Combinatorial treatment

Abstract

Provided are compositions comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the target is a diseased cell or a pathogen, and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. Also provided are compositions comprising a CD16- granulocyte or precursor thereof for therapeutic uses, and antibodies or antigen-binding proteins for binding to an antigen on a target for therapeutic uses. The therapeutic applications include the treatment of cancer, the treatment of infections, and the treatment of autoimmune diseases.

Description

[0001] COMBINATORIAL TREATMENT

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to combinatorial treatments, such as those comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigenbinding protein, as well as compositions comprising the same.

[0004] BACKGROUND

[0005] Host therapeutic immune responses often involve several types of immune cell and play a vital role in the body’s fight against cancer, infections and virtually all other diseases. However, a subject’s native therapeutic immune response is not always enough to eradicate disease. For example, tumours may be adapted to be immunologically “cold” and may create an immunosuppressive tumour microenvironment (TME) that can render native anti-tumour therapeutic immune responses ineffective.

[0006] As a result of the host therapeutic immune response being adapted to be immunosuppressive, immunotherapies, such as antibody-based immunotherapies, can be of limited use. In particular, some antibody-based immunotherapies exert their effects through antibodydependent cellular cytotoxicity (ADCC) whereby the antibody-based therapy binds to a target and also activates an Fc receptor (FcR) on a host immune effector cell to enable the immune effector cell to lyse the target. However, when the host therapeutic immune response has become immunosuppressive, the antibody-based therapy may be unable to activate the host immune effector cell efficiently resulting in sub-optimal therapy.

[0007] Cell immunotherapies can act to amplify the host therapeutic immune response and are therefore being investigated for their clinical efficacy in diseases such as cancer and infections. However, very few cell therapies have been approved for use, and even those that are approved may be of limited utility. For example, conventional leukocyte infusion therapy is an exemplary cell therapy carried out using apheresis for direct transfer of granulocytes (e.g. peripheral neutrophils) from a donor to a recipient to prevent or treat cancer in the recipient. However, peripheral neutrophils have a very limited shelf-life (typically less than 24 hours) and are not amenable to freeze-thawing, making them difficult to store. In addition, apheresis requires approximately 5 (very rare) donors in order to acquire the required cell volume. Thus, conventional leukocyte infusion therapy is not viable as a reliable therapeutic technique. To overcome the problems associated with the limited shelf-life and storage issues associated with leukocyte infusion therapy, stem cell-based (e.g., haematopoietic stem cell-based) immunotherapies are being investigated. However, stem cell-based immunotherapies can be problematic because the stem cells may need to differentiate into more mature cells before they express therapeutic factors. Meanwhile, differentiation of stem cells can give rise to cells having vastly different expression profiles resulting in potentially severe off-target effects or cumbersome purification protocols prior to administration.

[0008] Accordingly, there is a need for improved immunotherapies. The present invention addresses one or more of the above-mentioned problems.

[0009] SUMMARY OF THE INVENTION

[0010] The present inventors have found that a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) can be manufactured that is CD16- and expresses an antibody dependent cellular cytotoxicity (ADCC)-promoting Fc receptor (FcR). Therefore, the invention advantageously provides a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is capable of inducing lysis of a target (e.g., a target cell) through ADCC, and which may be efficacious in promoting ADCC in the treatment of a disorder in a subject.

[0011] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may exhibit several advantages compared to less mature stem cells and native peripheral granulocytes. In particular, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) expresses an ADCC-promoting FcR, which is a class of receptor which aids in lysis of target cells that is not normally expressed in haematopoietic stem cells. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may also be at a differentiation stage where it is committed to a desired lineage, meaning it has limited capacity to give rise to multiple different types of cells and is therefore unlikely to result in off-target effects after administration to a subject. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may thereby overcome many of the disadvantages associated with less mature stem cells. As discussed in more detail below, the granulocyte precursor cell may have been differentiated in vitro to a stage of differentiation corresponding to that of a promyelocyte, a myelocyte, or an intermediate cell thereof. In addition, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16-. CD16b, a subtype of CD16, is very abundant on mature peripheral neutrophils where it acts to decrease ADCC activity of the neutrophils as discussed in Treffers et al Front Immunol. 2019 30:9:3124. Thus, lack of expression of CD16 on a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be particularly beneficial when using the granulocyte or precursor thereof (e.g. , a granulocyte precursor cell) to induce ADCC of a target. The granulocyte or precursorthereof (e.g., a granulocyte precursor cell) may also be generated in large numbers from widely available cells, may be amenable to freeze-thawing and may be cultured after freeze-thawing. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may thereby overcome many of the disadvantages associated with peripheral neutrophils. A granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses the ADCC-promoting FcR CD64 may be particularly advantageous compared to more mature peripheral neutrophils because peripheral neutrophils normally lack CD64 expression. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may otherwise have one or more properties of a peripheral neutrophil.

[0012] The inventors have found that administration of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an ADCC-promoting Fc receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR may provide for improved therapy of a disorder. For example, the combination may result in improved cytotoxicity of a target in a subject compared to administering either constituent alone. In other words, the therapeutic effect associated with the combination may be synergistic. In particular, the combination may induce ADCC of a target.

[0013] DETAILED DESCRIPTION

[0014] In one aspect, the invention provides a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) for use in treating a disorder in a subject in combination with an antibody or antigen-binding protein for binding to an antigen on a target, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In a related aspect, the invention provides a method of treating a disorder in a subject in combination with an antibody or antigen-binding protein for binding to an antigen on a target, the method comprising administering a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) to the subject, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In a related aspect, the invention provides use of a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) in the manufacture of a medicament for treating a disorder in combination with an antibody or antigen-binding protein for binding to an antigen on a target, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD 16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)- promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

[0015] In one aspect, the invention provides an antibody or antigen-binding protein for binding to an antigen on a target for use in treating a disorder in a subject in combination with a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In a related aspect, the invention provides a method of treating a disorder in a subject in combination with a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , the method comprising administering the antibody or antigenbinding protein to a subject, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD 16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)- promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In a related aspect, the invention provides use of an antibody or antigen-binding protein for binding to an antigen on a target in the manufacture of a medicament for treating a disorder in a subject in combination with a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

[0016] In one aspect, the invention provides a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target for use in treating a disorder in a subject, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In a related aspect, the invention provides a method of treating a disorder in a subject, the method comprising administering a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target to a subject, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In a related aspect, the invention provides use of a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target in the manufacture of a medicament for treating a disorder in a subject, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

[0017] In one aspect, the invention provides a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

[0018] In one aspect, the invention provides a composition described herein for use in treating a disorder in a subject. In a related aspect, the invention provides a method of treating a disorder in a subject, the method comprising administering a composition described herein to the subject. In a related aspect, the invention provides use of a composition described herein in the manufacture of a medicament for treating a disorder in a subject.

[0019] In one aspect, the invention provides a kit comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. In one aspect, the invention provides a kit comprising a composition or a pharmaceutical composition described herein.

[0020] In one aspect, the invention provides a method for manufacturing a composition or a kit of the invention, wherein the method comprises combining a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or an antigen-binding protein for binding to an antigen on a target, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

[0021] In another aspect, the invention provides use of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target in a method of manufacturing a composition or a kit of the invention, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD 16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

[0022] A “subject” as used herein may be a mammal. A subject may be a livestock mammal, a domesticated mammal, such as a pet, or a wildlife species. A subject may be a human, a horse, a monkey, a cow (including a bull), a pig, a dog, a cat, a sheep, a goat, an elephant, a panda, a mouse, a rabbit, a rat, or other mammal. Preferably “subject” means a human subject. A “subject” is preferably an adult human subject, i.e. a human subject of at least 18 years old. Alternatively, a subject may be a paediatric human subject less than 18 years old, e.g. at least 1 , 5, 10 or 15 years old but less than 18 years old. The terms “subject” and “patient” are used synonymously herein.

[0023] The term “granulocyte” as used herein may refer to a class of white blood cells characterized by the presence of one or more granules in their cytoplasm. A granulocyte may be a neutrophil, eosinophil, or a basophil. Preferably, the granulocyte may be a neutrophil.

[0024] The term “granulocyte precursor” (e.g. as used in the context of “granulocyte or precursor thereof (e.g., a granulocyte precursor cell)”) may refer to a cell that is capable of differentiating into a granulocyte. Preferably, the granulocyte or granulocyte precursor thereof may be a granulocyte precursor. The granulocyte precursor may be a common myeloid progenitor cell, a granulocyte monocyte progenitor cell, a myeloblast, a promyelocyte, a myelocyte, a metamyelocyte, a band cell, or any intermediate cell thereof or a cell equivalent thereto. An intermediate may exhibit one or more characteristics of the two types of cells described herein to which it is the intermediate. Preferably, the granulocyte precursor may be committed to becoming a granulocyte. The term “granulocytic” is also used to refer to “granulocyte or precursor thereof (e.g., a granulocyte precursor cell)” or “granulocytes or precursors thereof”.

[0025] The granulocyte precursor cell may be a promyelocyte, or a myelocyte, or an intermediate cell thereof. The granulocyte precursor cell may have been differentiated in vitro. The granulocyte precursor cell may have been differentiated in vitro to a stage of differentiation corresponding to that of a promyelocyte, a myelocyte, or an intermediate cell thereof, that has differentiated naturally in vivo.

[0026] The granulocyte precursor cell may have been differentiated in vitro from a stem cell obtained from a donor having neutrophils suitable for treating a disorder. The granulocyte precursor cell may have been differentiated in vitro from a stem cell obtained from a donor having neutrophils suitable for treating cancer and / or an infection.

[0027] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be an HSC or any granulocyte or granulocyte precursor downstream therefrom. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) of the invention (e.g., for use in a method of the invention) may be any granulocyte or precursor thereof (e.g., a granulocyte precursor cell) described herein.

[0028] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be defined with reference to its differentiation state within the granulopoiesis pathway. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a cell that is downstream of a common myeloid progenitor, e.g., a granulocyte monocyte progenitor cell, a myeloblast, a promyelocyte, a myelocyte, a metamyelocyte, or a band cell. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a cell that is downstream of a granulocyte monocyte progenitor cell, e.g., a myeloblast, a promyelocyte, a myelocyte, a metamyelocyte, or a band cell. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a cell that is downstream of a myeloblast, e.g., a promyelocyte, a myelocyte, a metamyelocyte, or a band cell. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a cell that is downstream of a promyelocyte, e.g., a myelocyte, a metamyelocyte, or a band cell.

[0029] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to that of a pre-metamyelocyte, e.g., a myeloblast, a promyelocyte, or a myelocyte. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a cell that is downstream of a myeloblast and that is a pre-metamyelocyte, e.g., a promyelocyte, or a myelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a cell that is downstream of a promyelocyte and that is a pre-metamyelocyte, e.g., a myelocyte.

[0030] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to that from a myeloblast to a band cell. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to that from a myeloblast to a metamyelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to that from a myeloblast to a myelocyte. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to that from a promyelocyte to a myelocyte, e.g., a differentiation stage corresponding to a promyelocyte, or a myelocyte, or any intermediate thereof. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to that from a myeloblast to a promyelocyte.

[0031] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a myeloblast, a promyelocyte, a myelocyte, a metamyelocyte, or a band cell. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a promyelocyte or a myelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a granulocyte, wherein the granulocyte is not a terminally-differentiated granulocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may have a differentiation stage corresponding to a granulocyte, wherein the granulocyte is a terminally-differentiated granulocyte. As set out elsewhere in the specification, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be derived ex vivo from a stem cell, such as a haematopoietic stem cell (HSC) or an iPSC. It will be appreciated that such granulocytes or precursors thereof may not be identical to naturally occurring cells, but may share structural (e.g. marker expression) or functional (e.g. potency) characteristics with such naturally occurring cells. The reference to cells having differentiation stages “corresponding to” named cell types in this disclosure should be interpreted accordingly.

[0032] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be selected from the group comprising (or consisting of): a myeloblast; a promyelocyte; a myelocyte; a metamyelocyte; a band cell; and a granulocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be selected from the group comprising (or consisting of): a myeloblast; a promyelocyte; a myelocyte; a metamyelocyte; and a band cell. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be selected from the group comprising (or consisting of): a myeloblast; a promyelocyte; a myelocyte; and a metamyelocyte. In particular, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be selected from the group comprising (or consisting of): a myeloblast; a promyelocyte; and a myelocyte. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be selected from the group comprising (or consisting of): a promyelocyte and a myelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be selected from the group comprising (or consisting of): a myeloblast; and a promyelocyte.

[0033] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a myeloblast. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a promyelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a myelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a metamyelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a band cell.

[0034] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be committed to the granulocyte lineage. For example, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be committed to the granulocyte lineage and not the monocyte lineage. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be committed to the neutrophil lineage. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a neutrophil or precursor thereof. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a neutrophil precursor cell. Thus, the promyelocyte, myelocyte, metamyelocyte, or band cell described herein may be a neutrophilic promyelocyte, a neutrophilic myelocyte, a neutrophilic metamyelocyte, or a neutrophilic band cell respectively. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a cell selected from the group comprising (or consisting) of: a neutrophilic promyelocyte; a neutrophilic myelocyte; a neutrophilic metamyelocyte; and a neutrophilic band cell. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a neutrophilic promyelocyte or a neutrophilic myelocyte.

[0035] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be defined by reference to being positive or negative for expression of one or more markers. Unless specified otherwise (for example, in lists reciting “or” or “and / or”), references in the present disclosure to cells being positive or negative for expression of a number of specified markers should be taken as requiring the cells in question to have the recited expression (either positive or negative) of each of the markers referred to. Thus, by way of example, reference to a cell as “CD15+ CD66b+” should be taken as meaning that the cell is positive for the expression of both CD15 and CD66b. Methods of determining whether a cell expresses a protein are known in the art, for example by using flow cytometry. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express a protein described herein as determined using flow cytometry.

[0036] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an ADCC-promoting FcR. CD16 expression is generally turned on during granulopoiesis at the metamyelocyte stage. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD16- may be a pre-metamyelocyte, such as a myeloblast, a promyelocyte, or a myelocyte. The CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a myeloblast, a promyelocyte, or a myelocyte. CD16b, a subtype of CD16, is very abundant on peripheral neutrophils where it acts to decrease ADCC of the neutrophil. Thus, lack of expression of CD16 on a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be particularly beneficial in granulocytes or precursors thereof for use in combination with an antibody or antigen-binding protein to induce ADCC of a target.

[0037] As used herein, reference to CD16- may mean CD16a- or CD16b-. Reference to CD16- may mean CD16b-. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16b-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16a+ or CD16a-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16b-; and CD16a+ or CD16a-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16b- and CD16a+. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16b- and CD16a-.

[0038] A granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that expresses an ADCC-promoting FcR may have a differentiation stage corresponding to a cell that is downstream of a myeloblast (e.g., a promyelocyte, a myelocyte, a metamyelocyte, a band cell, or a granulocyte). A granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD16- and expresses an ADCC-promoting FcR may have a differentiation stage corresponding to a cell that is downstream of a myeloblast and that is a pre-metamyelocyte. Thus, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD16- and expresses an ADCC-promoting FcR may have a differentiation stage corresponding to a promyelocyte or a myelocyte, or an intermediate thereof.

[0039] Fc receptors are proteins found on the surface of cells that may contribute to the protective functions of the immune system. In particular, Fc receptors may bind to antibodies or antigenbinding proteins that are attached to their target antigen. Once bound to an antibody or antigenbinding protein, ADCC-promoting Fc receptors may activate the cell they are expressed on, stimulating cytotoxic activity of the cell. Thus, “ADCC-promoting FcR” in this context means any Fc receptor that is capable of promoting cytotoxic activity of the cell (e.g., inducing or increasing cytotoxic activity of the cell). Thus, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) expressing an ADCC-promoting Fc receptor may have increased cytolytic ability when the ADCC-promoting Fc receptor binds to an antibody or antigen-binding protein.

[0040] Fc receptors are classified based on the type of antibody or antigen-binding protein that they recognise. For example, Fc gamma receptors bind IgG; Fc alpha receptors bind IgA; and Fc epsilon receptors bind IgE. Another type of Fc receptor, termed the neonatal Fc receptor, also binds IgG. Accordingly, the ADCC-promoting Fc receptor may be an Fc gamma receptor, an Fc epsilon receptor, an Fc alpha receptor or a neonatal Fc receptor. Preferably, the ADCC- promoting Fc receptor may be an Fc gamma receptor or an Fc alpha receptor. The ADCC- promoting FcR may be an IgG FcR, an IgA FcR, or an IgE FcR. Preferably, the ADCC- promoting FcR may be an IgG FcR or an IgA FcR. The ADCC-promoting Fc receptor may be an ADCC-promoting Fc gamma receptor. The ADCC-promoting Fc gamma receptor may be an FcyRI, or an FcyRII receptor. FcyRI is known as CD64 while FcyRII is known as CD32. Accordingly, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD64 and / or CD32. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) expresses CD64. CD64 is not normally expressed by peripheral neutrophils. Accordingly, granulocytes or precursors thereof that express CD64 may be particularly advantageous in promoting ADCC compared to peripheral neutrophils.

[0041] The ADCC-promoting Fc receptor may be an ADCC-promoting Fc alpha receptor. The Fc alpha receptor may be FcaRI or Fca / pR. Preferably, the Fc alpha receptor may be FcaRI. FcaRI is also known as CD89. Accordingly, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD89. The ADCC-promoting FcR may be CD64, CD32, CD89, or a combination thereof.

[0042] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD64+, CD89+ and / or CD32+. The granulocyte or precursorthereof (e.g., a granulocyte precursor cell) may express more than one ADCC-promoting Fc receptor described herein. Accordingly, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD64 and CD32. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD64 and CD89. The granulocyte or precursorthereof (e.g., a granulocyte precursor cell) may express CD89 and CD32. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD64, CD32 and CD89.

