratio (INR). It is also called fulminant liver failure, acute hepatic necrosis, fulminant hepatic necrosis, or fulminant hepatitis.

[0153] Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled 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 those skilled in the art with a general dictionary of many of the terms used in this invention. Any methods and materials similar or equivalent to those described herein may be used in the practice of this invention, but preferred methods and materials are described herein. Thus, the terms defined immediately below are explained in more detail by referring to the entire specification. Also, where used herein, the singular terms “a,” “an,” and “the” include the plural form unless the context clearly indicates otherwise. Unless otherwise specified, nucleic acids are written from left to right in the 5' to 3' direction, and amino acid sequences are written from left to right in the amino group to carboxyl group direction. It should be understood that the present invention is not limited to the specific methodologies, protocols, and reagents described, which may vary depending on the context in which they are used by those skilled in the art.

[0154] Aspects of the present invention are illustrated by the following non-limiting embodiments. [Examples]

[0155] [Example 1] The effect of albumin therapy on circulating immune cell gene signatures in patients with acute decompensated cirrhosis (effect of albumin on circulating immune cells). background Evaluating genome-wide RNA expression in the blood is a valuable tool for simultaneously investigating the signatures of different types of circulating immune cells. 5、19~21The inventors report the results of a discovery study in which whole blood RNA-seq data was obtained twice from 49 patients in the PREDICT study. At the time of the first acquisition, these patients were in the pre-ACLF stage (at admission), and at the time of the second acquisition, these patients had progressed to the ACLF stage within a few days of admission during their first hospitalization. Therefore, all patients were in the "early" form of pre-ACLF. Since 30 patients received albumin during ACLF progression, while the remaining 19 did not, the discovery study was the first approach to capture the in vivo effect of albumin on blood immune cell-related gene signatures. Both the longitudinal design and the well-established similarity between the intensity of systemic inflammation in "early" pre-ACLF and ACLF should minimize inter-individual variability in gene expression, which makes it difficult to capture the in vivo effect of albumin on blood immune cells.

[0156] Surprisingly, the discovery study revealed a specific increase in gene signatures, primarily associated with B cells, plasma cells, and neutrophils, in patients treated with albumin. Based on these findings, the inventors conducted additional studies, including further investigation of the ex vivo effects of albumin on newly isolated peripheral blood mononuclear cells (PBMCs) and newly isolated serum neutrophils in new patients with acute decompensated cirrhosis.

[0157] method Patient and study design All patients (i.e., 64 patients) had acute decompensated cirrhosis without ACLF at the time of enrollment. Each patient provided written informed consent to participate. Each study described below was pre-approved by the Ethics Committee of the Hospital Clinic, Barcelona, ​​Spain.

[0158] Discovery Research The inventors first carefully selected PREDICT studies based on the following criteria. 3The following 49 patients were examined: i) All patients were “early” pre-ACLF at enrollment (time point 1 [T1]) because they developed ACLF during their initial hospitalization (time point 2 [T2]; data not shown). For simplicity, the inventors will use the term “pre-ACLF” instead of “early pre-ACLF” below. The inventors hypothesized that by examining patients exhibiting the most severe forms of acute decompensated cirrhosis using longitudinal data, they could limit the impact of interpersonal variability in the assessment of albumin effect. ii) In all patients, whole blood RNA-seq data were obtained at T1 and at T2 as prespecified in the protocol. iii) None of the 49 patients received albumin before T1. iv) Thirty of the 49 patients received albumin between T1 and T2, forming the albumin group, while the remaining 19 did not receive albumin between T1 and T2, forming the non-albumin group (data not shown). Of particular note is that the inventors also obtained clinical blood cell count results and whole blood RNA-seq data from 10 age-matched healthy subjects (HS) (data not shown). The inventors analyzed longitudinal whole blood RNA-seq data obtained from 49 patients for two purposes. The first purpose was to evaluate whether the transcriptome characteristics of the 49 patients investigated at the pre-ACLF stage differed from those of the same patients at the time of progression to ACLF, regardless of whether albumin was administered. In fact, unlike the whole blood transcriptome characteristics of ACLF reported to date, 5 The characteristics of pre-ACLF have not yet been investigated. The second primary objective was to evaluate the potential influence of albumin on the RNA signature of circulating immune cells through blood transcriptome analysis of albumin and non-albumin groups.

[0159] Additional research Following the results of the discovery study, the inventors enrolled nine new patients with acute decompensated cirrhosis and isolated peripheral blood mononuclear cells (PBMCs) from these patients. They then investigated the ex vivo effect of albumin on the cells constituting the PBMCs using single-cell RNA-seq (scRNA-seq). Furthermore, they isolated neutrophils from an additional set of six new patients and studied the ex vivo effect of albumin on the major antimicrobial function of neutrophils.

[0160] RNA preparation for whole blood RNA-seq As mentioned above 5、19 RNA was isolated from blood stored in Tempus tubes using the Tempus® Spin RNA Isolation Kit (product number 43802, Applied Biosystems, Foster City, CA). Importantly, using the HUGO Genome Nomenclature Committee, an approved resource for human gene nomenclature, the inventors designed a comprehensive protocol that included not only protein-coding genes but also genes in 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.

[0161] Whole blood RNA-Seq analysis Evaluation of differential gene expression. Differentially expressed genes (DEGs) were defined as those that simultaneously meet two criteria: an absolute ploidy (FC) change greater than 1.5 times and an adjusted p-value less than 0.05.

[0162] Evaluation of immune cell signatures using SingleR R software SingleR Software 22 Using this method, whole blood RNA-seq data from research participants were compared to a reference dataset (GSE107011) containing 114 human bulk RNA-seq samples from immune cell populations fractionated from four healthy subjects. 23This reference dataset contained 114 human RNA-seq samples annotated with 10 major immune cell types. The dataset (GSE107011) also included samples annotated with 29 further subdivided immune cell types. The SingleR pipeline compared the whole blood RNA-seq data first with the 10 major immune cell types used as a reference, and then with the 29 subdivided immune cell types.

[0163] Gene set enrichment analysis QuSAGE package 24 Using the QuSAGE quantitative gene expression analysis method implemented in [platform name], gene set enrichment analysis was performed using 346 blood transcription modules (BTMs) as the gene set. BTMs are sets of gene modules developed through the integration of a large network of publicly available human blood transcriptomes. 20 QuSAGE provides activity scores for each gene set in each comparison (e.g., pre-ACLF vs. healthy subjects, T2 vs. T1 in the albumin group). BTM was identified as differentially expressed if its QuSAGE activity score was less than 0.05, or, where appropriate, less than 0.05.

[0164] Analysis of shared genes and shared BTMs DEGs or differentially expressed BTMs were defined as being shared between the two comparisons if the signs of the DEGs or differentially expressed BTMs matched.

[0165] ScRNA-seq in PBMCs Peripheral venous blood (8 ml) was collected by venous puncture from 9 patients and 4 age-matched healthy subjects into sterile, pyrogen-free tubes containing EDTA (Becton Dickinson, East Rutherford, NJ). (Previously described method) 25 PBMCs were immediately isolated according to the procedure. PBMCs were divided into 1.5 × 10⁻⁶ units. 6Cells were seeded at a density of individual cells / ml and incubated with albumin (Albutein®, Grifols, Barcelona, ​​Spain) (15 mg / ml) or a medium in a 37°C, 5% CO2 incubator for 2 hours. After culturing, cells were rapidly transferred (within 30 minutes) to ice using the CNAG Single Cell Genomics platform.

[0166] Experiments in neutrophils Neutrophils were isolated from peripheral venous blood (20 ml) collected from six patients and five age-matched healthy subjects using the Ficoll-Hypaque method. The isolated neutrophils were immediately used in various experiments to evaluate their major antimicrobial functions, such as degranulation assays and phagocytic assays.

[0167] result Characterization of pre-ACLF at T1 and ACLF at T2 in 49 patients in the discovery cohort. Clinical characteristics The inventors first characterized all patients at time T1 (pre-ACLF) and time T2 (when ACLF progressed). The characteristics of both groups were similar in standard laboratory values ​​(including clinically differentiated blood cell counts of neutrophils, monocytes, and lymphocytes; data not shown) and, although data not shown, in blood levels of inflammatory markers. Notably, in both pre-ACLF and ACLF, blood levels of inflammatory markers were elevated compared to corresponding levels in healthy subjects (data not shown). The only two differences were that, as expected, organ failure was observed in ACLF at T2 but not in pre-ACLF at T1, and that white blood cell counts were higher in ACLF than in pre-ACLF (data not shown). Compared to healthy subjects, pre-ACLF and expected ACLF 5The cells were characterized by leukocytes, neutrophils, and lymphocytes (data not shown). Notably, the inventors found a strong positive correlation between clinical blood cell counts and blood cell counts estimated from RNA-seq (data not shown), suggesting that RNA-seq analysis of whole blood RNA can provide information that correlates with baseline clinical measurements of blood cell counts.

[0168] Next, the inventors examined the characteristics of T1 and T2 time points in patients in the albumin group and patients in the non-albumin group. In the albumin group, the median time from T1 to T2 for blood sampling for RNA-seq was 11.5 days (interquartile range [IQR], 6.2–18.7).

[0169] The median duration of albumin treatment was 2.5 days (IQR, 2-6), and the albumin dose was 40.0 g per day (IQR, 22.5-57.5). The interval between the final albumin administration and subsequent blood collection for RNA-seq was 1.5 days (IQR, 0.0-2.7), indicating that the inventors investigated patients immediately after albumin administration. Albumin was administered based on standard medical indications. In the non-albumin group, the median time from T1 to T2 was 29 days (IQR, 16-52).

[0170] In both the albumin and non-albumin groups, progression from pre-ACLF (T1) to ACLF (T2) was associated with a significant increase in the MELD score. However, with the exception of elevated serum creatinine at T2, no significant changes were observed within the groups in other longitudinally collected standard laboratory data and circulating levels of 14 inflammatory mediators (Table 1). Some inter-group differences were observed, however. The delay between T1 and T2 was shorter in the albumin group than in the non-albumin group. Serum levels of C-reactive protein and interleukin-6 were higher in the albumin group at both T1 and T2. Patients in the albumin group had a higher mortality rate by days 28 and 90 than patients in the non-albumin group, but this difference was predictable considering the higher frequency of SBP and HRS-AKI in the albumin group.

[0171] Transcriptional characteristics of all patients in the pre-ACLF stage (T1) and ACLF stage (T2) Analysis of DEGs in two comparisons—pre-ACLF versus healthy subjects and ACLF versus healthy subjects—resulted in a strong similarity between the two disease stages, as expected. Consequently, the number of DEGs was high in both comparisons, specifically 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 DEGs assigned to ACLF (data not shown). Furthermore, when the inventors compared the change in effect size (log2 multiplier change [FC]) between pre-ACLF versus healthy subjects and ACLF versus healthy subjects, they found a high degree of agreement in the magnitude of change between the two signatures (data not shown).