[0043] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD64+, CD32+ and / or CD89+, and (b) CD16-. A granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is (a) CD64+, CD32+ and / or CD89+, and (b) CD16- may express an ADCC-promoting FcR and lack expression of an ADCC-repressing FcR (e.g., CD16b) and may therefore particularly suitable for use in inducing ADCC of a target. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD64+ and CD32+, and (b) CD16-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD64+ and CD89+, and (b) CD16-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD32+ and CD89+, and (b) CD16-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD64+, CD32+ and CD89+, and (b) CD16-. Preferably, the CD16- cells employed in the present invention are granulocytes precursor cells. The granulocyte precursor cells may be: common myeloid progenitor cells, granulocyte monocyte progenitor cells, myeloblasts, promyelocytes, myelocytes, metamyelocytes, band cells, or any intermediate cells thereof or cells equivalent thereto. More preferably, the granulocyte precursor cells are promyelocytes, myelocytes or cells that are an intermediate thereof. Yet more preferably, the granulocyte precursor cells are promyelocytes, myelocytes, or cells that are intermediate thereof or any combination thereof.

[0044] Cells described herein are preferably human cells (e.g. derived from human cells).

[0045] The term “granulocyte” as used herein may refer to a class of white blood cells characterized by the presence of one or more granules in their cytoplasm. A granulocyte may be a neutrophil, eosinophil, or basophils. Preferably, the granulocyte is a neutrophil.

[0046] The term “granulocyte precursor” (e.g. as used in the context of “granulocyte or precursor thereof’) refers to a cell that is capable of differentiating into a granulocyte. A granulocyte precursor may be any granulocyte precursor downstream from an HSC. A granulocyte precursor may be a common myeloid progenitor cell, a granulocyte monocyte progenitor cell, a myeloblast, a promyelocyte, a myelocyte, a metamyelocyte, a band cell, or any intermediate cell thereof or a cell equivalent thereto. Preferably, a granulocyte precursor may be a promyelocyte, a myelocyte, or an intermediate thereof, more preferably an intermediate thereof. An intermediate may exhibit one or more characteristics of the two types of cells described herein to which it is the intermediate. The granulocytes precursor may be a population of cells comprising a plurality of a single granulocyte precursor type, e.g. a population of cells comprising a plurality of myeloblasts or it may be a population of cells comprising a mixture of different granulocytes precursor types, e.g. a population of cells comprising a mixture of common myeloid progenitor cells, granulocyte monocyte progenitor cells, myeloblasts, promyelocytes, myelocytes, metamyelocytes, band cells, and / or any intermediate cells.

[0047] More preferably, the granulocyte precursors may be a population of cells comprising a plurality of a single granulocyte precursor type, e.g. a population of cells comprising a plurality of myeloblasts or a population of promyelocytes or a population of myelocytes or a population of cells that are an intermediate between a promyelocyte and a myelocyte or it may be a population of cells comprising a mixture (e.g. combination) of different granulocytes precursor types, e.g. a population of cells comprising a mixture (e.g. combination) of promyelocytes and myelocytes or a population of cells comprising a mixture of common myeloid progenitor cells, granulocyte monocyte progenitor cells, myeloblasts, promyelocytes, myelocytes, metamyelocytes, band cells, and / or any intermediate cells. Preferably, the granulocyte precursor cell is a promyelocyte, a myelocyte, or a cell that is an intermediate thereof. More preferably, the granulocyte precursor is a combination of promyelocytes, myelocytes, and cells that are intermediate thereof.

[0048] Preferably, a granulocyte precursor may be committed to becoming a granulocyte. A granulocyte precursor is preferably committed to becoming a granulocyte, such as a neutrophil, in vitro, in vivo or ex vivo.

[0049] A granulocyte or precursor thereof according to the present invention may not necessarily be identical to a granulocyte or precursor thereof found in vivo (e.g. in a human). The granulocyte or precursor thereof may have one or more characteristics in common with a corresponding granulocyte or precursor thereof found in vivo (e.g. in a human). The granulocyte or precursor thereof may have one or more characteristics that are different to a corresponding granulocyte or precursor thereof found in vivo (e.g. in a human). Preferably, the granulocyte or precursor thereof may have one or more characteristics in common and one or more characteristics that are different to a corresponding granulocyte or precursor thereof found in vivo (e.g. in a human). A granulocyte or precursor thereof is preferably one that has been differentiated in vitro. The granulocyte or precursor thereof may have one or more characteristics in common with a corresponding granulocyte or precursor thereof produced by a different in vitro method. The granulocyte or precursor thereof may have one or more characteristics that are different to a corresponding granulocyte or precursor thereof produced by a different in vitro method. Preferably, the granulocyte or precursor thereof may have one or more characteristics in common and one or more characteristics that are different to a corresponding granulocyte or precursor thereof produced by a different in vitro method. The granulocyte or precursor thereof is preferably an equivalent granulocyte or precursor thereof. Said one or more characteristics may include one or more of: morphology; cell surface markers; a gene expression profile; cancer killing activity; immunomodulatory properties; persistence; viability; and / or longevity.

[0050] Preferably, a granulocyte or precursor thereof (preferably an in vitro differentiated granulocyte precursor cell) according to the present invention is not identical to an in vivo differentiated natural (wild type) granulocyte or precursor thereof, preferably an in vivo differentiated granulocyte precursor of an equivalent developmental stage, found in vivo (e.g. in a human).

[0051] As exemplified in the examples section, the in vitro differentiated granulocyte precursor cells of the invention may exhibit several distinctive characteristics (e.g. functional and structural), which are not found in granulocyte precursor cells of an equivalent developmental stage that have been differentiated in vivo (e.g. in humans). Without wishing to be bound by theory, it is believed that these unique characteristics, associated with the benefits disclosed herein, are imparted, at least in part, both by the source the start material (HSCs obtained from donors with granulocytes (preferably neutrophils) with high CKA as taught herein via genetically based mechanisms) and by the methods and culture media of the invention.

[0052] The skilled person understands that, when comparing cells, it is important to identify or determine which “comparator” cells to use for the comparison. The skilled person knows that the comparator cell is preferably a cell at a similar or closely matching point in development, and which is derived from the same organism (e.g. human), so that any differences such as a difference in gene expression, cell surface marker expression, cancer killing activity, immunomodulatory activity or other phenotypes reflect true biological differences (e.g. differences attributed to the methods and media of the invention) rather than artifacts such as developmental timing mismatches, organism mismatches, or biases such as artificial genetic manipulations, exposure to compounds, etc.

[0053] The term “equivalent” as used in the context of a granulocyte or granulocyte precursor herein may mean a cell that is from the same organism and / or at an equivalent (preferably identical) developmental stage. Preferably, an equivalent granulocyte or granulocyte precursor is a cell that is from the same organism and at an equivalent (preferably identical) developmental stage. For example, when a human granulocyte precursor cell of the invention that has been differentiated in vitro is an intermediate between a promyelocyte and a myelocyte, an equivalent granulocyte precursor cell that has been differentiated in vivo is preferably a human granulocyte precursor cell is an intermediate between a promyelocyte and a myelocyte that has been differentiated in vivo. Preferably, conversely, a neutrophil isolated from peripheral blood is not equivalent to a granulocyte precursor cell of the invention. It is preferred that an equivalent cell has not been genetically engineered and / or exposed to one or more compounds that are not normally found in vivo.

[0054] The person skilled in the art knows how to determine if two cells are of an equivalent developmental stage. Techniques are known in the art to assign a cell to a particular developmental stage. Such cells may share one or more of characteristics known to be associated with a cell of a particular developmental stage (e.g. in vivo). Said one or more characteristics preferably distinguish said cell from a cell of a different developmental stage. Such characteristics may be one or more of: morphology; cell surface markers; a gene expression profile; cancer killing activity; immunomodulatory properties; persistence; viability; and / or longevity. Preferably, an equivalent developmental stage is the same developmental stage.

[0055] Suitably, a CD16- granulocyte precursor cell has been differentiated in vitro, wherein the granulocyte precursor cell comprises: increased expression of one or more of: serglycin (SRG / V), myeloperoxidase (MPO), major histocompatibility complex, class II, DR alpha (HLA- DRA), CD74, and elastase (ELANE) when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0056] In one aspect, the present invention relates to a granulocyte precursor cell that has been differentiated in vitro, wherein the granulocyte precursor cell may comprise: decreased expression of one or more of: defensin alpha 1 (DEFA1), defensin alpha 3 (DEFA3), cathelicidin antimicrobial peptide (CAMP), bactericidal permeability increasing protein (BPI), and azurocidin 1 (AZU1) when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0057] In one aspect, the present invention relates to a CD16- granulocyte precursor cell that has been differentiated in vitro, wherein the granulocyte precursor cell may comprise: decreased expression of one or more of: defensin alpha 1 (DEFA1), defensin alpha 3 (DEFA3), cathelicidin antimicrobial peptide (CAMP), bactericidal permeability increasing protein (BPI), and azurocidin 1 (AZU1) when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0058] In a further aspect, the present invention relates to a granulocyte precursor cell that has been differentiated in vitro, wherein the granulocyte precursor cell comprises: i. increased expression of one or more of: SRGN, MPO, HLA-DRA, CD74, and ELANE when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo, and / or ii. decreased expression of one or more of: DEFA1, DEFA3, CAMP, BPI, and AZU1 when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0059] In a further aspect, the present invention relates to a granulocyte precursor cell that has been differentiated in vitro, wherein the granulocyte precursor cell is CD16- and comprises: iii. increased expression of one or more of: SRGN, MPO, HLA-DRA, CD74, and ELANE when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo, and / or iv. decreased expression of one or more of: DEFA1, DEFA3, CAMP, BPI, and AZU1 when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0060] In a preferred aspect, the present invention relates to a granulocyte precursor cell that has been differentiated in vitro, wherein the granulocyte precursor cell comprises: a. increased expression of one or more of: SRGN, MPO, HLA-DRA, CD74, and ELANE when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo, and b. decreased expression of one or more of: DEFA1, DEFA3, CAMP, BPI, and AZU1 when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0061] In a preferred aspect, the present invention relates to a granulocyte precursor cell that has been differentiated in vitro, wherein the granulocyte precursor cell is CD16- and comprises: a. increased expression of one or more of: SRGN, MPO, HLA-DRA, CD74, and ELANE when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo, and b. decreased expression of one or more of: DEFA1, DEFA3, CAMP, BPI, and AZU1 when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0062] The term “increased expression” or “decreased expression” may mean increased and / or decreased expression of least 1 gene such as 1 , 2, 3, 4, or 5, preferably at least 2 genes, preferably at least 3 genes, more preferably at least 4 genes, yet more preferably at least 5 genes. Most preferably, a granulocyte precursor cell may comprise increased expression of all of the listed genes and / or (preferably and) decreased expression of all the listed genes when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

[0063] The granulocyte precursor cell that has been differentiated in vitro from a stem cell may be a granulocyte precursor cell that has been differentiated in vitro from any suitable stem cell. Preferably, the granulocyte precursor cell has been differentiated in vitro from a stem cell that has been derived or has been obtained from a (human) donor having granulocytes (neutrophils) with high cancer killing activity (CKA).

[0064] The stem cell may be any suitable stem cells from the donor, such as HSCs and iPSC. Preferably the stem cells is an HSC. Preferably, the granulocyte precursor cell has been differentiated in vitro from an HSC that has been derived or has been obtained from a (human) donor having granulocytes (neutrophils) with high cancer killing activity (CKA).

[0065] The CKA of a (preferably human) granulocytes (preferably neutrophils) obtained from a donor may be determined or measured by any suitable method in vitro).

[0066] Preferably, the CKA of the (preferably human) granulocytes (preferably neutrophils) is assessed by a method comprising: a. admixing granulocytes (preferably neutrophils) from a donor with cancer cells; b. incubating said admixture; and c. measuring the % of cancer cells killed in said admixture, thereby assessing the CKA of said granulocytes (neutrophils).

[0067] Measuring the % of cancer cells killed in step (c) may be performed as taught herein.

[0068] A granulocyte (preferably a neutrophil) with high CKA may be a granulocyte (preferably a neutrophil) having a CKA of at least 30%, for instance at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or more. Preferably, a granulocyte (preferably a neutrophil) with high CKA is a granulocyte (preferably a neutrophil) having a CKA of at least 30%, preferably at least 50%, more preferably at least 70%.

[0069] The equivalent granulocyte precursor cell that has been differentiated in vivo may be obtained from peripheral blood or from a cell bank. Preferably, the equivalent granulocyte precursor cell that has been differentiated in vivo is a peripheral blood derived granulocyte precursor cell.

[0070] Preferably, the increased expression and / or decreased expression in one or more of the genes of the invention is not the result of a genetic modification of the granulocyte precursor cells (preferably promyelocytes, myelocytes and / or intermediate cells thereof). The term “serglycin (SRGN)” as used herein refers to a protein encoded by the SRGN gene in humans, known as a hematopoietic proteoglycan core protein or secretory granule proteoglycan core protein. SRGN typically functions as a scaffold protein in secretory granules of various immune cells, where it often helps in storing, secreting, and protecting proteases, chemokines, and other molecules. SRGN may be associated with the macromolecular complex of granzymes and perforin, which may serve as a mediator of granule-mediated apoptosis. The present inventors surprisingly found that SRGN was highly expressed in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because SRGN expression level was much higher than what is typically observed in natural (wild-type) cells (e.g. from human blood) of an equivalent developmental stage that have been differentiated in vivo or a cell of an equivalent developmental stage differentiated in vitro and obtained from a different method as evidenced for instance in: Korpetinou et al (2014) Frontiers in Oncology: Vol 3, Article 327, pages 1-12, available at: https:.'7ww'w.frontiersin.ofq / iournals / oncoiogy / articies / 10.3389 / fonc.2013.00327 / fuil). Without wishing to be bound by any theories, it is believed that such (relatively) high level of SRGN expression is indicative of cells (preferably granulocyte precursor cells) that are functionally primed as effector cells (e.g. activated cells capable of carrying out a specific response to eliminate threats like infections or abnormal cells such as cancer cells) enabling them to initiate cytotoxic or immunomodulatory responses at an earlier stage of maturation than typically observed.

[0071] The term “myeloperoxidase (MPO)” as used herein refers to a heme-containing enzyme located in the azurophilic (primary) granules of neutrophils, where it typically contributes to antimicrobial defence by catalysing the production of reactive substances such as hypochlorous acid from hydrogen peroxide and chloride ions. Additionally, MPO typically plays a role in mediating cytotoxic activity, including the killing of abnormal cells like cancer cells, through the generation of potent reactive oxygen species (ROS). The present inventors surprisingly found that MPO was highly expressed in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because MOP expression level was much higher than what is typically observed in natural (wild-type cells, e.g. from human blood) of an equivalent developmental stage that have been differentiated in vivo or a cell of an equivalent developmental stage that have been differentiated in vitro and obtained from a different method as evidenced in e.g. Rizo-Tellez et al (2022) Vol 1 (11), 2302, available at: https: / / www.mdpi.eom / 2076-3921 / 11 / 11 / 2302). Without wishing to be bound by any theories, it is believed that the relatively high expression level of MPO is indicative of cells (preferably granulocyte precursor cells) equipped with antimicrobial and cytotoxic functions, thereby possessing an elevated cytocidal potential at an earlier stage of maturation than is typically observed.

[0072] The term “major histocompatibility complex, class II, DR alpha (HLA-DRA)” as used herein refers to a major histocompatibility complex class II molecule. HLA-DRA is typically expressed on the surface of antigen-presenting cells, such as B lymphocytes, dendritic cells, and macrophages, and plays a critical role in immune function by presenting extracellularly derived peptide antigens to CD4+ T lymphocytes. The present inventors surprisingly found that HLA- DRA was highly expressed in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because HLA-DRA is not typically expressed in natural (wild type) granulocytes that have been differentiated in vivo, particularly it is not typically expressed in in vivo differentiated granulocyte precursors. Therefore, said high level of expression of HLA-DRA are unexpected, as evidenced in e.g. McKenna et al in: Frontiers in immunology (2021), Vol 12, Article 602963, available at: https: / Aww.frontiersin.orQ / iournals / mmunoloqy / art!Cies / 10.3389 / fim :j.2021 .602963 / JH).

[0073] Without wishing to be bound by any theories, it is believed that such (relatively) high level of HLA-DRA expression is indicative of cells (preferably granulocyte precursor cells) that have acquired antigen-presenting cell-like properties, which is not a naturally-occurring phenomenon but rather is a property that is (likely) imparted to the cells of the invention by the methods and culture medium of the invention.

[0074] The term “CD74” as used herein refers to a type II transmembrane glycoprotein commonly known as the invariant chain, which typically plays a critical role in the regulation of major histocompatibility complex class II (MHC II) molecule processing and trafficking within antigenpresenting cells. CD74 is typically expressed in antigen-presenting cells such as B cells, monocytes, macrophages, and dendritic cells. The present inventors surprisingly found that CD74 was highly expressed in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because CD74 is not typically expressed in in vivo differentiated granulocytes (typically not in in vitro differentiated granulocyte precursors), and when present on granulocytes (e.g. granulocyte precursor cells), its expression is generally very low or negligible, as evidenced for instance in the “Human Atlas” available at: https: / 7www. proteinatlas. orq / ENSG00000019582-

[0075] CD74?utm source- . Without wishing to be bound by any theories, it is believed that such (relatively) high level of CD74 expression is indicative of cells (preferably granulocyte precursor cells) that have acquired antigen-presenting cell-like properties, which is not a naturally-occurring phenomenon but rather is a property that is (likely) imparted to the cells of the invention by the methods and culture medium of the invention.

[0076] The term “elastase (ELANE)” as used herein refers to a serine protease enzyme typically expressed in neutrophils, where it is stored in azurophil granules. ELANE typically plays a key role in innate immunity by degrading extracellular matrix proteins and microbial components, aiding in pathogen destruction. Additionally, ELANE exhibits selective cytotoxic activity against abnormal cells, including cancer cells, by inducing apoptosis. The present inventors surprisingly found that ELANE was highly expressed in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because ELANE expression level was much higher than what is typically observed in natural (wild-type cells, e.g. from human blood) of an equivalent developmental stage that have been differentiated in vivo or a cell of an equivalent developmental stage that have been differentiated in vitro and obtained from a different method. Without wishing to be bound by any theories, it is believed that the relatively high expression level of ELANE is indicative of cells (preferably granulocyte precursor cells) equipped with antimicrobial and cytotoxic functions, thereby possessing an elevated cytocidal potential at an earlier stage of maturation than is typically observed.