[0172] Next, the inventors analyzed differential expression of blood gene modules (BTMs) using QuSAGE. 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, 20 The 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). This result suggests widespread changes in the blood transcription module space in both pre-ACLF and ACLF. Notably, in both pre-ACLF (12 modules) and ACLF (8 modules), only 20 modules showed specific differential expression compared to healthy subjects, while 179 DE BTMs were shared, meaning there are DE modules with the same sign 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 identified (data not shown). Shared upregulated BTMs were associated with innate immunity, including those related to TLRs and inflammatory signaling, interferon-alpha response, and innate immune cells such as neutrophils, monocytes, and dendritic cells. Shared downregulatory BTMs were associated with T cells, NK cells, and antigen presentation. These results were consistent with those obtained when analyzing the gene signatures of 10 major immune cell types and 29 subdivided immune cell types using SingleR R software. Overall, analysis of whole blood RNA-seq data using QuSAGE and SingleR R software provided consistent results highlighting the similarities in the blood transcription module landscape between pre-ACLF and ACLF. Both disease stages were characterized by the simultaneity between increased innate immune cell-related gene signatures and decreased adaptive immune cell-related gene signatures.

[0173] Whole blood gene signatures associated with albumin therapy The inventors analyzed differential gene expression between T2 and T1 in the albumin group and the non-albumin group and found that the gene signatures differed between the two groups. In fact, there were 269 DEGs specific to the albumin group (92 upregulatory genes and 177 downregulatory genes), while there were 103 DEGs specific to the non-albumin group (64 upregulatory genes and 39 downregulatory genes), with only 36 DEGs having matching signs common to both groups (Figure 1A). The identification of DEGs in the volcano plot (Figure 1B) showed group specificity for both upregulatory and downregulatory genes. For example, a variety of immunoglobulin genes (see below) and major neutrophil-related genes (CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, MCEMP1) were upregulatory genes in the albumin group in the T2 vs. T1 transition (Figure 1B upper), whereas these genes were not present in the upregulatory gene group in the non-albumin group (Figure 1B lower).

[0174] Since immunoglobulin genes were identified through DEG analysis (Figure 1B), the inventors compared the effect size changes (log2 multiplier changes) between T2 and T1 within each group for 125 genes (including 120 immunoglobulin genes) in the Gene Ontology gene set titled "GOCC Immunoglobulin Complex." As a result, the inventors found that the effect size change was larger in the albumin group compared to the non-albumin group, and the number of upregulated immunoglobulin genes was 33 and 6, respectively (Figure 1C). In the albumin group, genes encoding the constant region of the immunoglobulin heavy chain (IGHM, IHG2, IHG3, IHG4, IGHA2) were specifically upregulated. These findings are consistent with the results of SingleR analysis, which showed an increase in the gene signature of plasmablasts specific to the albumin group (Figure 1D, top). In the albumin group, immunoglobulin genes specifically upregulated included the constant region genes of immunoglobulin light chain kappa (IGKC) and lambda (IGLC1), the V region gene of the immunoglobulin heavy chain variable domain, and the V region gene of the variable domains of light chain kappa and lambda. In the non-albumin group, three genes specifically upregulated were genes encoding the V region of the immunoglobulin heavy chain variable domain (Figure 1C). Taken together, these findings suggest broad upregulation of immunoglobulin-encoding genes specific to patients who received albumin.

[0175] Next, the inventors applied QuSAGE to identify BTMs showing differential expression between T2 and T1 within each group. First, the inventors observed that the number of differentially expressed BTMs was greater in the albumin group than in the non-albumin group (36 vs. 6, respectively). Only one differentially expressed BTM was specific to the non-albumin group, and this BTM was upregulated and related to the endoplasmic reticulum (Figure 1E). In contrast, 31 differentially expressed BTMs (13 with decreased expression, 18 with increased expression) were specific to the albumin group (Figure 1E). The downregulated modules were related to either erythrogenesis, the cytoskeleton, or intercellular junctions (Figure 1E). Among the albumin-specific upregulated BTMs were modules related to innate immunity and inflammation, including modules related to activated dendritic cells, complement and other receptors in DCs, pro-inflammatory cytokines, and chemokines (Figure 1E). Albumin-specific upregulated BTMs included B cell-related modules (including B cell, plasma cell, and immunoglobulin-rich modules), as well as mismatch repair, cell cycle, and mitosis-related modules. Finally, BTMs related to transcriptional networks of transcription factors (TFs), such as c-Myc and members of the E2F family, were specifically upregulated in the albumin group (Figure 1E). This finding suggests increased activity of these TFs in the blood of patients treated with albumin. Since c-Myc and members of the E2F family are cell cycle regulators, their increased activity may explain the induction of cell cycle and mitosis-related gene modules observed in the blood of patients treated with albumin.

[0176] Notably, the RNA-seq predictive signatures of T cells and NK cells (Figure 1D), as well as the gene modules associated with these cells, remained downregulated in all albumin-treated patients, although downregulation is characteristic of ACLF (and pre-ACLF; data not shown). Nevertheless, we observed that the BTM titled "Mitogenic Cell Cycle (M4.11) in Stimulated CD4 T Cells" was specifically upregulated in albumin-treated patients (Figure 1E), suggesting that transcription is activated in some way in the CD4 T cells of these patients. Consistent with this, we found no overlap between the constituent genes of the BTM (M4.11) related to the mitogenic cell cycle in stimulated CD4 T cells and the genes used in the signature analysis of CD4 T cells using SingleR software. In summary, the results revealed that the blood of patients who received albumin administration was characterized by increased signatures related to immune cells such as B cells, plasma cells, some innate immune cells, and possibly CD4 T cells, which were associated with increased signatures for mitosis and the cell cycle, as well as increased signatures indicating elevated activity of E2F family transcription factors.

[0177] ScRNA-seq of human PBMCs to investigate the ex vivo effects of albumin Based on the results of whole blood RNA-seq analysis, the inventors investigated the ex vivo effect of albumin on human PBMCs using scRNA-seq. Fresh PBMCs collected from 9 patients with acute decompensated cirrhosis and 4 healthy controls were ex vivo exposed to albumin (15 mg / ml) and a medium for 2 hours before performing the scRNA-seq experiment.

[0178] The inventors analyzed a total of 66,064 human PBMCs derived from patients and healthy subjects and clustered them into three major immune cell lineages, including B lymphocytes, myeloid cells, and T cells. Furthermore, the inventors subclustered each lineage using standard genetic markers and annotated them with different cell types.