[0077] The term “defensin alpha 1 (DEFA1)” as used herein refers to a small cationic antimicrobial peptide encoded by the DEFA1 gene and typically stored in the azurophil granules of neutrophils. DEFA1 belongs to the alpha-defensin family and exhibits broad-spectrum antimicrobial activities against bacteria, fungi, and viruses by disrupting pathogen membranes. In addition to its antimicrobial function, DEFA1 typically plays a role in cancer killing by selectively targeting and inducing apoptosis in tumour cells, thereby contributing to innate immune defence and tumour suppression. The present inventors found that DEFA1 was expressed at a (very) low level (or undetectable level) in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because DEFA1 expression level was much lower (was undetectable) than what is typically observed in (natural) wild-type cells (e.g. from human blood) of a comparable developmental stage differentiated in vivo or a cell of an equivalent developmental stage differentiated in vitro, obtained from a different method, as evidenced for instance in: Grassi et al in: Cell Reports (2018) Vol 24, pages 2784-2794, available at: Without wishing to be bound by any theories, although the significance of low expression levels of DEFA1 is unknown, it is believed that such (relatively) low level of DEFA1 expression is indicative of cells that have acquired a distinct phenotype and / or gene marker profile that is not naturally occurring in cells of an equivalent developmental stage.

[0078] The term “defensin alpha 3 (DEFA3)” as used herein refers to a small cationic antimicrobial peptide encoded by the DEFA3 gene and typically stored in the azurophil granules of neutrophils. DEFA3, also known as human neutrophil peptide 3 (HNP-3), is a member of the alpha-defensin family, differing from DEFA1 by a single amino acid. DEFA3 typically exhibits antimicrobial activity against bacteria, fungi, and viruses by disrupting microbial membranes, thus often playing a role in innate host defence. Similar to other alpha-defensins, DEFA3 may also contribute to immune modulation and cytotoxicity, supporting the elimination of pathogens and abnormal cells, such as cancer cells. The present inventors found that DEFA3 was expressed at a (very) low level (or undetectable level) in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because DEFA3 expression level was much lower (was undetectable) than what is typically observed in a natural cell (a wild-type cell, e.g. from human blood) a cell (preferably a granulocyte precursor cell) of an equivalent developmental stage that has been differentiated in vivo or a cell an equivalent developmental stage that has been differentiated in vitro and obtained from a different method, as evidenced for instance in: Grassi et al in: Cell

[0079] Reports (2018) Vol 24, pages 2784-2794, available at: Without wishing to be bound by any theories, although the significance of low expression levels of DEFA3 is unknown, it is believed that such (relatively) low level of DEFA3 expression is indicative of cells that have acquired a distinct phenotype and / or gene marker profile that is not naturally occurring in cells of an equivalent developmental stage.

[0080] The term “cathelicidin antimicrobial peptide (CAMP)” as used herein refers to a family of cationic antimicrobial peptides, including the human peptide LL-37, characterised by a conserved cathelin pro-domain and a diverse antimicrobial domain. Specifically, the CAMP gene encodes a precursor protein (hCAP18) that is processed to the active peptide LL-37. CAMP peptides are typically stored in granules of neutrophils and other immune cells and typically display broad-spectrum antimicrobial activity through membrane disruption and intracellular targeting of bacteria, fungi, and viruses. Beyond antimicrobial defence, CAMP peptides, such as LL-37, may also exhibit selective anticancer activity by inducing tumour cell death via membrane permeabilisation and modulation of immune responses. These peptides may also promote wound healing and inflammation regulation, making them multifunctional effectors of innate immunity with therapeutic potential in infectious diseases and cancer. The present inventors found that CAMP was expressed at a (very) low level (or undetectable level) in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because CAMP expression level was much lower (was undetectable) than what is typically observed in a natural cell (a wildtype cell e.g. from human blood) of an equivalent developmental stage that has been differentiated in vivo or a cell of an equivalent developmental stage differentiated in vitro and obtained from a different method, as evidenced for instance in: Nagaoka et al in: Journal of Leukocyte Biology (1998), Vol 64, Issue 6, pages 845-852, available at: https: / / academic.oup.com / ileukbio / article- ab$tract / 64 / '6 / 845 / '6977091?red:rectedFrair;-fulitext). Without wishing to be bound by any theories, although the significance of low expression levels of CAMP is unknown, it is believed that such (relatively) low level of CAMP expression is indicative of cells that have acquired a distinct phenotype and / or gene marker profile that is not naturally occurring in cells of an equivalent developmental stage.

[0081] The term “bactericidal permeability increasing protein (BPI)” as used herein refers to a neutrophil-derived, cationic protein encoded by the BPI gene, which typically plays a role in innate immunity by exhibiting potent antimicrobial activity against gram-negative bacteria. BPI typically binds to lipopolysaccharides in bacterial outer membranes, disrupting membrane integrity and causing bacterial death, while also acting as an opsonin to enhance phagocytosis and antigen presentation. In addition to its antimicrobial functions, BPI often contributes to anticancer activity by modulating immune responses and promoting tumour cell clearance. Thus, BPI is a multifunctional effector that may be involved in direct bacterial killing and immune system-mediated tumour suppression. The present inventors found that BPI was expressed at a (very) low level (or undetectable level) in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because BPI expression level was much lower (was undetectable) than what is typically observed in wild-type cells (e.g. from human blood) of a comparable developmental stage or a cell of a comparable developmental stage obtained from a different method, as evidenced for instance in: Lennartsson et al in: Journal of Leukocyte Biology (2006), Vol 80,

[0082] Issue 1 , pages 196-203, available at: https: / / academic.oup.com / ileukbio / article- Without wishing to be bound by any theories, although the significance of low expression levels of BPI is unknown, it is believed that such (relatively) low level of BPI expression is indicative of cells that have acquired a distinct phenotype and / or gene marker profile that is not naturally occurring in cells of an equivalent developmental stage.

[0083] The term “azurocidin 1 (AZU1)” as used herein refers to a neutrophil-derived cationic glycoprotein encoded by the AZU1 gene that is typically stored in azurophil granules and secretory vesicles. AZU1 typically exhibits potent antimicrobial activity, particularly against gram-negative bacteria, through binding to lipopolysaccharides and disrupting bacterial membranes. Additionally, AZU1 may function as a multifunctional inflammatory mediator and chemotactic factor for monocytes and fibroblasts, promoting immune cell recruitment and vascular permeability. AZU1 may also exhibit anticancer activity. The present inventors found that AZU1 was expressed at a (very) low level (or undetectable level) in the cells of the invention, particularly in granulocyte precursors including promyelocytes, myelocytes or intermediate cells thereof. This is surprising because AZU1 expression level was much lower (was undetectable) than what is typically observed in a natural cell (e.g. a wild-type cell, e.g. from human blood) of an equivalent developmental stage that has been differentiated in vivo or a cell of an equivalent developmental stage that has been differentiated in vitro and obtained from a different method. Without wishing to be bound by any theories, although the significance of low expression levels of AZU1 is unknown, it is believed that such (relatively) low level of AZU1 expression is indicative of cells that have acquired a distinct phenotype and / or gene marker profile that is not naturally occurring in cells of an equivalent developmental stage.

[0084] The genes of the invention (e.g. SRGN, MPO, HLA-DRA, CD74, ELANE, DEFA1, DEFA3, CAMP, BPI, and AZU1) have the following National Center for Biotechnology Information (NCBI) identification (ID) numbers as well as Ensembl_Gene_ID numbers. The skilled person knows how to determine a NCBI ID number and an Ensembl_Gene_ID number for a given gene by, for instance, searching the NCBI gene database at: https: / / www.ncbi.nlm.nih.gov / gene) and Ensembl_Gene_ID database at: https: / / www.ensembl.org.

[0085] Table X: NCBI Gene ID Numbers

[0086] The gene version relating to the NCBI Gene ID Numbers and Ensembl_Gene_ID listed in the Table represents the latest available as of 19 September 2025 for both.

[0087] The granulocyte precursor cell may be one of: a common myeloid progenitor cell, a myeloblast, a neutrophil promyelocyte, a neutrophil myelocyte, a neutrophil metamyelocyte, a neutrophil band, an intermediate cell thereof or any combinations thereof.

[0088] The granulocyte precursor cell may be a promyelocyte, a myelocyte, or an intermediate thereof.

[0089] Preferably, the granulocyte precursor cell of the invention (differentiated in vitro) is a promyelocyte, a myelocyte, or an intermediate thereof.

[0090] Preferably, the granulocyte precursor cell of the invention (differentiated in vitro) is a promyelocyte, a myelocyte, or an intermediate thereof or any combinations thereof.

[0091] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may further express CD66b, CD15, CXCR2, CXCR4, CD18, CD24, CD11 b, CD11c, CD177, CD35, HLA-DR, CD34, CD49d, neutrophil elastase, and / or FasL. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may further express CD15, CXCR2, CXCR4, CD18, CD24, CD11 b, CD11c, CD177, CD10, CD35, HLA-DR, CD34, CD49d, neutrophil elastase, and FasL.

[0092] In some instances, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not express CD10. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16- and CD10-. CD10 expression is generally turned on during granulopoiesis at the granulocyte stage. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD10- may be a pre-granulocyte, such as a myeloblast, a promyelocyte, a myelocyte, a metamyelocyte, or a band cell.

[0093] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may further express CXCR2, CD14, CD19, CD3, CD56, CD68, CD66b, CD15, CXCR2, CXCR4, CD18, CD24, CD11 b, CD11c, CD177, CD35, HLA-DR, CD34, CD49d, neutrophil elastase, and / or FasL, and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not express CD10. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CXCR2+, CD14+, CD19+, CD3+, CD56+, CD68+, CD66b+, CD15+, CXCR2+, CXCR4+, CD18+, CD24+, CD11 b+, CD11c+, CD177+, CD35+, HLA-DR+, CD34+, CD49d+, neutrophil elastase+, and / or FasL+ and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD10-. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may further express CXCR2, CD14, CD19, CD3, CD56, CD68, CD66b, CD15, CXCR2, CXCR4, CD18, CD24, CD11 b, CD11c, CD177, CD35, HLA-DR, CD34, CD49d, neutrophil elastase, and FasL, and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not express CD10. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CXCR2+, CD14+, CD19+, CD3+, CD56+, CD68+, CD66b+, CD15+, CXCR2+, CXCR4+, CD18+, CD24+, CD11 b+, CD11C+, CD177+, CD35+, HLA-DR+, CD34+, CD49d+, neutrophil elastase+, ADCC-promoting FcR+ (e.g., CD64+, CD32+ and CD89+), and FasL+ and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16- and CD10-.

[0094] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD66b+. CD66b expression is generally turned on during granulopoiesis at the promyelocyte stage. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD66b+ may be downstream of a myeloblast.

[0095] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD15+, CD33+, CXCR2+, CXCR4+ and / or CD18+. CD15+, CD33+, CXCR2+, CXCR4+ and CD18+ expression is generally found in all granulocytes or precursors thereof from a myeloblast to a granulocyte (e.g., neutrophil). Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD15+, CD33+, CXCR2+, CXCR4+ and / or CD18+ may be any granulocyte or precursor thereof (e.g., a granulocyte precursor cell) from a myeloblast to a granulocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD15+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD33+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CXCR2+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CXCR4+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD18+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD15+, CD33+, CXCR2+, CXCR4+ and CD18+.

[0096] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD24+, CD177+ CD11 b+ and / or CD11c+. CD24, CD177, CD11 b, and CD11c expression is generally turned on during granulopoiesis at the myelocyte stage. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD24+, CD177+, CD11 b+ and / or CD11c+ may be downstream of a promyelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD24+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD177+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD11 b+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD11c+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD24+, CD177+ CD11 b+ and CD11c+.

[0097] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD35+. CD35 expression is generally turned on during granulopoiesis at the band cell stage. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD35+ may be downstream of a metamyelocyte (e.g., a band cell or a granulocyte).

[0098] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be HLA-DR and / or CD34+. HLA-DR and CD34 expression is generally turned off after the myeloblast stage. Thus, a granulocyte or precursor thereof (e.g. , a granulocyte precursor cell) that is HLA- DR+ and / or CD34+ may have a differentiation stage that corresponds to a pre-promyelocyte e.g., a myeloblast. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be HLA-DR+. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD34+.

[0099] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD49d+. CD49d expression is generally turned off at the band cell stage. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD49d+ may have a differentiation stage that corresponds to a pre-band cell, e.g., a myeloblast, a promyelocyte, a myelocyte, or a metamyelocyte. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express a marker associated with lytic ability of neutrophils (e.g., neutrophil elastase). Such a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be particularly suitable for use in inducing ADCC of a target. Preferably, the granulocyte or precursorthereof (e.g., a granulocyte precursor cell) may further express neutrophil elastase. Neutrophil elastase is secreted by neutrophils during inflammation and destroys bacteria and host tissue. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase may be particularly suitable for lysing targets. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express an ADCC-promoting FcR and neutrophil elastase. Neutrophil elastase is involved in the lysis of targets. Accordingly, a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that expresses an ADCC- promoting FcR and neutrophil elastase may be particularly suitable for use in inducing ADCC of a target.

[0100] Preferably, the granulocyte or precursorthereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) express an ADCC-promoting FcR, and (c) express a marker associated with lytic ability of neutrophils. A granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is (a) CD16-, (b) expresses an ADCC-promoting FcR, and (c) expresses a marker associated with lytic ability of neutrophils may be particularly suitable for use in inducing ADCC of a target. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) express an ADCC-promoting FcR, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD64+, CD32+ and / or CD89+, and (c) express neutrophil elastase. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD64+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD32+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD89+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD64+ and CD32+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16- (b) CD64+ and CD89+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD32+ and CD89+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be (a) CD16-, (b) CD64+, CD32+ and CD89+, and (c) express neutrophil elastase. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD62L-. Lack of expression of CD62L- contrasts to neutrophils found in circulation and at the times of homeostasis. Lack of expression of CD62L may therefore provide a useful means by which the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be distinguished from those that occur naturally. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be CD16-, CD62L- and express an ADCC-promoting FcR.

[0101] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) :

[0102] • may be “Lin-“ (which is to say negative for a cocktail of common leukocyte lineage markers, defined for the present purposes as negative for expression of each of CD3, CD16, CD19, CD20, CD14 and CD56); may be CD38+;

[0103] • may have an HSC phenotype (defined for the present purposes as Lin-CD34+CD38- CD45RA-CD90+);

[0104] • may have a long-term repopulation haematopoietic stem cell (LT-HSC) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-CD90+CD49f+);

[0105] • may have a lymphoid primed multi potent progenitor (LMPP) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA+); and / or

[0106] • may have a multipotent progenitor (MPP) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-).

[0107] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) :

[0108] • may be “Lin-“ (which is to say negative for a cocktail of common leukocyte lineage markers, defined for the present purposes as negative for expression of each of CD3, CD16, CD19, CD20, CD14 and CD56); may be CD38+;

[0109] • may have an HSC phenotype (defined for the present purposes as Lin-CD34+CD38- CD45RA-CD90+);

[0110] • may have a long-term repopulation haematopoietic stem cell (LT-HSC) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-CD90+CD49f+);

[0111] • may have a lymphoid primed multi potent progenitor (LMPP) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA+); and

[0112] • may have a multipotent progenitor (MPP) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-).

[0113] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) described herein may be present in a composition. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) described herein may be part of a composition. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) described herein may be present in a composition in a therapeutically effective amount. Such compositions are also suitable for use with an antibody or antigen-binding protein in the treatment of a disorder. Accordingly, references to a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) herein are also references to a composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) (e.g., a composition comprise a therapeutically effective amount of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ).

[0114] The composition comprises a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and expresses an ADCC-promoting FcR. The composition may comprise a therapeutically effective amount of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and which expresses an ADCC-promoting FcR. At least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD16- and express an ADCC-promoting FcR. Preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD16- and express an ADCC-promoting FcR. Particularly preferably, at least 75% of the granulocytes or precursors thereof in the composition may be CD16- and express an ADCC-promoting FcR.

[0115] The composition comprises a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16-. At least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD16-. Preferably, at least 90% of the granulocytes or precursors thereof in the composition may be CD16-. At least 95% of the granulocytes or precursors thereof in the composition may be CD16-. A composition of the invention may therefore comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD16+. It is preferred that, when present, such granulocytes or precursors thereof represent a low percentage of the total number of cells present (e.g., granulocytes or precursors thereof present). For example, it is preferred that such granulocytes or precursors thereof represent less than 15%, 10% or 5% (preferably less than 1%) of the total number of cells present (e.g., granulocytes or precursors thereof present).

[0116] The composition comprises a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses an ADCC-promoting FcR. At least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may express an ADCC- promoting FcR. Preferably, at least 60% of the granulocytes or precursors thereof in the composition may express an ADCC-promoting FcR. Particularly preferably, at least 75% of the granulocytes or precursors thereof in the composition may express an ADCC-promoting FcR.

[0117] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD64+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD64+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD64+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD64+.

[0118] Preferably, the composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD64+. The composition may comprise a therapeutically effective amount of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD64+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99% or 100% of the granulocytes or precursors thereof in the composition may be CD16- and CD64+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD16- and CD64+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD16- and CD64+.

[0119] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD89+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD89+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD89+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD89+.

[0120] Preferably, the composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD89+. The composition may comprise a therapeutically effective amount of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD89+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99% or 100% of the granulocytes or precursors thereof in the composition may be CD16- and CD89+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD16- and CD89+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD16- and CD89+.

[0121] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD64+, and a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD89+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD 16- and CD64+, and at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD16- and CD89+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD16- and CD64+ and at least 40% of the granulocytes or precursors thereof in the composition may be CD16- and CD89+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD16- and CD64+ and at least 60% of the granulocytes or precursors thereof in the composition may be CD16- and CD89+.

[0122] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD32+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, or 100% of the granulocytes or precursors thereof in the composition may be CD32+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD32+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD32+.