[0179] The inventors analyzed 3,786 B lymphocytes (1,946 from patients and 1,840 from healthy subjects) and identified 10 major populations. The results for patient cells are shown in Figure 2A. Notably, we describe two subpopulations that acquired a transitional-like B cell profile characterized by higher expression of CD79B, IGHD, and TCL1A (transitional 1B cells) and CD55 (transitional 2B cells). The inventors found that in patient cells exposed to albumin, the abundance of cells expressing the transitional-like B cell signature increased significantly and naive B cells decreased (Figures 2B and 2C), while other B lymphocyte populations remained unchanged. Furthermore, the inventors further evaluated 26,898 myeloid cells (21,677 patient-derived cells and 5,053 healthy control-derived cells, respectively) and found that they were clustered into 13 different cell populations: 7 clusters were monocyte-related, 3 clusters were dendritic cell (DC)-related, and the other 3 clusters included myeloid suppressor cells and granulocyte-monocyte progenitor cells. The results for patient cells are shown in Figure 2D. The inventors found that intermediate monocyte HAVCR2 was present in patient cells exposed to albumin. + We observed a significant increase in the abundance of plasmacytoid dendritic cells (DCs) (Figures 2E and 2F), while the abundance of other myeloid cell populations remained unchanged.

[0180] Based on differential expression-based BTMs found in the T cell compartment, the inventors also analyzed 35,380 T lymphocytes, including 21,731 CD4 T cells (12,692 patient-derived and 9,039 healthy control-derived, respectively), 10,072 CD8 T cells (3,483 patient-derived and 6,589 healthy control-derived, respectively), and 3,577 non-conventional T cells (1,292 patient-derived and 2,285 healthy control-derived, respectively). CD4 T cells were clustered into 10 different cell populations. The results for patient cells are shown in Figure 2G. The inventors observed that albumin exposure in patients altered the profile of the CD4 T cell compartment (Figure 2H), significantly decreasing the abundance of activated memory CD4 T cells and increasing central memory ITGB1+ CD4 T cells (Figure 2I). CD8 T cells and non-conventional T cells were clustered into 13 different cell populations, but no significant changes were observed in the cell populations after albumin exposure.

[0181] One BTM that was specifically upregulated in patients who received albumin was the BTM titled "Mitotic Cell Cycle in Stimulated CD4 T Cells (M4.11)" (Figure 1E). To validate the M4.11 signature in scRNA-seq, we calculated the gene signature score of this BTM across all CD4 T cells using the Ucell package. 26 The M4.11BTM signature exhibited different distribution patterns on CD4 T cells under albumin-treated and medium conditions, showing significantly higher cell density on albumin-treated cells compared to medium cells (Figure 2J).

[0182] Furthermore, analysis of PBMCs derived from healthy subjects showed similar albumin effects in both the B lymphocyte and myeloid cell compartments as those from patients.

[0183] Investigation of the ex vivo effects of albumin on neutrophils Based on the analysis of whole blood RNA-seq data from albumin and non-albumin groups, the inventors focused on neutrophils. In fact, pairwise comparison of whole blood RNA-seq data between T2 and T1 neutrophils in the albumin and non-albumin groups revealed upregulation of nine major neutrophil genes specific to patients receiving albumin (Figure 1B). Therefore, based on these findings, the inventors designed an ex vivo study using newly isolated neutrophils. Because neutrophils provide the first line of defense against most pathogens and express fewer genes than any other leukocyte, this study focused on analyzing the effects of albumin on neutrophil function.

[0184] During the host defense response, neutrophils release myeloperoxidase (MPO), an essential antimicrobial protein that is mainly localized in azurofil granules or primary neutrophil granules. 27 Therefore, neutrophil degranulation can be quantified by measuring MPO activity. Figure 3A shows enhanced MPO activity in the supernatant when neutrophils from patients with acute decompensated cirrhosis and healthy subjects were incubated with serum albumin or recombinant human albumin.

[0185] Phagocytosis is another defense mechanism against pathogens and can be assessed by measuring the uptake of fluorescently labeled zymosan particles by neutrophils. Figure 3B shows increased zymosan phagocytosis by neutrophils from both patients with acute decompensated cirrhosis and healthy subjects incubated with serum albumin or recombinant human albumin.

[0186] Consideration This discovery study was the first to explore the mechanism of action of albumin in patients with acute decompensated cirrhosis using longitudinal genomics. The design of this study, particularly the selection of candidates and aspects related to albumin treatment, was formulated considering both known and unpublished findings. The degree of systemic inflammation changes significantly. 2Consequently, ACLF hospitalized patients, who are known to exhibit extremely dynamic clinical courses such as improvement, stabilization, or deterioration within a few days of hospitalization, are clearly not eligible subjects for this study, which aims 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 excluded because they involve moderate systemic inflammation, which may further improve during hospitalization. To better evaluate the potential of albumin-induced changes in immune cell gene expression, the inventors intended to include only patients with severe systemic inflammation. For this reason, the inventors focused on patients with "early" pre-ACLF, where the degree of systemic inflammation is similar to that observed in ACLF. Despite the bias that patients who receive albumin therapy tend to have a higher severity of disease, the inventors decided to focus only on patients who received albumin therapy for well-established indications (puncture treatment, prevention of SBP-related HRS-AKI, or treatment of HRS-AKI). 13 The inventors excluded patients who had received albumin within one month prior to T1, patients whose two whole blood RNA-seq evaluations were too far apart, and patients whose interval from the last albumin dose to T2 was more than 10 days. Finally, although albumin dosage and treatment duration varied by indication, the inventors decided to analyze data from all patients regardless of albumin dosage. Of the 1273 patients enrolled in the PREDICT study, 49 patients met all the selection criteria and were therefore included in this discovery study.