[0123] Preferably, the composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD32+. The composition may comprise a therapeutically effective amount of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16- and CD32+. At least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99% or 100% of the granulocytes or precursors thereof in the composition may be CD16- and CD32+. Preferably, at least 40% of the granulocytes or precursors thereof in the composition may be CD16- and CD32+. Particularly preferably, at least 60% of the granulocytes or precursors thereof in the composition may be CD16- and CD32+. The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is not CD16-, and / or which does not express an ADCC-promoting FcR. The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is not CD16-. The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is CD16+. The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which does not express an ADCC-promoting FcR.

[0124] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which is “Lin-“ (which is to say negative for a cocktail of common leukocyte lineage markers, defined for the present purposes as negative for expression of each of CD3, CD16, CD19, CD20, CD14 and CD56). At least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% of the cells in the composition may be Lin-. Preferably, approximately 95-99% of the cells in the composition may be Lin-, e.g., approximately 97% of the cells in the composition may be Lin- cells.

[0125] The composition may comprise a CD34+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Less than 50%, less than 45%, less than 40%, or less than 35% of the in the composition may be CD34+. By way of example, less than 30% of the cells in the composition may be CD34+. Preferably, approximately 5-25% of the cells in the composition may be CD34+, e.g., approximately 14% of the cells in the composition may be CD34+.

[0126] The composition may comprise a CD38+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . At least 10%, at least 15%, or at least 20% of the cells in the composition may be CD38+. By way of example, between approximately 10% and 80% of the cells in the composition, such as between approximately 10% and 30% of the cells in the composition may be CD38+. Approximately 12% of the cells in the composition may be CD38+.

[0127] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) with a haematopoietic stem cell (HSC) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-CD90+). Less than 5%, less than 4%, less than 3%, or less than 2% of the granulocytes or precursors thereof in the composition may have an HSC phenotype. By way of example, less than 1 % of the granulocytes or precursors thereof in the composition may have an HSC phenotype. Approximately 0.01-0.15% of the granulocytes or precursors thereof in the composition may have an HSC phenotype. Approximately 0.04% of the granulocytes or precursors thereof in the composition may have an HSC phenotype.

[0128] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) having a long-term repopulation haematopoietic stem cell (LT-HSC) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-CD90+CD49f+). Less than 5%, less than 4%, less than 3%, or less than 2% of the cells in the composition may have an LT-HSC phenotype. Byway of example, less than 1% of the cells in the composition may have an LT-HSC phenotype. Approximately 0.01-0.05% of the cells in the composition may have an LT-HSC phenotype. Approximately 0.02% of the cells in the composition may have an LT- HSC phenotype.

[0129] The composition may comprise a cell with a lymphoid primed multi potent progenitor (LMPP) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA+). For example, less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, or less than 25% of the cells in the composition may have an LMPP phenotype. By way of example, less than 20% of the cells in the composition may have an LMPP phenotype. Approximately 2-15% of the cells in the composition may have an LMPP phenotype. Approximately 5% of the cells in the composition may have an LMPP phenotype.

[0130] The composition may comprise a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) with a multipotent progenitor (MPP) phenotype (defined for the present purposes as Lin-CD34+CD38-CD45RA-). Less than 30%, less than 25%, less than 20%, or less than 15% of the cells in the composition may have an MPP phenotype. By way of example, less than 10% of the cells in the composition may have an MPP phenotype. Approximately 1- 6% of the cells in the composition may have an MPP phenotype. Approximately 2% of the cells in the composition may have an MPP phenotype.

[0131] The composition may comprise more than one type of granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Thus, the composition may comprise granulocytes or precursors thereof.

[0132] The composition may comprise (a) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a first ADCC-promoting FcR; (b) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a second ADCC-promoting FcR; and / or (c) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a third ADCC-promoting FcR. The composition may comprise (a) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a first ADCC-promoting FcR; (b) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a second ADCC-promoting FcR; and (c) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a third ADCC-promoting FcR. The first, second and third ADCC-promoting FcR may be different types of ADCC-promoting FcR.

[0133] For example, the composition may comprise (a) a CD16- and CD64+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , (b) a CD16- and CD89+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ; and / or (c) a CD16- and CD32+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The composition may comprise (a) a CD16- and CD64+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , (b) a CD16- and CD89+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ; and (c) a CD16- and CD32+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0134] The composition may comprise (a) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a first ADCC-promoting FcR and neutrophil elastase; (b) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a second ADCC-promoting FcR and neutrophil elastase; and / or (c) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a third ADCC-promoting FcR and neutrophil elastase. The composition may comprise (a) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a first ADCC-promoting FcR and neutrophil elastase; (b) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a second ADCC-promoting FcR and neutrophil elastase; and (c) a CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses a third ADCC-promoting FcR and neutrophil elastase.

[0135] For example, the composition may comprise (a) a CD16- and CD64+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase; (b) a CD16- and CD89+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase; and / or (c) a CD16- and CD32+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase. The composition may comprise (a) a CD16- and CD64+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase, (b) a CD16- and CD89+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase; and (c) a CD16- and CD32+ granulocyte or precursor thereof (e.g., a granulocyte precursor cell) which expresses neutrophil elastase.

[0136] The present disclosure includes definitions of granulocytes or precursors thereof with reference to a recited expression (either positive or negative) of a number of specified markers. As will be apparent, when referring to granulocytes or precursors thereof, the granulocytes or precursors thereof referenced may comprise different types of granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Accordingly, such definitions may be taken as requiring that the granulocytes or precursors thereof in question comprises cells that are positive or negative (as required by the definition) for the recited markers. For example, in the case of granulocytes or precursors thereof defined as positive for expression of a first marker, negative for expression of a second marker, and positive for expression of a third marker, this requirement may be met by granulocytes or precursors thereof that comprise cells positive for the first marker, while also comprising cells negative for the second marker, and further comprising cells positive for the third marker. In such an embodiment, the granulocytes or precursors thereof may be heterogeneous in respect of cells that have the recited expression (whether positive or negative). Cells that each exhibit the required expression in respect of each of the recited markers may make up the largest group of cells within such granulocytes or precursors thereof. Cells that each exhibit the required expression in respect of each of the recited markers may make up the majority of cells within such granulocytes or precursors thereof. Cells that each exhibit the required expression in respect of each of the recited markers may provide at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of cells within such granulocytes or precursors thereof.

[0137] Preferably, when referring to granulocytes or precursors thereof which are negative for a recited marker, all of the granulocytes or precursors thereof are negative forthe recited marker. Thus, in the case of granulocytes or precursors thereof defined as positive for expression of a first marker, negative for expression of a second marker, and positive for expression of a third marker, this requirement may preferably be met by granulocytes or precursors thereof that comprise cells positive for the first marker, and comprising cells positive for the third marker, wherein all of the granulocytes or precursors thereof which express the first or third markers are negative for the second marker. In one embodiment, a cell in the granulocytes or precursors thereof may express at least 2, 3, 4, or 5 of the recited markers. In one embodiment, each of the cells in the granulocytes or precursors thereof may express at least 2, 3, 4, or 5 of the recited markers.

[0138] Such definitions may be taken as requiring that the granulocytes or precursors thereof in question consist of cells that are positive or negative (as required by the definition) for the recited markers. In such an embodiment, the granulocytes or precursors thereof are homogeneous in respect of cells that have the recited expression (whether positive or negative).

[0139] The composition may comprise cells comprising granulocytes, or precursors thereof (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR), wherein:

[0140] • at least 50% of the cells express CD11 b,

[0141] • at least 50% of the cells express CD15,

[0142] • at least 50% of the cells express CD64,

[0143] • at least 50% of the cells express CD89,

[0144] • at least 50% of the cells express CXCR2,

[0145] • at least 50% of the cells express neutrophil elastase,

[0146] • less than 50% of the cells express CD14,

[0147] • less than 50% of the cells express CD19,

[0148] • less than 50% of the cells express CD3,

[0149] • less than 50% of the cells express CD34,

[0150] • less than 50% of the cells express CD66b,

[0151] • less than 50% of the cells express CD68,

[0152] • less than 50% of the cells express CXCR4, and / or

[0153] • less than 50% of the cells express HLA-DR.

[0154] The composition may comprise cells comprising granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein:

[0155] • at least 50% of the cells express CD11 b,

[0156] • at least 50% of the cells express CD15,

[0157] • at least 50% of the cells express CD64,

[0158] • at least 50% of the cells express CD89, • at least 50% of the cells express CXCR2,

[0159] • at least 50% of the cells express neutrophil elastase,

[0160] • less than 50% of the cells express CD14,

[0161] • less than 50% of the cells express CD19,

[0162] • less than 50% of the cells express CD3,

[0163] • less than 50% of the cells express CD34,

[0164] • less than 50% of the cells express CD66b,

[0165] • less than 50% of the cells express CD68,

[0166] • less than 50% of the cells express CXCR4, or

[0167] • less than 50% of the cells express HLA-DR.

[0168] Preferably, the composition may comprise granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein:

[0169] • at least 50% of the cells express CD11 b,

[0170] • at least 50% of the cells express CD15,

[0171] • at least 50% of the cells express CD64,

[0172] • at least 50% of the cells express CD89,

[0173] • at least 50% of the cells express CXCR2,

[0174] • at least 50% of the cells express neutrophil elastase,

[0175] • less than 50% of the cells express CD14,

[0176] • less than 50% of the cells express CD19,

[0177] • less than 50% of the cells express CD3,

[0178] • less than 50% of the cells express CD34,

[0179] • less than 50% of the cells express CD66b,

[0180] • less than 50% of the cells express CD68,

[0181] • less than 50% of the cells express CXCR4, and

[0182] • less than 50% of the cells express HLA-DR.

[0183] At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of the cells may express CD11 b. Approximately 50 to 100%, approximately 50 to 95%, approximately 50 to 90%, approximately 50 to 85%, approximately 50 to 80%, approximately 50 to 75%, or approximately 50 to 70% of the cells may express CD11 b. Approximately 50 to 65%, approximately 50 to 60%, approximately 50 to 55% of the cells may express CD11 b. Approximately 55 to 100%, approximately 60 to 100%, approximately 65 to 100%, approximately 70 to 100% of the cells may express CD11 b. Approximately 55 to 95%, approximately 55 to 100%, approximately 55 to 90%, approximately 55 to 85%, approximately 55 to 80%, approximately 55 to 75%, approximately 55 to 70%, approximately 55 to 65% of the cells may express CD11 b. Approximately 60 to 95%, approximately 60 to 90%, approximately 60 to 85%, approximately 60 to 80%, approximately 60 to 75%, approximately 60 to 70%, approximately 60 to 65% of the cells may express CD11 b. Approximately 65 to 95%, approximately 65 to 90%, approximately 65 to 85%, approximately 65 to 80%, approximately 65 to 75%, approximately 65 to 70% of the cells of the cells may express CD11 b. Preferably, 55 to 65% of the cells may express CD11 b.

[0184] At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the cells of the cells may express CD15. Approximately 50 to 100%, approximately 50 to 95%, approximately 50 to 90%, approximately 50 to 85%, approximately 50 to 80% of the cells may express CD15. Approximately 55 to 100%, approximately 60 to 100%, approximately 65 to 100%, approximately 70 to 100%, approximately 75 to 100%, approximately 80 to 100%, approximately 85 to 100% of the cells may express CD15. Approximately 70 to 95%, approximately 70 to 90%, approximately 70 to 85%, approximately 70 to 80%, approximately 70 to 75% of the cells of the cells may express CD15. Approximately 75 to 95%, approximately 75 to 90%, approximately 75 to 85%, approximately 75 to 80%, of the cells may express CD15. Approximately 80 to 95%, approximately 80 to 90%, approximately 80 to 85% of the cells may express CD15. Preferably, 70 to 90% of the cells may express CD15.

[0185] At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the cells may express CD64. Approximately 50 to 100%, approximately 50 to 95%, approximately 50 to 90%, approximately 50 to 85%, approximately 50 to 80% of the cells may express CD64. Approximately 55 to 100%, approximately 60 to 100%, approximately 65 to 100%, approximately 70 to 100%, approximately 75 to 100%, approximately 80 to 100%, approximately 85 to 100% of the cells may express CD64. Approximately 70 to 95%, approximately 70 to 90%, approximately 70 to 85%, approximately 70 to 80%, approximately 70 -75% of the cells of the cells may express CD64. Approximately 75 to 95%, approximately 75 to 90%, approximately 75 to 85%, approximately 75 to 80%, of the cells may express CD64. Approximately 80 to 95%, approximately 80 to 90%, approximately 80 to 85% of the cells may express CD64. Preferably, 70-90% of the cells may express CD64. At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the cells may express CD89. Approximately 50 to 100%, approximately 50 to 95%, approximately 50 to 90% of the cells of the cells may express CD89. Approximately 55 to 100%, approximately 60 to 100%, approximately 65 to 100%, approximately 70 to 100%, approximately 75 to 100%, approximately 80 to 100%, approximately 85 to 100%, approximately 90 to 100% of the cells may express CD89. Approximately 70 to 95%, approximately 75 to 95%, approximately 70 to 85%, approximately 75 to 85%, approximately 75 -80% of the cells may express CD89. Approximately 80 to 95%, approximately 85 to 95%, approximately 90 to 95%, approximately 80 to 90%, approximately 80 to 85%, approximately 85 to 90% of the cells may express CD89. Preferably, 75-95% of the cells may express CD89.

[0186] At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the cells may express CXCR2. Approximately 50 to 100%, approximately 50 to 95%, approximately 50 to 90%, approximately 50 to 85%, approximately 50 to 80%, approximately 50 to 75%, approximately 50 to 70%, approximately 50 to 65%, approximately 50 to 60%, approximately 50 to 55% of the cells may express CXCR2. Approximately 55 to 100%, approximately 60 to 100%, approximately 65 to 100%, approximately 70 to 100%, approximately 75 to 100% of the cells may express CXCR2. Approximately 55 to 95%, approximately 55 to 90%, approximately 55 to 85%, approximately 55 to 80%, approximately 55 to 75%, approximately 55 to 70%, approximately 55 to 65%, approximately 55 to 60% of the cells may express CXCR2. Approximately 60 to 95%, approximately 60 to 90%, approximately 60 to 85%, approximately 60 to 80%, approximately 60 to 75%, approximately 60 to 70%, approximately 60 to 65%, approximately 65 to 70% of the cells may express CXCR2. Preferably, 50-70% of the cells may express CXCR2.

[0187] At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% of the cells may express neutrophil elastase. Approximately 50 to 100%, approximately 50 to 95%, approximately 50 to 90%, approximately 50 to 85%, approximately 50 to 80%, approximately 50 to 75%, approximately 50 to 70%, approximately 50 to 65%, approximately 50 to 60% of the cells may express neutrophil elastase. Approximately 55 to 100%, approximately 60 to 100%, approximately 65 to 100%, approximately 70 to 100%, approximately 75 to 100%, approximately 80 to 100% of the cells may express neutrophil elastase. Approximately 55 to 95%, approximately 55 to 90%, approximately 55 to 85%, approximately 55 to 80%, approximately 55 to 75%, approximately 55 to 70%, approximately 55 to 65%, approximately 55 to 60% of the cells may express neutrophil elastase. Approximately 60 to 95%, approximately 65 to 95%, approximately 60 to 90%, approximately 65 to 90%, approximately 60 to 85%, approximately 65 to 85%, approximately 60 to 80%, approximately 65 to 80%, approximately 60 to 75%, approximately 65 to 70%, approximately 60 to 70%, approximately 60 to 65%, approximately 65 to 70%, approximately 70 to 80% of the cells may express neutrophil elastase. Preferably, 60 to 80% of the cells may express neutrophil elastase.

[0188] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD14. Approximately 0 to 2.5%, approximately 0 to 5%, approximately 0 to 7.5%, approximately 0 to 10%, approximately 0 to 12.5%, approximately 0 to 15% of the cells may express CD14. Approximately 2.5% - 10%, approximately 5% - 10%, approximately 7.5 to 10% of the cells may express CD14. Preferably, 0 to 10% of the cells may express CD14.

[0189] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD19. Approximately 0 to 1.25%, approximately 0 - 2.5%, approximately 0 to 5%, approximately 1.25 to 2.5% of the cells of the cells may express CD19. Preferably, 0 to 2.5% of the cells may express CD19.

[0190] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD3. Approximately 0 to 1.25%, approximately 0 to 2.5%, approximately 0 to 5%, approximately 1 .25 to 2.5% of the cells may express CD3. Preferably, 0 to 2.5% of the cells may express CD3.

[0191] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD34. Approximately 0 to 2.5%, approximately 0 to 5%, approximately 0 to 7.5%, approximately 0 to 10%, approximately 0 to 12.5%, approximately 0 to 15% of the cells may express CD34. Approximately 2.5% to 10%, approximately 5% to 10%, approximately 7.5 to 10% of the cells may express CD34. Preferably, 0 to 10% of the cells may express CD34.

[0192] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD56. Approximately 0 to 1.25%, approximately 0 to 2.5%, approximately 0 to 5%, approximately 1.25 to 2.5% of the cells of the cells may express CD56. Preferably, 0 to 2.5% of the cells may express CD56.

[0193] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD66b. Approximately 0 to 5%, approximately 0 to 10%, approximately 0 to 15%, approximately 0 to 20%, approximately 0 to 25%, approximately 0 to 30% of the cells may express CD66b. Approximately 5% to 10%, approximately 5 to 15%, approximately 5 to 20%, approximately 10 to 15%, approximately 15 to 20% of the cells may express CD66b. Preferably, 0 to 20% of the cells may express CD66b.

[0194] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CD68. Approximately 0 to 1.25%, approximately 0 to 2.5%, approximately 0 to 5%, approximately 1.25 to 2.5% of the cells may express CD68. Preferably, 0 to 2.5% of the cells may express CD68.

[0195] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express CXCR4. Approximately 15 to 50%, approximately 15 to 45%, approximately 15 to 40%, approximately 15 to 35%, approximately 15 to 30%, approximately 15 to 25%, approximately 15 to 20% of the cells may express CXCR4. Approximately 20 to 50%, approximately 20 to 45%, approximately 20 to 40%, approximately 20 to 35%, approximately 20 to 30%, approximately 20 to 25% of the cells may express CXCR4. Approximately 25 to 50%, approximately 25 to 45%, approximately 25 to 40%, approximately 25 to 35%, approximately 25 to 30% of the cells may express CXCR4. Preferably, 20 to 40% of the cells may express CXCR4.