[0187] The inventors first compared the blood transcriptomes of all patients at each time point with those obtained in healthy subjects. They observed that pre-ACLF and ACLF were characterized by increased gene signatures associated with innate immune cells (including neutrophils, monocytes, and dendritic cells) and decreased gene signatures associated with lymphocytes (including T cells, B cells, and NK cells). These findings suggest that in the blood of patients with cirrhosis and severe systemic inflammation, simultaneous activation of innate immunity via myeloid cells and depletion of lymphocyte compartments occur. The similarities between pre-ACLF and ACLF in most clinical characteristics, standard laboratory values, and blood cytokine levels, as well as transcriptional properties, supported the inventors' hypothesis that the subjects were in a relatively stable state of systemic inflammation during the study period. This stable state was a prerequisite for avoiding confounding factors that would mask the in vivo effects of albumin on blood immune cells.

[0188] The key findings of the discovery study were provided by a comparison of T2 vs. T1 blood transcriptomes in 30 patients who received albumin and 19 patients who did not. The inventors observed that patients treated with albumin exhibited genetic signatures not observed in patients who did not receive this treatment. Thus, patients treated with albumin were characterized by a specific increase in signatures related to adaptive immune cells (including B cells, plasma cells, immunoglobulins, and CD4 T cell-related signatures) and innate immune mononuclear myeloid cells. Furthermore, TF-related signatures such as cell cycle, mitosis, c-Myc, and E2F family members were also specifically increased in patients treated with albumin. Importantly, scRNA-seq results in PBMCs from patients ex vivo exposed to albumin indicated that albumin itself induces alterations in adaptive and innate immune cells. These findings suggest that albumin administration induces signals that lead to the expansion of the B cell compartment and the CD4 T cell compartment, while simultaneously activating certain mononuclear myeloid cells.

[0189] Another major finding in this study was the activation of neutrophils by albumin. In patients treated with albumin, several genes that marker activated low-density neutrophils were upregulated. Furthermore, ex vivo experiments significantly demonstrated that neutrophils from patients exposed to albumin showed increased degranulation response and enhanced phagocytosis. Therefore, albumin restores neutrophil-mediated defense mechanisms, which are known to be significantly impaired in acute decompensated cirrhosis. 5、28 .

[0190] The prevalence of bacterial infections at hospitalization in patients with acute decompensated cirrhosis is very high, at 37.3% in patients with ACLF. 11 In patients without ACLF, the figure was 25.1%. 3、4 Even among patients who were infection-free upon admission, 46% of patients presented with ACLF. 11 , and 18% of patients who did not present with ACLF 4The patient developed a bacterial infection during hospitalization. These data indicate a significant impairment of the defense mechanisms against microorganisms. Severe damage to innate immune cells (indicated by the deficient antimicrobial function of neutrophils and monocytes) and depletion of the lymphocyte compartment (including T cells, B cells, and NK cells), along with the presence of enteric bacterial migration, likely explain the increased risk of infection in patients with acute decompensated cirrhosis. 29 The results of this study strongly suggest that albumin can restore both the deficient antimicrobial function of neutrophils and the depleted lymphocyte compartment in the most severe forms of acute decompensated cirrhosis.

[0191] An interesting observation from the ex vivo studies presented herein is that albumin responses in lymphocytes (scRNA-seq experiments) and neutrophil responses (experiments evaluating degranulation and phagocytosis) occurred in less than two hours, indicating that the signal mediated by albumin molecules was transmitted extremely rapidly within the target cells.

[0192] In conclusion, in patients with severe acute decompensated cirrhosis, the inventors advantageously demonstrated that albumin promotes the expansion of the B cell compartment and the CD4 T cell compartment, acts on mononuclear myeloid cells, and resets the antimicrobial function of neutrophils to normal. Therefore, advantageously, when human albumin is administered to patients with decompensated cirrhosis, clinical benefits such as prevention of infection and suppression of PAMP-induced systemic inflammation may be obtained in patients with decompensated cirrhosis.

[0193] Abbreviation ACLF, acute exacerbation of chronic liver failure; AKI, acute kidney injury; FDR, false detection rate; HRS, hepatorenal syndrome; NK, natural killer; PAMP, pathogen-associated molecular pattern; PBMC, peripheral blood mononuclear cells; QuSAGE, quantitative set analysis of gene expression; RNA-seq, RNA sequencing; scRNA-seq, single-cell RNA-seq; UMAP, homogeneous manifold approximation and projection.

[0194] [Table 1-1] [Table 2-2]

[0195] Additional materials and methods Neutrophils were isolated from peripheral venous blood (20 ml) collected from six patients with acute decompensated cirrhosis (AD) and five age-matched healthy subjects using the Ficoll-Hypaque method. Briefly, patients with acute decompensated cirrhosis were registered in the liver intensive care unit of Hospital Clinic (Barcelona, ​​Spain). Blood from four healthy donors was obtained in agreement with the Hospital Clinic blood bank. Peripheral venous blood (8 ml) was collected by venipuncture and collected in sterile pyrogen-free EDTA-added tubing (Becton Dickinson, Grenoble, France). Blood samples were centrifuged at 200 g for 10 minutes to collect plasma. Precipitated cells were diluted to a volume of 20 ml with DPBS- / -. The diluted blood was overlaid on 13.3 ml of Ficoll-Hypaque and centrifuged at 500 g for 25 minutes with break-off. After separating the PBMC layer from the supernatant, neutrophils were collected from the precipitate and incubated in ammonium chloride-potassium lysis buffer preheated to room temperature for 10 minutes to remove erythrocytes. The precipitate was then centrifuged at 400 g for 5 minutes. The erythrocyte lysis procedure was repeated twice, and the resulting precipitate was washed with DPBS. The isolated neutrophils were resuspended in RPMI1640 medium [fetal bovine serum (FBS)-free] containing penicillin (100 U / mL), streptomycin (100 U / mL), and L-glutamine (4 mM), and used in subsequent neutrophil function assays.

[0196] Degranulation assay. After standing for 30 minutes, neutrophils were divided into 3 × 10⁻¹⁴ cells. 6Cells were seeded at a density of individual cells / mL and incubated for 2 hours at 37°C in a 5% CO2 incubator with or without phorbol-12-myristate-13-acetate (100 nM), along with either albumin, recombinant human albumin (both 15 mg / ml), or a media control. After incubation, the supernatant was collected and degranulation was measured using the Neutrophil Myeloperoxidase Activity Assay Kit (Cayman Chemical, AnnArbor, MI). Briefly, 25 μl of neutrophil supernatant and 25 μl of assay buffer were added together to each well of an experimental plate. Next, 50 μl of 3,3',5,5'-tetramethylbenzidine (TMB), a substrate for horseradish peroxidase, was added to each well, and the absorbance was measured 1 and 5 minutes after TMB addition using a microplate reader (Infinite MPLEX Monochromator, TECAN, Maennedorf, Switzerland). The assay was performed at room temperature.

[0197] Phagocytic assay. After standing for 30 minutes, neutrophils were divided into 3 × 10⁻¹⁶ cells. 5 Cells were seeded at a density of individual cells / mL and incubated with either albumin, recombinant human albumin (both 15 mg / mL), or a media control in a 5% CO2 incubator at 37°C for 2 hours. Then, 50 μL of opsonized fluorescein conjugate zymosan bioparticles (Thermo Fisher Scientific) were added to each well (cell / bioparticle ratio, 1:10) to a final volume of 200 μL, and incubated at 37°C for 60 minutes. The cells were then washed with sterile DPBS, and 100 μL of trypan blue solution (diluted 1 / 10 with sterile DPBS) was added to quench the fluorescence of the extracellular bioparticles. Finally, the plate was centrifuged at 400 g at room temperature for 5 minutes, and excess trypan blue was carefully aspirated and removed. Fluorescence intensity in each well was measured using a microplate reader (FLUOstar Optima, Ortenberg, Germany).

[0198] The results indicate that albumin can restore the antibacterial function of neutrophils deficient in albumin. Advantageously, administration of human albumin to patients with decompensated cirrhosis may yield clinical benefits such as prevention of infections and suppression of pathogen-associated molecular pattern (PAMP)-induced systemic inflammation.

[0199] The reader's attention is directed to all papers and documents filed in connection with this Application, either concurrently with or prior to this Application, and made available to the public together with this Application, the contents of all such papers and documents are incorporated herein by reference.

[0200] All features disclosed herein (including any appended claims, abstract and drawings), and / or all steps of any method or process disclosed herein, may be combined in any combination, except for any combination in which at least some of such features and / or steps are mutually exclusive.

[0201] Each feature disclosed herein (including any attached claims, abstract, and drawings) may be replaced by an alternative feature serving the same, equivalent, or similar purpose unless otherwise expressly stated. Accordingly, unless otherwise noted, each disclosed feature is merely one example of a general set of equivalent or similar features.

[0202] The present invention is not limited to the details of the embodiments described above. The present invention extends to any novel one or any novel combination of any of the features disclosed herein (including any appended claims, abstract and drawings), or any novel one or any novel combination of any of the steps of any of the methods or processes so disclosed.