[0196] Less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5% of the cells may express HLA-DR. Approximately 15 to 50%, approximately 15 to 45%, approximately 15 to 40%, approximately 15 to 35%, approximately 15 to 30%, approximately 15 to 25%, approximately 15 to 20% of the cells may express HLA-DR. Approximately 20 to 50%, approximately 20 to 45%, approximately 20 to 40%, approximately 20 to 35%, approximately 20 to 30%, approximately 20 to 25% of the cells may express HLA-DR. Approximately 25 to 50%, approximately 25 to 45%, approximately 25 to 40%, approximately 25 to 35%, approximately 25 to 30% of the cells may express HLA-DR. Preferably, 20 to 40% of the cells of the cells may express HLA-DR.

[0197] The composition may comprise cells comprising granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein:

[0198] • between 55% and 65% of the cells express CD11 b, and / or

[0199] • between 70% and 90% of the cells express CD15, and / or

[0200] • between 70% and 90% of the cells express CD64, and / or

[0201] • between 75% and 95% of the cells express CD89, and / or

[0202] • between 50% and 70% of the cells express CXCR2, and / or

[0203] • between 60% and 80% of the cells express neutrophil elastase, and / or

[0204] • between 0% and 10% of the cells express CD14, and / or

[0205] • between 0% and 2.5% of the cells express CD19, and / or

[0206] • between 0% and 2.5% of the cells express CD3, and / or

[0207] • between 0% and 10% of the cells express CD34, and / or

[0208] • between 0% and 20% of the cells express CD66b, and / or

[0209] • between 0% and 2.5% of the cells express CD68, and / or

[0210] • between 20% and 40% of the cells express CXCR4, and / or

[0211] • between 20% and 40% of the cells express HLA-DR.

[0212] Preferably, the composition may comprise cells comprising granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC- promoting FcR) wherein:

[0213] • between 55% and 65% of the cells express CD11 b,

[0214] • between 70% and 90% of the cells express CD15,

[0215] • between 70% and 90% of the cells express CD64,

[0216] • between 75% and 95% of the cells express CD89,

[0217] • between 50% and 70% of the cells express CXCR2,

[0218] • between 60% and 80% of the cells express neutrophil elastase,

[0219] • between 0% and 10% of the cells express CD14,

[0220] • between 0% and 2.5% of the cells express CD19,

[0221] • between 0% and 2.5% of the cells express CD3,

[0222] • between 0% and 10% of the cells express CD34, • between 0% and 20% of the cells express CD66b,

[0223] • between 0% and 2.5% of the cells express CD68,

[0224] • between 20% and 40% of the cells express CXCR4, and

[0225] • between 20% and 40% of the cells express HLA-DR.

[0226] The composition may comprise cells comprising granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein:

[0227] • at least 50% of the cells express CD64; and

[0228] • at least 50% of the cells express CD89; and / or neutrophil elastase.

[0229] The composition may comprise cells comprising granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein:

[0230] • at least 50% of the cells express CD64;

[0231] • at least 50% of the cells express CD89; and

[0232] • at least 50% of the cells express neutrophil elastase.

[0233] The composition may comprise cells comprising neutrophils, or precursors thereof, (e.g., neutrophils or precursors thereof which are CD16- and express an ADCC-promoting FcR) expressing neutrophil elastase, and wherein at least 50% of the cells express CD64; and / or CD89.

[0234] The composition may comprise cells comprising granulocytes, or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein more than 90% of the cells are Lin- (for example, approximately 97% of the cells are Lin-), and / or less than 30% of the cells are CD34+ cells (for example, approximately 14% of the cells are CD34+), and / or more than 10% of the cells are CD38+ (for example, approximately 12% of the cells are CD38+), and / or less than 1% of the cells have an HSC phenotype as defined above (for example approximately 0.04% of the cells have an HSC phenotype), and / or less than 1% of the cells have an LT-HSC phenotype as defined above (for example approximately 0.02% of the cells have an LT-HSC phenotype), and / or less than 20% of the cells have an LMPP phenotype as defined above (for example approximately 5% of the cells have an LMPP phenotype), and / or less than 10% of the cells have an MPP phenotype as defined above (for example approximately 2.5% of the cells have an MPP phenotype). The composition may comprise cells comprising granulocytes or precursors thereof, (e.g., granulocytes or precursors thereof which are CD16- and express an ADCC-promoting FcR) wherein more than 90% of the cells are Lin- cells (for example, approximately 97% of the cells are Lin- cells), and less than 30% of the cells are CD34+ (for example, approximately 14% of the cells are CD34+), and more than 10% of the cells are CD38+ (for example, approximately 12% of the cells are CD38+),and less than 1% of the cells have an HSC phenotype as defined above (for example approximately 0.04% of the cells have an HSC phenotype), and less than 1% of the cells have an LT-HSC phenotype as defined above (for example approximately 0.02% of the cells have an LT-HSC phenotype), and less than 20% of the cells have an LMPP phenotype as defined above (for example approximately 5% of the cells have an LMPP phenotype), and less than 10% of the cells have an MPP phenotype as defined above (for example approximately 2.5% of the cells have an MPP phenotype).

[0235] The composition may comprise a ratio of CD15- to CD15+ cells that is approximately 1 :1.

[0236] The composition may comprise around 25-75%, or 35-60 CD15- cells. For example, the composition may comprise approximately 50% CD15- cells.

[0237] The composition may comprise around 30-70%, or 40-65%, CD15+ cells. For example, the composition may comprise approximately 50% CD15+ cells.

[0238] The composition may comprise around 5-25%, 5-20%, 7-18%, or 10-15% CD15+CD66b+ cells. For example, the composition may comprise approximately 12% CD15+CD66b+ cells.

[0239] The composition may comprise around less than 30% or less than 25% CD11 b+ cells. For example, the composition may comprise approximately 10-25% or 15-25% CD11b+ cells, for example approximately 19% CD11 b+ cells.

[0240] The composition may comprise at least 30%, at least 35%, at least 40%, or at least 45% CD71 + cells. For example, the composition may comprise approximately 60% CD71+ cells.

[0241] The composition may comprise around 60-95%, or 65-90% CD49d+ cells. For example, the composition may comprise approximately 75% CD49d+ cells. The composition may comprise less than 5%, less than 4%, less than 3%, or less than 2% CD10+ cells. The composition may comprise around 0.03-2% CD10+ cells. For example, the composition may comprise approximately 0.5% CD10+ cells.

[0242] The composition may comprise around 1-12%, or 2-15% CD177+ cells. The composition may comprise approximately 6% CD177+ cells.

[0243] The composition may comprise less than 20% or less than 15% CD62L+ cells. For example, the composition may comprise between approximately 2-15%, for example approximately 8% CD62L+ cells.

[0244] The composition may comprise around 40-85%, or 50-75%, CD54+ cells. For example, the composition may comprise approximately 63% CD54+ cells.

[0245] The composition may comprise around 2-15%, or around 5-10% CD63+ cells. For example, the composition may comprise approximately 7% CD63+ cells.

[0246] The composition may comprise around 70-90%, or 75-85% CD18+ cells. For example, the composition may comprise approximately 80% CD18+ cells.

[0247] The composition may comprise around 35-55% HLA-DR+ cells. For example, the composition may comprise approximately 47% HLA-DR+ cells.

[0248] The composition may comprise around 6-8% CD115+ cells, For example, the composition may comprise approximately 5% CD115+ cells.

[0249] The composition may comprise around 5-30% CD40+ cells, For example, the composition may comprise approximately 15% CD40+ cells.

[0250] The composition may comprise around 20-55% CD32+ cells, For example, the composition may comprise approximately 40% CD32+ cells.

[0251] The composition may comprise around 4-9% CXCR2+ cells, For example, the composition may comprise approximately 6% CXCR2+ cells. The composition may comprise around 0.04-1% CD16+ cells. For example, the composition may comprise approximately 0.25% CD16+ cells.

[0252] The composition may comprise around 2-15% CD14+ cells. For example, the composition may comprise approximately 8% CD14+ cells.

[0253] The composition may comprise around 0.5-4% CD68+ cells. For example, the composition may comprise approximately 1.5% CD68+ cells.

[0254] The composition may comprise around 2-18% CD206+ cells. For example, the composition may comprise approximately 10% CD206+ cells.

[0255] Thus, the composition may comprise:

[0256] • more than 90% Lin- cells (for example, approximately 97% Lin- cells);

[0257] • less than 30% CD34+ cells (for example, approximately 14% CD34+ cells);

[0258] • more than 30% CD38+ cells (for example, approximately 65% CD38+ cells);

[0259] • less than 1% cells with an HSC phenotype (for example approximately 0.04% cells with an HSC phenotype);

[0260] • less than 1 % cells with an LT-HSC phenotype (for example approximately 0.02% cells with an LT-HSC phenotype;

[0261] • less than 20% cells with an LMPP phenotype (for example approximately 5% cells with an LMPP phenotype); and

[0262] • less than 10% cells with an MPP phenotype (for example approximately 2.5% cells with an MPP phenotype).

[0263] The composition may comprise granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+. The composition may comprise granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+. The granulocytes or precursors thereof which together are CD 15+ CD64+ CD 18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may also be positive for one, more than one, or all of the markers selected from the group comprising or consisting of: CD177, CD11 b, CD71 , CD66b, HLA-DR, CD115, CD49d, CD40, CD62L, CD54, CD18, CD34, CXCR4, CD64, CD32, CXCR2, CD38, Mac1 , 4-1 BBL, OX40L, PD-L1 , and CD14. The granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may express markers that closely resemble those expressed by neutrophil precursors. However, the granulocytes or precursors thereof may be CD 16- and CD62L-. This is in contrast to peripheral neutrophils found in the circulation and at times of homeostasis, which are CD16+ and CD62L+, thus providing a useful means by which the cells disclosed herein may be distinguished. The granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may be heterogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may be homogeneously positive for CD15, and heterogeneous in respect of the other markers of the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may be homogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein).

[0264] The granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may comprise granulocytes or precursors thereof which are 4- 1 BBL+ and / or OX40L+. These markers are ligands for T cells and NK cells, and their expression by these cells may indicate that the cells will have immunomodulatory activities. Similarly, the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may comprise granulocytes or precursors thereof which are CD38+ and / or CD40+ and / or CD54+, further co-stimulatory molecules associated with functional interactions with immune cells such as T cells. Accordingly, such cells may be effective in biological or therapeutic applications utilising the modulation of activity of such non- granulocytic inflammatory cell types.

[0265] In addition to expressing markers indicative of immunomodulatory ability, the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+) may comprise granulocytes or precursors thereof which express molecules (in particular CD11 b, CD18, Mac1 and CD32) that suggest they possess direct cytocidal activity. This may make these granulocytes or precursors thereof suitable for uses in which it is desired to therapeutically kill cells, such as cancerous or infected cells.

[0266] The composition may comprise granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR-. The composition may comprise granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-. These granulocytes or precursors thereof may comprise granulocytes or precursors thereof which are positive for one, more than one, or all of the markers selected from the group comprising or consisting of: CD177, CD11 b, CD71 , CD66b, CD115, CD49d, CD40, CD62L, CD54, CD18, CD34, CXCR4, CD64, CD32, CXCR2, CD38, Mac1 , 4-1 BBL, OX40L, PD-L1 , and CD14. The granulocytes or precursors thereof which together are CD15- CD11 b+Z- CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-) of cells may be heterogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-) of cells may be homogeneously negative for CD15 and HLA-DR, and heterogeneous in respect of the other markers of the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-) of cells may be homogeneously negative for CD15, HLA-DR and CD11 b, and heterogeneous in respect of the other markers of the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR- ) of cells may be homogenously positive for CD11 b and homogeneously negative for CD15 and HLA-DR, and heterogeneous in respect of the other markers of the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA- DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-) of cells may be homogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+Z- CD18+ CD49d+ CD32+ and HLA-DR-) may comprise granulocytes or precursors thereof which express markers, such as Mac-1 (comprising CD11 b and CD18) and CD32, that are consistent with a high capacity for cytotoxic activity. Accordingly, these granulocytes or precursors thereof may also be of benefit in medical uses or methods of treatment where direct cytocidal activity is required, such as the killing of cancerous or infected cells. The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-) may also comprise granulocytes or precursors thereof which express molecules such as 4-1 BBL and / or OX40L indicating their potential for immunomodulation, and suitability for use in biological or therapeutic applications requiring such activity. The granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g. the granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-) may also comprise granulocytes or precursors thereof which express CXCR2, which may be elevated by exposure of cells to IL-3 during methods in accordance with the invention, a marker that may contribute to heightened chemotaxis (in response to agents such as IL-8) and targeting of these cells into the tumour microenvironment.

[0267] The composition may comprise granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+. The composition may comprise granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+. These granulocytes or precursors thereof may comprise granulocytes or precursors thereof which are positive for one, more than one, or all of the markers selected from the group comprising or consisting of: CD177, CD71 , CD66b, CD115, CD49d, CD40, CD62L, CD54, CD18, CD34, CXCR4, CD64, CD32, CXCR2, CD38, Mac1 , 4-1 BBL, OX40L, PD-L1 , and CD14. The granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may be heterogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11b- HLA-DR+ CD18+ CD49d+ and CD71+) may be homogeneously negative for CD15 and CD11 b and homogenously positive for HLA-DR, and heterogeneous in respect of the other markers of the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may be homogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein).

[0268] The granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may comprise granulocytes or precursors thereof which express markers indicative of a relatively low level of differentiation. In keeping with this, the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may comprise granulocytes or precursors thereof which are CD34+. The granulocytes or precursors thereof which together are CD15- CD11b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may comprise cells expressing markers, such as 4-1 BBL and / or OX40L and / or CD40 and / or CD54 that indicate their suitability for use in applications requiring immunomodulation of non- granulocytic immune cells. The granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may comprise cells that have the capacity to differentiate further, and to express markers such as CD11 b and CD15 that would confer cytolytic activity. Accordingly, the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+) may be employed in medical uses or methods of treatment where in vivo signals would induce such differentiation, leading to the ability to kill deleterious cell types.

[0269] The composition may comprise granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA-DR+. The composition may comprise granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+. The granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA-DR+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+) may comprise cells that are positive for one, more than one, or all of the markers selected from the group comprising or consisting of: CD177, CD71 , CD66b, CD115, CD49d, CD40, CD62L, CD54, CD18, CD34, CXCR4, CD64, CD32, CXCR2, CD38, Mac1 , 4-1 BBL, OX40L, PD-L1 , and CD14. The granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA-DR+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+) may be heterogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA-DR+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+) may be homogeneously negative for CD15 and homogenously positive for HLA-DR and CD11 b, and heterogeneous in respect of the other markers of the recited marker profile (which may suitably include the optional constituents referred to herein). The granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA-DR+ (e.g., the granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+) may be homogeneous for the recited marker profile (which may suitably include the optional constituents referred to herein).

[0270] The composition may comprise:

[0271] • granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+);

[0272] • granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g., granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-);

[0273] • granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+); and / or

[0274] • granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA- DR+ (e.g., granulocytes or precursors thereof which together are CD15- CD11b+ and HLA-DR+).

[0275] The composition may comprise:

[0276] • granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+);

[0277] • granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g., granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-); • granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+); and optionally

[0278] • granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA- DR+ (e.g., granulocytes or precursors thereof which together are CD15- CD11b+ and HLA-DR+).

[0279] The composition may comprise:

[0280] • granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and / or CD71+ (e.g., granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+); and

[0281] • granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and / or HLA-DR- (e.g., granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-); and

[0282] • granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and / or CD71+ (e.g., granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+); and optionally

[0283] • granulocytes or precursors thereof which together are CD15- CD11 b+ and / or HLA- DR+ (e.g., granulocytes or precursors thereof which together are CD15- CD11b+ and HLA-DR+).

[0284] The composition may comprise:

[0285] • granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+;

[0286] • granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-;

[0287] • granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+; and / or

[0288] • granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+.

[0289] The composition may comprise:

[0290] • granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+; and

[0291] • granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-; and granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+

[0292] CD18+ CD49d+ and CD71+; and optionally granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+.

[0293] The composition may comprise:

[0294] • granulocytes or precursors thereof which together are CD15+ CD64+ CD18+ CD49d+ and CD71+; and

[0295] • granulocytes or precursors thereof which together are CD15- CD11 b+ / - CD18+ CD49d+ CD32+ and HLA-DR-; and

[0296] • granulocytes or precursors thereof which together are CD15- CD11 b- HLA-DR+ CD18+ CD49d+ and CD71+; and

[0297] • granulocytes or precursors thereof which together are CD15- CD11 b+ and HLA-DR+.

[0298] The composition may comprise granulocytes or precursors thereof which together are CD11 bhiCD15+ CD66b+ CD177+ CD18hiCD16- CD34- CD38- and / or CD49d-. The composition may comprise granulocytes or precursors thereof which together are CD11 bhiCD15+ CD66b+ CD177+ CD18hiCD16- CD34- CD38- and CD49d-.

[0299] The composition may comprise granulocytes or precursors thereof which together are CD34+Z- , CD38+ / -, CD15+ / -, CD49d+, CD18+, CD66b-, CD177-, and / or CD16-. The composition may comprise granulocytes or precursors thereof which together are CD34+ / -, CD38+ / -, CD15+ / -, CD49d+, CD18+, CD66b-, CD177-, and CD16-.

[0300] The composition may comprise a granulocyte (e.g., a terminally differentiated granulocyte, such as a neutrophil).

[0301] The granulocytes or precursors thereof described herein may be from any suitable source. The granulocytes or precursors thereof described herein may be obtained or obtainable from any suitable source. As used herein, reference to “obtained” may mean “obtainable”. Similarly, reference to “obtainable” may mean “obtained”.

[0302] Any of the cells disclosed herein may be derived from a mammal, such as a human, nonhuman primate, mouse, rat, dog, cat, horse, or cow. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be derived from a mammal, such as a human, nonhuman primate, mouse, rat, dog, cat, horse, or cow. The granulocyte or precursorthereof (e.g., a granulocyte precursor cell) may be derived from a human. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a human cell. Preferably, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be allogeneic with reference to the subject to be treated. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be autologous with reference to the subject to be treated.

[0303] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be obtainable (e.g. obtained) from a sample of PBMCs or a sample of umbilical cord blood. The sample of PBMCs or sample of umbilical cord blood may be obtainable (e.g. obtained) from a donor. The donor may be selected based on one or more of the following characteristics: sex, age, medical history, and / or blood group type. A donor may be selected if said donor is a healthy donor. A donor may be selected if said donor does not have cancer and does not have an infection. For example, a donor may be selected if said donor does not have cancer. A donor may be selected if said donor does not have an infection. A donor may be selected if said donor is a male. A donor may be selected if said donor is aged 18-55 and preferably 18-35 (more preferably 18-24). Suitably, a donor may be selected if said donor is a male aged between 18- 55 and preferably 18-35 (more preferably 18-24). In another embodiment a donor may be selected if said donor is a female. A donor may be selected if said donor is above the age of 40. Suitably, a donor may be selected if said donor is a female above the age of 40.

[0304] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be obtainable from (e.g. differentiated in vitro from) a cell, e.g., a stem cell, such as a haematopoietic stem cell or iPSC. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be isolated from a subject, or may be derived from a cell ex vivo. In particular, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be differentiated from a stem cell or a progenitor cell ex vivo. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be obtainable from a method of obtaining a granulocyte or precursor described herein.

[0305] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be differentiated from a stem cell or a progenitor cell using any suitable method. Preferably, the method may comprise culturing progenitor cells in cell culture conditions that promote differentiation of the progenitor cells to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Such a method may optionally comprise a further step of purifying the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) produced. Progenitor cells, and populations of progenitor cells, in the context of the present disclosure may usefully be defined by means of their expression of marker profiles and phenotypes. The following definitions, based upon suitable markers expression profiles, may be used singly or in combination to identify suitable populations of progenitor cells.

[0306] In a suitable embodiment, a population of progenitor cells comprises cells that are Lin- (as defined above). For example, a suitable population of progenitor cells may comprise at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% Lin- cells. By way of example, a suitable population of progenitor cells may comprise at least 98% Lin- cells. A suitable population of progenitor cells may comprise approximately 98-99% Lin- cells. Suitably, a population of progenitor cells comprises approximately 99% Lin- cells.

[0307] Alternatively, or additionally, a suitable population of progenitor cells comprises CD34+ cells. For example, such a population of progenitor cells may comprise between approximately 5- 90%, or approximately 10-85% CD34+ cells. By way of example, such a population of progenitor cells may comprise between approximately 15-80% CD34+ cells. In such an embodiment, the proportion of CD34+ cells may be between approximately 20-70%. Suitably, a population of progenitor cells comprises approximately 43% CD34+ cells.

[0308] Alternatively, or additionally, a suitable population of progenitor cells comprises CD38+ cells. For example, such a population of progenitor cells may between approximately 10-65%, approximately 15-60%, or approximately 20-55% CD38+ cells. By way of example, such a population of progenitor cells may comprise between approximately 25-50% CD38+ cells. In such an embodiment, the proportion of CD38+ cells may be between approximately 30% and 41%. Suitably, a population of progenitor cells comprises approximately 35% CD38+ cells.

[0309] Alternatively, or additionally, a suitable population of progenitor cells comprises cells with an HSC phenotype. For example, such a population of progenitor cells may comprise less than 5%, less than 4%, less than 3%, or less than 2% cells with an HSC phenotype. By way of example, such a population of progenitor cells may comprise less than 1% cells with an HSC phenotype. A suitable population of progenitor cells may comprise approximately 0.01-0.7% cells with an HSC phenotype. Suitably, a population of progenitor cells comprises approximately 0.3% cells with an HSC phenotype. Alternatively, or additionally, a suitable population of progenitor cells comprises cells with an LT-HSC phenotype. For example, such a population of progenitor cells may comprise less than 5%, less than 4%, less than 3%, or less than 2% cells with an LT-HSC phenotype. By way of example, such a population of progenitor cells may comprise less than 1% cells with an LT-HSC phenotype. A suitable population of progenitor cells may comprise approximately 0.01-0.03% cells with an LT-HSC phenotype. Suitably, a population of progenitor cells comprises approximately 0.02% cells with an LT-HSC phenotype.

[0310] Alternatively, or additionally, a suitable population of progenitor cells comprises cells with an LMPP phenotype. For example, such a population of progenitor cells may comprise less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, or less than 45% cells with an LMPP phenotype. By way of example, such a population of progenitor cells may comprise less than 40% cells with an LMPP phenotype. A suitable population of progenitor cells may comprise approximately 5-30% cells with an LMPP phenotype. Suitably, a population of progenitor cells comprises approximately 13% cells with an LMPP phenotype.

[0311] Alternatively, or additionally, a suitable population of progenitor cells comprises cells with an MPP phenotype. For example, such a population of progenitor cells may comprise less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, or less than 45% cells with an MPP phenotype. By way of example, such a population of progenitor cells may comprise less than 40% cells with an MPP phenotype. A suitable population of progenitor cells may comprise approximately 1-35% cells with an MPP phenotype. Suitably, a population of progenitor cells comprises approximately 13% cells with an MPP phenotype.

[0312] In a suitable embodiment, a population of progenitor cells may comprise more than 98% Lin- cells (for example, approximately 99% Lin- cells), and / or 15-18% CD34+ cells (for example, approximately 43% CD34+ cells), and / or 25-50% CD38+ cells (for example, approximately 35% CD38+ cells), and / or less than 1% cells with an HSC phenotype as defined above (for example approximately 0.3% cells with an HSC phenotype), and / or less than 1% cells with an LT-HSC phenotype as defined above (for example approximately 0.02% cells with an LT-HSC phenotype), and / or less than 40% cells with an LMPP phenotype as defined above (for example approximately 13% cells with an LMPP phenotype), and / or less than 40% cells with an MPP phenotype as defined above (for example approximately 13% cells with an MPP phenotype). In a suitable embodiment, a population of progenitor cells may comprise more than 98% Lin- cells (for example, approximately 99% Lin- cells), 15-18% CD34+ cells (for example, approximately 43% CD34+ cells), 25-50% CD38+ cells (for example, approximately 35% CD38+ cells), less than 1% cells with an HSC phenotype as defined above (for example approximately 0.3% cells with an HSC phenotype), less than 1% cells with an LT-HSC phenotype as defined above (for example approximately 0.02% cells with an LT-HSC phenotype), less than 40% cells with an LMPP phenotype as defined above (for example approximately 13% cells with an LMPP phenotype), and less than 40% cells with an MPP phenotype as defined above (for example approximately 13% cells with an MPP phenotype).

[0313] Alternatively, or additionally, a suitable population of progenitor cells may comprise a ratio of CD15- to CD15+ cells that is approximately 2:1 .

[0314] A suitable population of progenitor cells may comprise around 60-95% CD15- cells. For example, a suitable population of progenitor cells may comprise approximately 71% CD15- cells.

[0315] A suitable population of progenitor cells may comprise around 10-50% CD15+ cells. For example, a suitable population of progenitor cells may comprise approximately 35% CD15+ cells.

[0316] A suitable population of progenitor cells may comprise around 0.02-1% CD15+CD66b+ cells. For example, a suitable population of progenitor cells may comprise approximately 0.04-0.47% or 0.24% CD15+CD66b+ cells.

[0317] A suitable population of progenitor cells may comprise less than 20% CD11 b+ cells. For example, a suitable population of progenitor cells may comprise approximately 2-6%, or approximately 3% CD11 b+ cells.

[0318] A suitable population of progenitor cells may comprise around 25-60% CD71+ cells. For example, a suitable population of progenitor cells may comprise approximately 33% CD71 + cells.

[0319] A suitable population of progenitor cells may comprise around 90-100% CD49d+ cells. For example, a suitable population of progenitor cells may comprise approximately 95% CD49d+ cells. A suitable population of progenitor cells may comprise around 0.01-1.5% CD10+ cells. For example, a suitable population of progenitor cells may comprise approximately 0.5% CD10+ cells.

[0320] A suitable population of progenitor cells may comprise around 0.25-3% CD177+ cells. For example, a suitable population of progenitor cells may comprise approximately 1% CD177+ cells.

[0321] A suitable population of progenitor cells may comprise around 20-60%, or 40-60% CD62L+ cells. For example, a suitable population of progenitor cells may comprise approximately 46% CD62L+ cells.

[0322] A suitable population of progenitor cells may comprise around 1-17% CD54+ cells. For example, a suitable population of progenitor cells may comprise approximately 6% CD54+ cells.

[0323] A suitable population of progenitor cells may comprise around 2-20% CD63+ cells. For example, a suitable population of progenitor cells may comprise approximately 5% CD63+ cells.

[0324] A suitable population of progenitor cells may comprise around 70-90% CD18+ cells. For example, a suitable population of progenitor cells may comprise approximately 87% CD18+ cells.

[0325] The cell culture conditions that promote differentiation of the progenitor cells may comprise the presence of G-CSF, GM-CSF, IL-3 and TNF. The cell culture conditions that promote differentiation of the progenitor cells may further comprise the presence of at least one cytokine selected from the group consisting of: SCF, and TPO.

[0326] Cell culture conditions that promote differentiation of the progenitor cells may comprise Iscove’s modified Dulbecco’s medium (IMDM) as a cell culture medium. In a suitable embodiment, the IMDM may be a form of the medium that comprises high glucose, glutamine, HEPES, sodium pyruvate, and may optionally contain phenol red. The G-CSF may be provided at a concentration of 0.013 pg / mL, or more. For example, the G- CSF may be provided at a concentration of 0.016 pg / mL, or more, 0.02 pg / mL, or more, 0.03 pg / mL, or more, or 0.065 pg / mL, or more.

[0327] The G-CSF may be provided at a concentration of 0.65 pg / mL, or less. For example, the G- CSF may be provided at a concentration of 0.52 pg / mL, or less, 0.39 pg / mL, or less, or 0.26 pg / mL, or less.

[0328] The G-CSF may be provided at a concentration of approximately 0.013 pg / mL to 0.65 pg / mL, 0.016 pg / mL to 0.52 pg / mL, 0.02 pg / mL to 0.39 pg / mL, 0.03 pg / mL to 0.26 pg / mL, or 0.065 pg / mL to 0.195 pg / mL. The G-CSF may be provided at a concentration of approximately 0.13 pg / mL. The G-CSF may be provided at a concentration of 0.13 pg / mL.

[0329] Examples of suitable forms of G-CSF that may be used in this manner include the product produced by Peprotech, and the GMP product produced by BioLegend.

[0330] The GM-CSF may be provided at a concentration of 0.001 pg / mL, or more. For example, the GM-CSF may be provided at a concentration of 0.00125 pg / mL, or more, 0.00167 pg / mL, or more, 0.0025 pg / mL, or more, or 0.005 pg / mL, or more.

[0331] The GM-CSF may be provided at a concentration of 0.05 pg / mL, or less. For example, the GM-CSF may be provided at a concentration of 0.04 pg / mL, or less, 0.03 pg / mL, or less, or less, or 0.02 pg / mL, or less.

[0332] The GM-CSF may be provided at a concentration of approximately 0.001 pg / mL to 0.05 pg / mL, 00.125 pg / mL to 0.04 pg / mL, 0.00167 pg / mL to 0.03 pg / mL, 0.0025 pg / mL to 0.02 pg / mL, or 0.005 pg / mL to 0.015 pg / mL. In a suitable embodiment, the GM-CSF is provided at a concentration of approximately 0.01 pg / mL. Indeed, in a suitable embodiment, the GM-CSF is provided at a concentration of 0.01 pg / mL.

[0333] Examples of suitable forms of GM-CSF that may be used in this manner include the products produced by PeproTech and BioTechne, and the GMP product produced by BioTechne, details of which are set out in Table 2.

[0334] The GM-CSF may be provided to the cells for a period of between 24 and 72 hours, suitably a period of 48 hours during the cell culture conditions. For example, the GM-CSF may be provided to the cells for the final 48 hours of the period for which they are in culture. The GM- CSF may be provided to the cells on the second to fourth days of cell culture conditions that promote differentiation of the progenitor cells. The GM-CSF may be provided to the cells on the third to fifth days of cell culture conditions that promote differentiation of the progenitor cells.

[0335] The IL-3 may be provided at a concentration of 0.013 pg / mL, or more. For example, the IL-3 may be provided at a concentration of 0.016 pg / mL, or more, 0.02 pg / mL, or more, 0.03 pg / mL, or more, or 0.065 pg / mL, or more.

[0336] The IL-3 may be provided at a concentration of 0.65 pg / mL, or less. For example, the IL-3 may be provided at a concentration of 0.52 pg / mL, or less, 0.39 pg / mL, or less, or 0.26 pg / mL, or less.

[0337] The IL-3 may be provided at a concentration of approximately 0.013 pg / mL to 0.65 pg / mL, 0.016 pg / mL to 0.52 pg / mL, 0.02 pg / mL to 0.39 pg / mL, 0.03 pg / mL to 0.26 pg / mL, or 0.065 pg / mL to 0.195 pg / mL. The IL-3 may be provided at a concentration of approximately 0.13 pg / mL. The IL-3 may be provided at a concentration of 0.13 pg / mL.

[0338] Examples of suitable forms of IL-3 that may be used in this manner include the product produced by PeproTech, and the GMP product produced by PeproTech or BioTechne, details of which are set out in Table 2.

[0339] GM-CSF and IL-3 may be provided to the cells for a period of between 12 and 72 hours, suitably a period of 48 hours during the cell culture conditions. For example, GM-CSF and IL- 3 may be provided to the cells for the final 48 hours of the period for which they are in culture. GM-CSF and IL-3 may be provided to the cells on the fourth and fifth days of cell culture conditions that promote differentiation of the progenitor cells. GM-CSF and IL-3 may be provided to the cells on the third and fourth days of cell culture conditions that promote differentiation of the progenitor cells.

[0340] The methods of obtaining a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may make use of the cytokine tumour necrosis factor (TNF) as a supplement. The terms TNF and TNF-alpha are used interchangeably herein. The TNF may be provided at a concentration of 0.0001 pg / mL, or more. For example, the TNF may be provided at a concentration of 0.000125 pg / mL, or more, 0.000167 pg / mL, or more, 0.00025 pg / mL, or more, or 0.0005 pg / mL, or more.

[0341] The TNF may be provided at a concentration of 0.005 pg / mL, or less. For example, the TNF may be provided at a concentration of 0.004 pg / mL, or less, 0.003 pg / mL, or less, or 0.002 pg / mL, or less.

[0342] The TNF may be provided at a concentration of approximately 0.0001 pg / mL to 0.005 pg / mL, 0.000125 pg / mL to 0.004 pg / mL, 0.000167 pg / mL to 0.003 pg / mL, 0.00025 pg / mL to 0.002 pg / mL, or 0.0005 pg / mL to 0.0015 pg / mL. The TNF may be provided at a concentration of approximately 0.001 pg / mL. The TNF may be provided at a concentration of 0.001 pg / mL.

[0343] Examples of suitable forms of TNF that may be used in this manner include the product produced by PeproTech, and the GMP product produced by BioTechne, details of which are set out in Table 2.

[0344] The TNF may be provided to the cells for a period of between 12 and 36 hours, preferably a period of 24 hours during the cell culture conditions. For example, the TNF may be provided to the cells for the final 24 hours of the period for which they are in culture. The TNF may be provided to the cells on the third to fourth days of cell culture conditions that promote differentiation of the progenitor cells. The TNF may be provided to the cells on the fourth to fifth days of cell culture conditions that promote differentiation of the progenitor cells. The TNF may be provided to the cells on the fifth day of cell culture conditions that promote differentiation of the progenitor cells. The TNF may be provided to the cells on the fourth day of cell culture conditions that promote differentiation of the progenitor cells.

[0345] The method may optionally make use of the cytokine stem cell factor (SCF) as a supplement.

[0346] The SCF may be provided at a concentration of 0.013 pg / mL, or more. For example, the SCF may be provided at a concentration of 0.016 pg / mL, or more, 0.02 pg / mL, or more, 0.03 pg / mL, or more, or 0.065 pg / mL, or more.

[0347] The SCF may be provided at a concentration of 0.65 pg / mL, or less. For example, the SCF may be provided at a concentration of 0.52 pg / mL, or less, 0.39 pg / mL, or less, or 0.26 pg / mL, or less. The SCF may be provided at a concentration of approximately 0.013 pg / mL to 0.65 pg / mL, 0.016 pg / mL to 0.52 pg / mL, 0.02 pg / mL to 0.39 pg / mL, 0.03 pg / mL to 0.26 pg / mL, or 0.065 pg / mL to 0.195 pg / mL. The SCF may be provided at a concentration of approximately 0.13 pg / mL. The SCF may be provided at a concentration of 0.13 pg / mL.

[0348] Examples of suitable forms of SCF that may be used in this manner include the product produced by Peprotech, and the GMP product produced by PeproTech or BioTechne, details of which are set out in Table 2.

[0349] The method may optionally make use of the cytokine thrombopoietin (TPO) as a supplement.

[0350] The TPO may be provided at a concentration of 0.013 pg / mL, or more. For example, the TPO may be provided at a concentration of 0.016 pg / mL, or more, 0.02 pg / mL, or more, 0.03 pg / mL, or more, or 0.065 pg / mL, or more.

[0351] The TPO may be provided at a concentration of 0.65 pg / mL, or less. For example, the TPO may be provided at a concentration of 0.52 pg / mL, or less, 0.39 pg / mL, or less, or 0.26 pg / mL, or less.

[0352] The TPO may be provided at a concentration of approximately 0.013 pg / mL to 0.65 pg / mL, 0.016 pg / mL to 0.52 pg / mL, 0.02 pg / mL to 0.39 pg / mL, 0.03 pg / mL to 0.26 pg / mL, or 0.065 pg / mL to 0.195 pg / mL. The TPO may be provided at a concentration of approximately 0.13 pg / mL. The TPO may be provided at a concentration of 0.13 pg / mL.

[0353] Examples of suitable forms of TPO that may be used in this manner include the product produced by Peprotech, and the GMP products produced by BioTechne or Peprotech, details of which are set out in Table 2.