[0203] References 1. Moreau R, Jalan R, Gines P, et al. Acute-on-chronic liver failure is a distinct syndrome that develops in patients with acute decompensation of cirrhosis. Gastroenterology 2013;144:1426-37.e1-9 2. Arroyo V, Moreau R, Jalan R. Acute-on-Chronic Liver Failure. N Engl J Med 2020;382:2137‐45. 3. Trebicka J, Fernandez J, Papp M, et al. The PREDICT study uncovers three clinical courses in acutely decompensated cirrhosis with distinct pathophysiology. J Hepatol 2020;73:842-854. 4. Trebicka J, Fernandez J, Papp M, et al. PREDICT identifies precipitating events associated with the clinical course of acutely decompensated cirrhosis. J Hepatol 2021;74:1097-1108. 5. Weiss E, de la Grange P, Defaye M, et al. Characterization of Blood Immune Cells in Patients With Decompensated Cirrhosis Including ACLF. Front Immunol 2021;11:619039.e1-9. 6. Claria J, Stauber RE, Coenraad MJ, et al. Systemic inflammation in decompensated cirrhosis: characterization and role in acute-on-chronic liver failure. Hepatology 2016;64:1249-1264. 7. Lopez-Vicario C, Checa A, Urdangarin A, et al. Targeted lipidomics reveals extensive changes in circulating lipid mediators in patients with acutely decompensated cirrhosis. J Hepatol 2020;73:817-828. 8. O'Brien AJ, Fullerton JN, Massey KA, et al. Immunosuppression in acutely decompensated cirrhosis is mediated by prostaglandin E2. Nat Med 2014;20:518-523. 9. Bernsmeier C, Pop OT, Singanayagam A, et al. Patients with acute-on-chronic liver failure have increased numbers of regulatory immune cells expressing the receptor tyrosine kinase MERTK. Gastroenterology 2015;148:603‐15.e14. 10. Korf H, du Plessis J, van Pelt J, et al. Inhibition of glutamine synthetase in monocytes from patients with acute-on-chronic liver failure resuscitates their antibacterial and inflammatory capacity. Gut 2019;68:1872-1883. 11. Fernandez J, Acevedo J, Wiest R, et al. Bacterial and fungal infections in acute-on-chronic liver failure: prevalence, characteristics and impact on prognosis. Gut 2018;67:1870-80. 12. Bajaj JS, O’Leary JG, Reddy KR, et al. Survival in infection-related acute-on-chronic liver failure is defined by extrahepatic organ failures. Hepatology 2014;60:250-256. 13. European Association for the Study of the Liver. EASL Clinical Practice Guidelines for the management of patients with decompensated cirrhosis. J Hepatol 2018;69:406‐60. 14. Sort P, Navasa M, Arroyo V, et al. Effect of intravenous albumin on renal impairment and mortality in patients with cirrhosis and spontaneous bacterial peritonitis. N Engl J Med 1999;341:403-9. 15. Wong F, Pappas SC, Curry MP, et al. Terlipressin plus Albumin for the Treatment of Type 1 Hepatorenal Syndrome. N Engl J Med 2021;384:818-28. 16. Fernandez J, Claria J, Amoros A, et al. Effects of Albumin Treatment on Systemic and Portal Hemodynamics and Systemic Inflammation in Patients With Decompensated Cirrhosis. Gastroenterology 2019;157:149-62. 17. Casulleras M, Flores-Costa R, Duran-Gueell M, et al. Albumin internalizes and inhibits endosomal TLR signaling in leukocytes from patients with decompensated cirrhosis. Sci Transl Med 2020;12(566):eaax5135. 18. Caraceni P, Riggio O, Angeli P, at al. Long-term albumin administration in decompensated cirrhosis (ANSWER): an open-label randomised trial. Lancet 2018;391(10138):2417-2429. 19. Banchereau R, Hong S, Cantarel B, et al. Personalized Immunomonitoring Uncovers Molecular Networks that Stratify Lupus Patients. Cell 2016;165:551-565. 20. Li S, Rouphael N, Duraisingham S, et al. Molecular signatures of antibody responses derived from a systems biology study of five human vaccines. Nat Immunol 2014;15:195-204. 21. Hagan T, Gerritsen B, Tomalin LE, et al. Transcriptional atlas of the human immune response to 13 vaccines reveals a common predictor of vaccine-induced antibody responses. Nat Immunol 2022;23:1788-1798. 22. Aran D, Looney AP, Liu L, et al. Reference-based analysis of lung single-cell sequencing reveals a transitional profibrotic macrophage. Nat Immunol 2019;20:163-72. 23. Monaco G, Lee B, Xu W, et al. RNA-Seq Signatures Normalized by mRNA Abundance Allow Absolute Deconvolution of Human Immune Cell Types. Cell Rep 2019;26:1627-40.e7. 24. Yaari G, Bolen CR, Thakar J, et al.Quantitative set analysis for gene expression: a method to quantify gene set differential expression including gene-gene correlations. Nucleic Acids Res 2013;41:e170. 25. Gandoura S, Weiss E, Rautou PE, et al. Gene- and exon-expression profiling reveals an extensive LPS-induced response in immune cells in patients with cirrhosis. J Hepatol 2013;58:936-948. 26. Andreatta M, Carmona SJ. UCell: Robust and scalable single-cell gene signature scoring. Comput Struct Biotechnol J 2021;19:3796-3798. 27. Amulic B, Cazalet C, Hayes GL, et al. Neutrophil function: from mechanisms to disease. Annu Rev Immunol. 2012;30:459-89. 28. Bernsmeier C, van der Merwe S, Perianin A. Innate immune cells in cirrhosis. J Hepatol 2020;73:186-201. 29. Arroyo V, Moreau R, Kamath PS, et al. Acute-on-chronic liver failure in cirrhosis. Nat Rev Dis Primers 2016;2:16041. 30. Ip WK, Medzhitov R .Macrophages monitor tissue osmolarity and induce inflammatory response through NLRP3 and NLRC4 inflammasome activation. Nat Commu. 2015;6:6931. 31. Jiang W, Le J, Wang PY, et al. Extracellular Acidity Reprograms Macrophage Metabolism and Innate Responsiveness. J Immunol 2021;206:3021-3031. 32. Murase M, Kawasaki T, Hakozaki R, et al. Intravesicular Acidification Regulates Lipopolysaccharide Inflammation and Tolerance through TLR4 Trafficking. J Immunol 2018;200:2798-2808. 33. Vidarsson G, Stemerding AM, Stapleton NM, et al. FcRn: an IgG receptor on phagocytes with a novel role in phagocytosis. Blood 2006;108:3573-9. 34. Pyzik M, Rath T, Lencer WI, et al. FcRn: The Architect Behind the Immune and Nonimmune Functions of IgG and Albumin. J Immunol 2015;194:4595-603. 35. Zhao R, Dai H, Arias RJ, et al. Direct activation of the proton channel by albumin leads to human sperm capacitation and sustained release of inflammatory mediators by neutrophils. Nat Commun 2021;12:3855.

[0204] Numbered paragraphs Paragraph 1. A composition comprising human albumin for use in the treatment of deficient immune cell function in subjects with systemic inflammatory response syndrome, (a) compositions containing human albumin are intended to treat neutrophil function deficiencies, and human albumin is intended to be administered to the subject in a dose sufficient to increase the antimicrobial function of neutrophils; and / or (b) Compositions containing human albumin are intended for the treatment of deficient CD4+ T cells, and human albumin is intended to be administered to the patient in 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) Compositions containing human albumin are intended to treat dendritic cell function deficiencies, and human albumin is intended to be administered to subjects in doses sufficient to increase the levels of plasmacytoid dendritic cells; and / or (d) Compositions containing human albumin are intended to treat deficiency in monocyte cell function, and human albumin is intended to be administered to subjects in doses sufficient to increase the level of intermediate HAVCR2+ monocytes. composition.

[0205] Paragraph 2. The composition for use described in Paragraph 1(a), wherein the increase in the antimicrobial function of neutrophils is an increase in neutrophil degranulation and / or neutrophil phagocytosis.