[0354] The cell culture conditions used in culturing the population of progenitor cells may further comprise the presence of at least one supplement selected from the group comprising or consisting of: insulin transferrin selenium (ITS), and human serum albumin (HSA). The cell culture conditions may comprise the presence of both ITS and HSA.

[0355] The method may make use of insulin at a concentration of between about 0.1 g / L and about 5g / L, for example at a concentration of approximately 1 .0 g / L, as a supplement. These methods may make use of transferrin at a concentration of between about 0.01 g / L and about 2.5g / L, for example at a concentration of approximately 0.55 g / L as a supplement. Such methods and cell culture media may make use of selenium at a concentration of between about 0.0001 g / L and about 0.003g / L, for example at a concentration of approximately 0.00067g / L, as a supplement.

[0356] The method may optionally make use of HSA as a supplement.

[0357] The HSA may be provided at a concentration of between 0.1% and 5%. For example, HSA may be provided as a supplement may be provided at a concentration of approximately 1%.

[0358] The cell culture conditions that promote differentiation of the progenitor cells may comprise: GM-CSF; and G-CSF; and SCF; and TPO; and IL-3; and TNF; and ITS; and HSA. The cell culture medium may comprise IMDM, optionally with Glutamax supplementation.

[0359] Thus, the cell culture conditions that promote differentiation of the progenitor cells may comprise: GM-CSF at a concentration of approximately 0.01 g / mL; and G-CSF at a concentration of approximately 0.13pg / mL; and SCF at a concentration of approximately 0.13pg / mL; and TPO at a concentration of approximately 0.13pg / mL; and IL-3 at a concentration of approximately 0.13pg / mL; and TNF at a concentration of approximately 0.001 pg / mL; and 1x ITS; and HSA at approximately 1%. The cell culture medium may comprise IMDM, optionally with Glutamax supplementation.

[0360] The method may comprise culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells for any suitable period of time. For example, the progenitor cells may be cultured for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days or 15 days in conditions to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The method may comprise culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells for a period of 1 to 7 days. For example, such methods may comprise culturing the cells in the relevant conditions for a period of 4 to 7 days. Such methods may comprise culturing the cells for approximately 1 day, or for approximately 2 days, or for approximately 3 days, or for approximately 4 days, or for approximately 5 days, or for approximately 6 days, or for approximately 7 days. The progenitor cells may be cultured for 1-10 days, 2-9 days, 3-8 days, 4-7 days, or 5-6 days in conditions to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The progenitor cells may be cultured for 4 days in conditions to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The progenitor cells may be cultured for 5 days in conditions to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The progenitor cells may be cultured for 6 days in conditions to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0361] The progenitor cells may be cultured at an initial seeding density of between approximately 1x105and 10x106cells per cm2.

[0362] The method may involve expansion of the number of cells present in the culture, such that the number of granulocytes or precursors thereof yielded by the method is larger than the number of progenitor cells present at the beginning of the method. The number of granulocytes or precursors thereof in the population produced may be increased, as compared to the number of progenitor cells present at the beginning of the method, by at least 1-fold, at least 2-fold, at least 3-fold, at least 4- fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, or at least 15-fold. The methods set out in the Examples produce granulocytes or precursors thereof that are approximately 3.5-fold larger in number than the initial population of progenitor cells.

[0363] The method may be practiced in respect of a population of progenitor cells that has been produced by in vitro expansion of a population of stem cells. Accordingly, such a method may further comprise a step of culturing a population of stem cells in cell culture conditions to produce the population of progenitor cells.

[0364] Accordingly, the method may further comprise a step of culturing a population of stem cells in cell culture conditions to produce the population of progenitor cells:

[0365] • wherein the cell culture conditions for producing the progenitor cells comprise the presence of

[0366] • SCF,

[0367] • Flt-3 Ligand,

[0368] • IL-3,

[0369] • IL-6, and

[0370] • TPO.

[0371] The number of progenitor cells produced in such a method may be markedly expanded as compared to the number of stem cells present at the start of the cell culture conditions. Merely by way of example, such an embodiment of a method of the invention may achieve an expansion of progenitor cell numbers that is at least 50-fold, at least 75-fold, at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, or at least 350-fold, or more, as compared to the number of stem cells at the start of the cell culture conditions. The Examples set out details of a protocol that the inventors have used to achieve an approximately 75-fold increase in progenitor cell numbers, as compared to the starting stem cell population.

[0372] Accordingly, the method of obtaining the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may comprise: a) culturing a population of stem cells in cell culture conditions for producing progenitor cells comprising the presence of:

[0373] • SCF,

[0374] • Flt-3 Ligand,

[0375] • IL-3,

[0376] • IL-6, and

[0377] • TPO; to produce a population of progenitor cells; and b) culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells comprising the presence of:

[0378] • G-CSF,

[0379] • GM-CSF,

[0380] • IL-3 and

[0381] • TNF; to produce the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ; and optionally c) harvesting the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0382] The total increase in number of cells achieved by such a method of the invention, representing the change in cell numbers from the initial population of stem cells to the granulocytes or precursors thereof produced, may be at least 50-fold, at least 100-fold, at least 150-fold, at least 200-fold, at least 250-fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 450-fold, at least 500-fold, at least 550-fold, at least 600-fold, at least 650-fold, at least 700- fold, at least 750-fold, at least 800-fold, at least 850-fold, at least 900-fold, at least 950-fold, at least 1000-fold, at least 1050-fold, at least 1100-fold, at least 1150-fold, at least 1200-fold, at least 1250-fold, or at least 1300-fold. The Examples set out details of a protocol that the inventors have used to achieve greater than 250-fold increase in granulocyte or precursor thereof (e.g., a granulocyte precursor cell) numbers, as compared to the starting stem cell population.

[0383] A method in accordance with such embodiments of the invention may involve a total period of time in culture of between 10 and 25 days, for example of between 11 and 20 days, such as 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, or 19 days.

[0384] SCF may optionally be provided as a supplement the method comprising a step of producing a population of progenitor cells.

[0385] The SCF may be provided at a concentration of 0.02 pg / mL, or more. For example, the SCF may be provided at a concentration of 0.025 pg / mL, or more, 0.03 pg / mL, or more, 0.05 pg / mL, or more, or 0.1 pg / mL, or more.

[0386] The SCF may be provided at a concentration of 1 pg / mL, or less. For example, the SCF may be provided at a concentration of 0.8 pg / mL, or less, 0.6 pg / mL, or less, or 0.4 pg / mL, or less.

[0387] The SCF may be provided at a concentration of approximately 0.02 pg / mL to 1 pg / mL, 0.025 pg / mL to 0.8 pg / mL, 0.03 pg / mL to 0.6 pg / mL, 0.05 pg / mL to 0.4 pg / mL, or 0.1 pg / mL to 0.3 pg / mL. The SCF may be provided at a concentration of approximately 0.2 pg / mL. The SCF may be provided at a concentration of 0.2 pg / mL.

[0388] The forms of SCF discussed above are also suitable for use in such embodiments.

[0389] Flt-3 ligand (F3L) may optionally be provided as a supplement in the method comprising a step of producing a population of progenitor cells.

[0390] The F3L may be provided at a concentration of 0.02 pg / mL, or more. For example, the F3L may be provided at a concentration of 0.025 pg / mL, or more, 0.03 pg / mL, or more, 0.05 pg / mL, or more, or 0.1 pg / mL, or more.

[0391] The F3L may be provided at a concentration of 1 pg / mL, or less. For example, the F3L may be provided at a concentration of 0.8 pg / mL, or less, 0.6 pg / mL, or less, or 0.4 pg / mL, or less.

[0392] The F3L may be provided at a concentration of approximately 0.02 pg / mL to 1 pg / mL, 0.025 pg / mL to 0.8 pg / mL, 0.03 pg / mL to 0.6 pg / mL, 0.05 pg / mL to 0.4 pg / mL, or 0.1 pg / mL to 0.3 pg / mL. The F3L may be provided at a concentration of approximately 0.2 pg / mL. The F3L may be provided at a concentration of 0.2 pg / mL.

[0393] Examples of suitable forms of F3L that may be used in this manner include the product produced by Peprotech, and the GMP product produced by PeproTech or BioTechne, details of which are set out in Table 2.

[0394] IL-3 may optionally be provided as a supplement in the method comprising a step of producing a population of progenitor cells.

[0395] The IL-3 may be provided at a concentration of 0.0015 pg / mL, or more. For example, the IL- 3 may be provided at a concentration of 0.0019 pg / mL, or more, 0.0025 pg / mL, or more, 0.00375 pg / mL, or more, or 0.0075 pg / mL, or more.

[0396] The IL-3 may be provided at a concentration of 0.075 pg / mL, or less. For example, the IL-3 may be provided at a concentration of 0.06 pg / mL, or less, 0.045 pg / mL, or less, or 0.03 pg / mL, or less.

[0397] The IL-3 may be provided at a concentration of approximately 0.0015 pg / mL to 0.075 pg / mL, 0.0019 pg / mL to 0.06 pg / mL, 0.0025 pg / mL to 0.045 pg / mL, 0.00375 pg / mL to 0.03 pg / mL, or 0.0075 pg / mL to 0.0225 pg / mL. The IL-3 may be provided at a concentration of approximately 0.015 pg / mL. The IL-3 may be provided at a concentration of 0.015 pg / mL.

[0398] The forms of IL-3 discussed above are suitable for use in such embodiments.

[0399] Interleukin 6 (IL-6) may optionally be provided as a supplement in the method comprising a step of producing a population of progenitor cells.

[0400] The IL-6 may be provided at a concentration of 0.0015 pg / mL, or more. For example, the IL- 6 may be provided at a concentration of 0.0019 pg / mL, or more, 0.0025 pg / mL, or more, 0.00375 pg / mL, or more, or 0.0075 pg / mL, or more.

[0401] The IL-6 may be provided at a concentration of 0.075 pg / mL, or less. For example, the IL-6 may be provided at a concentration of 0.06 pg / mL, or less, 0.045 pg / mL, or less, or 0.03 pg / mL, or less. The IL-6 may be provided at a concentration of approximately 0.0015 pg / mL to 0.075 pg / mL, 0.0019 pg / mL to 0.06 pg / mL, 0.0025 pg / mL to 0.045 pg / mL, 0.00375 pg / mL to 0.03 pg / mL, or 0.0075 pg / mL to 0.0225 pg / mL. The IL-6 may be provided at a concentration of approximately 0.015 pg / mL. The IL-6 may be provided at a concentration of 0.015 pg / mL.

[0402] Examples of suitable forms of IL-6 that may be used in this manner include the product produced by PeproTech, and the GMP product produced by PeproTech or BioTechne, details of which are set out in Table 2.

[0403] TPO may optionally be provided as a supplement in the method comprising a step of producing a population of progenitor cells.

[0404] The TPO may be provided at a concentration of 0.002 pg / mL, or more. For example, the TPO may be provided at a concentration of 0.0025 pg / mL, or more, 0.003 pg / mL, or more, 0.005 pg / mL, or more, or 0.01 pg / mL, or more.

[0405] The TPO may be provided at a concentration of 0.1 pg / mL, or less. For example, the TPO may be provided at a concentration of 0.08 pg / mL, or less, 0.06 pg / mL, or less, or 0.04 pg / mL, or less.

[0406] The TPO may be provided at a concentration of approximately 0.002 pg / mL to 0.1 pg / mL, 0.0025 pg / mL to 0.08 pg / mL, 0.003 pg / mL to 0.06 pg / mL, 0.005 pg / mL to 0.04 pg / mL, or 0.01 pg / mL to 0.03 pg / mL. The TPO may be provided at a concentration of approximately 0.02 pg / mL. The TPO may be provided at a concentration of 0.02 pg / mL.

[0407] The forms of TPO discussed above are also suitable for use in these embodiments.

[0408] The cell culture conditions that promote production of progenitor cells may comprise: SCF; and Flt-3 Ligand; and IL-3; and IL-6; and TPO; and ITS; and HSA.

[0409] Thus, the cell culture conditions that promote production of progenitor cells may comprise: SCF at a concentration of approximately 0.2pg / mL; and Flt-3 Ligand at a concentration of approximately 0.2pg / mL; and IL-3 at a concentration of approximately 0.015pg / mL; and IL-6 at a concentration of approximately 0.015pg / mL; and TPO at a concentration of approximately 0.02pg / mL; and 1x ITS; and HSA at approximately 1%. Stem cells that may be employed in such methods, as a starting material for the production of progenitor cells (and ultimately granulocytes or precursors thereof) include, but are not limited to, HSCs and induced pluripotent stem cells (iPSCs). Accordingly, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be differentiated ex vivo from an HSC or an iPSC. In particular, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be differentiated ex vivo from an HSC. Further sources of stem cells include cord blood and mobilized blood.

[0410] The cell culture conditions used in culturing the stem cells to produce progenitor cells may further comprise the presence of at least one supplement selected from the group comprising or consisting of: ITS, and HSA. Such cell culture conditions may comprise the presence of both ITS and HSA.

[0411] ITS may be provided as a supplement in the method comprising a step of producing a population of progenitor cells.

[0412] Such embodiments may make use of insulin at a concentration of between about 0.1 g / L and about 5g / L, for example at a concentration of approximately 1 .0 g / L, as a supplement. These methods may make use of transferrin at a concentration of between about 0.01 g / L and about 2.5g / L, for example at a concentration of approximately 0.55 g / L as a supplement. Such methods may make use of selenium at a concentration of between about 0.0001 g / L and about 0.003g / L, for example at a concentration of approximately 0.00067g / L, as a supplement.

[0413] HSA may be provided as a supplement in the methods comprising a step of producing a population of progenitor cells.

[0414] The HSA may be provided at a concentration of between 0.1% and 5%. For example, HSA provided as a supplement may be provided at a concentration of approximately 1%.

[0415] In the methods in which stem cells are cultured to yield progenitor cells, this may involve expansion of the number of cells present in the culture.

[0416] The method may comprise culturing a population of stem cells in cell culture conditions to produce a population of progenitor cells for any suitable period of time. For example, the cells may be cultured for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days or 15 days in conditions to produce a population of progenitor cells. Preferably the cells may be cultured for 8 or 9 days in conditions to produce a population of progenitor cells. The stem cells may be cultured for 1-15 days, 1-10 days, 2- 14 days, 3-13 days, 4-12 days, 5-11 days, 6-10 days, 7-9 days or 8-9 days in conditions to produce a population of progenitor cells. Preferably, the stem cells, such as HSCs, may be cultured for 8-9 days in conditions to produce a population of progenitor cells.

[0417] The stem cells may be cultured in conditions to produce the population of progenitor cells for a period of 6 to 10 days. For example, such methods may comprise culturing the cells for a period of 7 to 8 days. Such methods may comprise culturing the cells in cell culture conditions to produce a population of progenitor cells for approximately 6 days, or for approximately 7 days, or for approximately 8 days, or for approximately 9 days, or for approximately 10 days.

[0418] Accordingly, the method may comprise:

[0419] (a) culturing a population of stem cells in cell culture conditions for producing progenitor cells comprising the presence of SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA for 6-10 days, or preferably 8 days, to produce a population of progenitor cells; and

[0420] (b) culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells to obtain the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0421] The method may comprise:

[0422] (a) culturing a population of stem cells in cell culture conditions for producing progenitor cells comprising the presence of IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA for 6-10 days, or preferably 8 days, to produce a population of progenitor cells; and

[0423] (b) culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells comprising IMDM, G-CSF, GM-CSF, IL-3, and TNF for 1- 6 days, or preferably 5 days, to obtain the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0424] The method may comprise:

[0425] (a) culturing a population of stem cells in cell culture conditions for producing progenitor cells comprising the presence of IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA for 6-10 days, or preferably 8 days, to produce a population of progenitor cells; and

[0426] (b) culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells to obtain the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . For example, the method may comprise:

[0427] (a) culturing a population of stem cells in cell culture conditions for producing progenitor cells comprising the presence of IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA for 6-10 days, or preferably 8 days, to produce a population of progenitor cells; and

[0428] (b) culturing the population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells comprising IMDM, SCF, TPO, GCSF, ITS and HSA for 1- 6 days, or preferably 5 days, to obtain the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0429] Appropriately supplemented cell culture medium may be replaced or replenished at any suitable time during the culture of the stem cells in conditions for producing progenitor cells. For example, the cell culture medium may be replenished on day 1 , day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11 , day 12, day 13, day 14, or day 15 of culture of the stem cells. The cell culture medium may be replenished on day 1 and day 6 of culture of the stem cells. The cell culture medium may be replaced on day 1 , day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11 , day 12, day 13, day 14, or day 15 of culture of the stem cells. Preferably, the cell culture medium may be replaced on day 4 of culture of the stem cells.

[0430] Stem cells, such as HSCs, from which progenitor cells are to be produced may be seeded at any suitable cell density. For example, the stem cells may be seeded at a density of 1x105cells / mL- 1x106cells / mL, 2.5x105cells / mL - 1x106cells / mL, 3x105cells / mL - 8x105cells / mL or 4x105cells / mL - 6x105cells / mL, preferably 5x105cells / mL. The stem cells may be seeded at a density of 1x105cells / cm2- 1x106cells / cm2, 2.5x105cells / cm2- 1x106cells / cm2, 3x105cells / cm2- 8x105cells / cm2or 4x105cells / cm2- 6x105cells / cm2, preferably 5x105cells / cm2. Preferably, the stem cells (such as HSCs) may be seeded at a density of 5x105cells / mL and 5x105cells / cm2.

[0431] The cells may be seeded in any suitable culture vessel. For example, the cells may be seeded in a G-Rex 6M or G-Rex 10M culture vessel. The cells may be transferred to a new culture vessel at any suitable time. The cells may be sequentially transferred into cell culture vessels of increasing surface area. Such transfers may take place on day 1 , day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11 , day 12, day 13, day 14 or day 15 of the culture to produce progenitor cells. For example, the stem cells (such as HSCs) may be transferred from a smaller G-Rex to a G-Rex 10OM on day 1 , day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9, day 10, day 11 , day 12, day 13, day 14 or day 15 of the culture to produce progenitor cells. For example, the stem cells (such as HSCs) may be transferred to a G-Rex 100M, or a larger cell culture vessel such as a G-Rex 500M, on day 4 of expansion. Progenitor cells may be transferred to a new culture vessel on day 1 , day 2, day 3, day 4, day 5, day 6, day 7, day 8, day 9 or day 10 of the culture conditions that promote differentiation of progenitor cells to granulocytes or precursors thereof.