[0206] Paragraph 3. A composition for use as described in Paragraph 1(a) or Paragraph 2, wherein human albumin is to be administered to a subject in a dose sufficient to upregulate the expression of a neutrophil gene, or a combination thereof, selected from the group consisting of CD177, OLFM4, PRG2, MPO, BPI, RETN, LCN2, CEACAM8, and MCEMP1.

[0207] Paragraph 4. Compositions for use according to any one of the preceding paragraphs relating to systemic inflammatory response syndromes, which are diseases selected from the group consisting of decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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.

[0208] Paragraph 5. The composition for use according to any one of the preceding paragraphs, wherein the subject is a patient with sepsis or a patient with a hepatic condition.

[0209] Paragraph 6. The composition for use according to any one of the preceding paragraphs, wherein the subject has a liver condition selected from the group consisting of decompensated cirrhosis, the pre-stage of acute exacerbation of chronic liver failure, and acute exacerbation of chronic liver failure.

[0210] Paragraph 7. A composition comprising human albumin for use in the treatment of deficient immune cell function in a subject requiring it, wherein the deficient immune cell function is (a) Deficient neutrophil function; (b) Deficient central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function; (c) Deficient plasmacytoid dendritic cell function; and (d) Deficient intermediate HAVCR2+ monocyte function Selected from the group consisting of, The aforementioned deficiency in immune cell function relates to diseases selected from the group consisting of systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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. composition.

[0211] Paragraph 8. Human albumin is a composition for use according to any one of the preceding paragraphs, intended for administration by a route of administration selected from the group consisting of intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

[0212] Paragraph 9. A composition for use according to any one of the preceding paragraphs, wherein human albumin is to be administered at multiple intervals as part of a multi-dose regimen.

[0213] Paragraph 10. A multi-dose regimen includes multiple divided doses administered at equal intervals of approximately 1 to 30 days; or A multi-dose regimen includes multiple divided doses administered at unequal intervals of approximately 1 to 30 days; or A multi-dose regimen includes two or more doses until the total cumulative dose is reached. Composition for use as described in paragraph 9.

[0214] Paragraph 11. Human albumin is administered as part of a multi-dose regimen in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval; or Human albumin is administered as part of a multi-dose regimen, in individual doses ranging from approximately 20 g / interval to approximately 200 g / interval. Compositions for use as described in paragraph 9 or 10.

[0215] Paragraph 12. A composition for use according to any one of paragraphs 9 to 11, wherein the administration interval is every 15 days or less.

[0216] Paragraph 13. A composition for use according to any one of the preceding paragraphs, wherein human albumin is human plasma-derived albumin or recombinant human albumin.

[0217] Paragraph 14. A composition for use according to any one of the preceding paragraphs, wherein the concentration of human albumin is between 4% and 25% (w / v); or the concentration of human albumin is approximately 20% (w / v).

Claims

1. A composition comprising human albumin for use in regulating immune cells in subjects with systemic inflammatory response syndrome, (a) Compositions containing human albumin are intended to modulate neutrophil function, and human albumin is intended to be administered to subjects in doses sufficient to increase the antimicrobial function of neutrophils; and / or (b) Compositions comprising human albumin are intended to modulate CD4+ T cells, and the human albumin is intended to be administered to a patient in 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) Compositions containing human albumin are intended to modulate dendritic cells, and human albumin is intended to be administered to subjects in doses sufficient to increase the levels of plasmacytoid dendritic cells; and / or (d) Compositions containing human albumin are intended to modulate monocyte cells, and human albumin is administered to subjects in doses sufficient to increase the level of intermediate HAVCR2+ monocytes. composition.

2. The composition for use according to claim 1(a), wherein the increase in the antimicrobial function of neutrophils is an increase in neutrophil degranulation and / or neutrophil phagocytosis.

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

4. A composition for use according to any one of the above claims, relating to a systemic inflammatory response syndrome, which is a disease selected from the group consisting of decompensated cirrhosis, acute liver failure, the pre-stage of acute exacerbation of chronic liver failure, acute exacerbation of 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 claims, wherein the target is a patient with sepsis or a patient with hepatic disease.

6. The composition for use according to any one of the claims, wherein the subject has a liver condition selected from the group consisting of decompensated cirrhosis, the pre-stage of acute exacerbation of chronic liver failure, and acute exacerbation of chronic liver failure.

7. A composition comprising human albumin for treating deficient immune cell function in a subject requiring it, wherein the deficient immune cell function is (a) Deficient neutrophil function; (b) Deficient central memory ITGB1+ CD4+ T cell function and / or activated CD4+ memory T cell function; (c) Deficient plasmacytoid dendritic cell function; and (d) Deficient intermediate HAVCR2+ monocyte function A composition selected from the group consisting of the following.

8. The composition for use according to claim 7, relating to a disease selected from the group consisting of systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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 deficient immune cell function is related to a disease selected from the group consisting of systemic inflammatory response syndrome, decompensated cirrhosis, acute liver failure, pre-acute exacerbation of chronic liver failure, acute exacerbation of 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. A composition for use according to any one of the claims, wherein human albumin is intended for administration by a route of administration selected from the group consisting of intravenous, subcutaneous, intramuscular, intradermal, intraperitoneal, intrapulmonary, intranasal, oral, rectal, and combinations thereof.

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

11. A multi-dose regimen includes multiple divided doses administered at equal intervals of approximately 1 to 30 days; or A multi-dose regimen includes multiple divided doses administered at unequal intervals of approximately 1 to 30 days; or A multi-dose regimen includes two or more doses until the total cumulative dose is reached. The composition for use according to claim 10.

12. Human albumin is intended to be administered as part of a multi-dose regimen, in individual doses ranging from approximately 5 g / interval to approximately 500 g / interval; or Human albumin is intended to be administered as part of a multi-dose regimen, in individual doses ranging from approximately 20 g / interval to approximately 200 g / interval. A composition for use according to claim 10 or 11.

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

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

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

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