[0432] The method may comprise:

[0433] (a) seeding stem cells (such as HSCs) at 5x105cells / mL and 5x105cells / cm2;

[0434] (b) culturing the cells in cell culture medium comprising IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA for 8 days to obtain a population of progenitor cells, wherein the cell culture medium comprising IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA is replenished on day 1 and day 6 of such culture, and wherein the cell culture medium comprising IMDM, SCF, FLT- 3, TPO, IL-3, IL-6, ITS and HSA is replaced on day 4 of such culture;

[0435] (c) culturing the population of progenitor cells in a cell culture medium comprising IMDM, SCF, TPO, GCSF, ITS and HSA for 5-6 days to obtain the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , wherein the cell culture medium comprising IMDM, SCF, TPO, GCSF, ITS and HSA is replenished on day 3 of differentiation.

[0436] The method may comprise:

[0437] (a) seeding stem cells (such as HSCs) at 5x105cells / mL and 5x105cells / cm2;

[0438] (b) culturing the cells in cell culture medium comprising IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA for 8 days to obtain a population of progenitor cells, wherein the cell culture medium comprising IMDM, SCF, FLT-3, TPO, IL-3, IL-6, ITS and HSA is replenished on day 1 and day 6 of such culture, and wherein the cell culture medium comprising IMDM, SCF, FLT- 3, TPO, IL-3, IL-6, ITS and HSA is replaced on day 4 of such culture;

[0439] (c) culturing the population of progenitor cells in a cell culture medium comprising IMDM, SCF, TPO, G-CSF, ITS and HSA for 5-6 days to obtain the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , wherein the cell culture medium comprising IMDM, SCF, TPO, GCSF, ITS, HAS, GM-CSF, IL-3 and TNF is replenished on day 3 of differentiation.

[0440] The method of obtaining the granulocyte or precursor may comprise a step of culturing a population of stem cells in cell culture conditions to produce the population of progenitor cells, wherein the cell culture conditions for producing the progenitor cells comprise the presence of

[0441] SCF;

[0442] FLT-3 ligand; TPO; and a pyrimido-[4,5-b]-indole derivative that promotes the expansion of hematopoietic stem cells.

[0443] The number of cells may increase during this period of cell culture. Accordingly, the period of culturing a population of stem cells in cell culture conditions to produce a population of progenitor cells may be referred to as an “expansion phase” or “expansion step”. An expansion phase as described herein may also be used in other methods in accordance with the present invention, such as methods of the third aspect of the invention

[0444] The number of progenitor cells may expand by at least 1-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 30-fold, at least 35-fold, at least 40-fold, or at least 45-fold as compared to the number of stem cells present at the start of the cell culture. Indeed, the number of progenitor cells may expand by at least 50-fold, at least 100-fold, at least 150-fold, at least 200-fold, at least 250- fold, at least 300-fold, at least 350-fold, at least 400-fold, at least 450-fold, at least 500-fold, at least 550-fold, at least 600-fold, at least 650-fold, at least 700-fold, at least 750-fold, at least 800-fold, at least 850-fold, at least 900-fold, at least 950-fold, at least 1000-fold as compared to the number of stem cells present at the start of the cell culture.

[0445] The basal medium employed in an expansion step in accordance with the invention may be Iscove’s modified Dulbecco’s medium (IMDM).

[0446] The pyrimido-[4,5-b]-indole derivative may be provided at a concentration within the range of 5nM - 1.5pM. For example, the pyrimido-[4,5-b]-indole derivative may be provided at a concentration within the range of 15nM - 1.5pM.

[0447] The cell culture conditions for producing the progenitor cells may further comprise:

[0448] • ITS; and / or

[0449] • HSA.

[0450] The cell culture conditions for producing the progenitor cells may comprise one or more conditions independently selected from the group consisting of:

[0451] • SCF at a concentration of approximately 0.1 -0.2 pg / ml;

[0452] • FLT-3 ligand at a concentration of approximately 0.1-0.2 pg / ml;

[0453] • TPO at a concentration of approximately 0.01-0.02 pg / ml; and HSA at a concentration of approximately 1%.

[0454] Suitably the cell culture conditions for producing the progenitor cells may further comprise:

[0455] • IL-3; and / or

[0456] • IL-6.

[0457] By way of example, the cell culture conditions for producing the progenitor cells may comprise:

[0458] • IL-3 at a concentration of approximately 0.0001-0.015 pg / ml; and / or

[0459] • IL-6 at a concentration of approximately 0.0001-0.015 pg / ml.

[0460] The pyrimido-[4,5-b]-indole derivative used in the cell culture conditions for producing the progenitor cells may be selected from the group consisting of: UM 171 , UM729.

[0461] The cell culture conditions for producing the progenitor cells may comprise the pyrimido-[4,5- b]-indole derivative UM171 at a concentration within the range of 15nM - 250nM. The cell culture conditions may comprise UM 171 at a concentration within the range of 25nM - 100nM.

[0462] The cell culture conditions for producing the progenitor cells may comprise the pyrimido-[4,5- b]-indole derivative UM729 at a concentration within the range of 250nM - 1.5pM. For example, the cell culture conditions may comprise UM729 at a concentration within the range of 500nM - 1 pM.

[0463] Incidences on which a pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to cells may be separated by up to 5 days, up to 4 days, up to 3 days, up to 2 days, or by a single day. For example, different incidences of providing a pyrimido-[4,5-b]-indole derivative (such as UM729 or UM 171) may be separated by approximately 120 hours, by approximately 96 hours, by approximately 72 hours, by approximately 48 hours, or by approximately 24 hours.

[0464] The pyrimido-[4,5-b]-indole derivative may be provided to the cells in multiple incidences during the culture to produce the progenitor cells. For example, the pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to the cells in 1 - 10 incidences during the culture to produce the progenitor cells.

[0465] The pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to the cells in 4 incidences during the expansion phase. Suitably, the pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to the cells in 3 incidences during the expansion phase.

[0466] The pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to the cells in a first incidence 24 hours post-seeding.

[0467] The pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to the cells in a second incidence 72 hours after the first incidence.

[0468] The pyrimido-[4,5-b]-indole derivative (such as UM729 or UM171) may be provided to the cells in a third incidence 48 hours after the second incidence.

[0469] The pyrimido-[4,5-b]-indole derivative may be provided to the cells at the first incidence one day post-seeding.

[0470] The pyrimido-[4,5-b]-indole derivative may be provided to the cells at the second incidence four days post-seeding.

[0471] The pyrimido-[4,5-b]-indole derivative may be provided to the cells at the third incidence six days post-seeding.

[0472] The pyrimido-[4,5-b]-indole derivative may be provided in the first incidence at a concentration within the range of 15nM - 1.5pM.

[0473] In such an embodiment employing UM 171 , UM 171 may be provided in the first incidence at a concentration within the range of 25 - 100nM. For example, UM171 may be provided in the first incidence at a concentration of approximately 100nM.

[0474] In such an embodiment employing UM729, UM729 may be provided in the first incidence at a concentration within the range of 500 nM - 1 pM. For example, UM729 may be provided in the first incidence at a concentration of approximately 500 nM.

[0475] In a suitable embodiment the pyrimido-[4,5-b]-indole derivative may be provided in a second incidence at a concentration within the range of 15nM - 1 ,5pM. In such an embodiment employing UM 171 , UM 171 may be provided in a second incidence at a concentration within the range of 25 - 100 nM. For example, UM171 may be provided in the second incidence at a concentration of approximately 50 nM.

[0476] In such an embodiment employing UM729, UM729 may be provided in the second incidence at a concentration within the range of 500 nM - 1 pM. For example, UM729 may be provided in the second incidence at a concentration of approximately 500 nM.

[0477] In a suitable embodiment, the pyrimido-[4,5-b]-indole derivative may be provided in a third incidence at a concentration within the range of 15nM - 1 ,5pM.

[0478] In such an embodiment employing UM 171 , UM 171 may be provided in the third incidence at a concentration within the range of 25 - 100 nM. For example, UM171 may be provided in the third incidence at a concentration of approximately 100 nM.

[0479] In such an embodiment employing UM729, UM729 may be provided in the third incidence at a concentration within the range of 500 nM - 1 pM. For example, UM729 may be provided in the third incidence at a concentration of approximately 1 pM.

[0480] Suitably, the concentration of the pyrimido-[4,5-b]-indole derivative (such as UM729 or UM 171) used in the cell culture conditions for producing the progenitor cells may be the same on each incidence of administration.

[0481] When present, the concentration of the pyrimido-[4,5-b]-indole derivative (such as UM729 or UM 171) may be kept substantially constant throughout the cell culture conditions for producing the progenitor cells.

[0482] The method may further comprise a step of culturing the progenitor cells to promote their differentiation into granulocytes, or granulocyte precursor cells.

[0483] The method of obtaining the granulocyte or precursor may comprise culturing a population of progenitor cells in cell culture conditions that promote differentiation of the progenitor cells into granulocytes, or precursors thereof, the cell culture conditions comprising the presence of: a basal medium;

[0484] SCF;

[0485] TPO; and G-CSF.

[0486] The cell culture conditions which bring about the differentiation of progenitor cells to granulocytes, or precursors thereof, used in these methods may be referred to as a “differentiation phase” or “differentiation step”.

[0487] In a suitable embodiment, the population of progenitor cells may have been produced by a method comprising a step of culturing a population of stem cells in cell culture conditions to produce the population of progenitor cells, wherein the cell culture conditions for producing the progenitor cells comprise the presence of

[0488] • SCF;

[0489] • FLT-3 ligand;

[0490] • TPO; and

[0491] • a pyrimido-[4,5-b]-indole derivative that promotes the expansion of hematopoietic stem cells.

[0492] In the population of progenitor cells:

[0493] • at least 50% of the cells of the population may express CD34;

[0494] • at least 50% of the cells of the population may express CD13;

[0495] • at least 50% of the cells of the population may express HLA-DR;

[0496] • at least 50% of the cells of the population may express CD49d;

[0497] • at least 50% of the cells of the population may express CD33;

[0498] • less than 50% of the cells of the population may express CD15;

[0499] • less than 50% of the cells of the population may express CD38; and

[0500] • less than 50% of the cells of the population may express CD45RA.

[0501] By way of example, in the population of progenitor cells:

[0502] • between 80% and 90% of the cells of the population may express CD34; and / or

[0503] • between 90% and 100% of the cells of the population may express CD13; and / or

[0504] • between 70% and 85% of the cells of the population may express HLA-DR; and / or

[0505] • between 95% and 100% of the cells of the population may express CD49d; and / or

[0506] • between 95% and 10% of the cells of the population may express CD33; and / or

[0507] • between 0% and 10% of the cells of the population may express CD15; and / or

[0508] • between 20% and 50% of the cells of the population may express CD38; and / or

[0509] • between 15% and 30% of the cells of the population may express CD45RA. In the population of progenitor cells:

[0510] • between 80% and 90% of the cells of the population may express CD34;

[0511] • between 90% and 100% of the cells of the population may express CD13;

[0512] • between 70% and 85% of the cells of the population may express HLA-DR;

[0513] • between 95% and 100% of the cells of the population may express CD49d;

[0514] • between 95% and 10% of the cells of the population may express CD33;

[0515] • between 0% and 10% of the cells of the population may express CD15;

[0516] • between 20% and 50% of the cells of the population may express CD38; and

[0517] • between 15% and 30% of the cells of the population may express CD45RA.

[0518] Each of SCF, TPO, and G-CSF may be provided in first and second incidences of administration during the culturing in conditions that promote differentiation of progenitors into granulocytes, or precursors thereof. The amount of each of SCF, TPO, and G-CSF provided in the second incidence of administration may be approximately double that provided in the first incidence of administration. By way of example, the amount of each of SCF, TPO, and G- CSF provided in the first incidence of administration may be approximately 0.333 ng per 1000 cells. The amount of each of SCF, TPO, and G-CSF provided in the second incidence of administration may be approximately 0.666 ng per 1000 cells.

[0519] The first incidence of administration of SCF, TPO, and G-CSF may be on the first day of culturing in cell culture conditions that promote differentiation. The second incidence of administration of SCF, TPO, and G-CSF may be approximately 48 hours after the first incidence.

[0520] The cell culture conditions that promote differentiation of the progenitor cells into granulocytes, or precursors thereof, may further comprise at least one supplement from the group consisting of:

[0521] • GM-CSF

[0522] • IL-3

[0523] • TNFa

[0524] The cell culture conditions that promote differentiation of the progenitor cells into granulocytes, or precursors thereof, may comprise one, two or three of the following:

[0525] • GM-CSF at a concentration of approximately 0.01 pg / ml;

[0526] • IL-3 at a concentration of approximately 0.13 pg / ml; and TNFa at a concentration of approximately 0.001 pg / ml.

[0527] The cell culture conditions that promote differentiation of the progenitor cells into granulocytes, or precursors thereof, may comprise each of the following:

[0528] • GM-CSF at a concentration of approximately 0.01 pg / ml; and

[0529] • IL-3 at a concentration of approximately 0.13 pg / ml; and

[0530] • TNFa at a concentration of approximately 0.001 pg / ml.

[0531] GM-CSF and IL-3 may be provided to the cells for the final 48 hours of the period of culture in conditions that promote differentiation of the progenitor cells into granulocytes, or precursors thereof. In contrast, TNFa may be provided to the cells for the final 24 hours of the period of culture in conditions that promote differentiation of the progenitor cells into granulocytes, or precursors thereof.

[0532] The basal medium employed in a differentiation step may be Stemline II. In an alternative embodiment, the basal medium employed in a differentiation step in accordance with the invention may be IMDM. Alternatively, in one embodiment, the basal medium is not IMDM.

[0533] The differentiation step may achieve an at least 300-fold expansion cell numbers. Indeed, such a step may achieve an at least 450-fold expansion of cell numbers. Such expansions may be achieved in the differentiation phase, or in a combination of an expansion phase (if present) and the differentiation phase.

[0534] A “granulocyte progenitor cell” as used herein may be a cell more differentiated than an HSC but less differentiated than a granulocyte precursor cell.

[0535] The population of stem cells (e.g. HSCs) may be derived from any suitable source of stem cells (preferably a source of stem cells from human origin). Preferably, the population of stem cells (e.g. HSCs) is derived or obtained from a donor, preferably a human donor, having granulocytes (preferably neutrophils) with high cancer killing activity (CKA) (e.g. exhibiting high CKA, e.g. by for instance lysing cancer cells). Granulocytes (preferably neutrophils) may be isolated from peripheral blood of a donor or by leukapheresis using known techniques in the art. Any (in vitro) methods suitable to assess or measure CKA of granulocytes (preferably neutrophils) obtained from a (human) donor may be used. Preferably, the CKA of the granulocytes (preferably neutrophils) is assessed by a method comprising: b. admixing granulocytes (preferably neutrophils) from a donor with cancer cells; c. incubating said admixture; and d. measuring the % of cancer cells killed in said admixture, thereby assessing the CKA of said granulocytes (preferably neutrophils).

[0536] The % of cancer cells killed in the admixture in step (c) may be assessed by counting the number of cancer cells left in the admixture at the end of the incubation period (in step b) compared to the number of cancer cells present before admixing the granulocytes (preferably neutrophils) obtained from the (human) donor. For instance, if granulocytes (preferably neutrophils) are added to 100 cancer cells to form an admixture, and 25 cancer cells are left in said admixture at the term of the incubation period (in step b), then the percentage (%) of cancer cells killed is 75%.

[0537] Alternatively, the following formula may be used to determine the percentage (%) of cancer cells killed.

[0538] Percentage (%) of cells killed = cell number - Remaining cell number) x 100 Initial ceil number

[0539] A granulocyte (preferably a neutrophil) with high CKA may be a granulocyte (preferably a neutrophil) having a CKA of at least 30%, for instance at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or more.

[0540] Preferably, a granulocyte (preferably a neutrophil) with high CKA is a granulocyte (preferably a neutrophil) having a CKA of at least 30%, preferably at least 50%, more preferably at least 70%.

[0541] The present inventors have shown that the cancer killing efficacy (or cancer killing activity) of granulocytes (e.g. neutrophils) is genetically-defined, rather than epigenetically-defined. This is demonstrated in e.g., WO2019081879A1 , which shows that granulocytes (preferably neutrophils) derived (differentiated in vitro) from HSCs isolated (obtainable or obtained) from a (human) donor have similar cancer killing efficacy (cancer killing activity, e.g. ability to lyse cancer cells) to mature granulocytes isolated (e.g. freshly isolated from blood) directly from the same donor. Advantageously, donors found to have granulocytes (preferably neutrophils) with a high cancer killing activity can be used as a source of HSCs, which can be differentiated in vitro) into granulocyte precursors with similarly high cancer killing activity. Said granulocyte precursors will, in turn, differentiate into granulocytes (preferably neutrophils) in vitro as well as in vivo overtime following administration to an organism (e.g. mice) ora subject (e.g. human subject), and based on the same genetic mechanisms, will exhibit a similarly high cancer killing activity. This is beneficial as such HSCs can advantageously be stored and used for the production of high volumes of granulocyte precursors for use in treating cancer, thus overcoming problems of isolating sufficient quantities of fresh granulocytes (e.g. neutrophils) from a donor, which is a limitation of the current therapeutic methods.

[0542] Stem cells suitable for use in the methods of the invention may be selected on the basis of the cytocidal activity of a donor’s cells. Suitable ways in which CKA may be calculated have been described above. More detail of a suitable method is set out in Example 21 .

[0543] Suitably, such a screening procedure may be employed as an optional preliminary step in a method of the invention, such as a method employing the v0.3 (v0.3c) or v0.4 (optionally v0.4c or v0.4d) protocols disclosed herein.

[0544] Thus, in a suitable embodiment, a granulocyte precursor cell, or cell population, in accordance with the invention has been derived from a donor having a granulocyte (neutrophil) with high CKA.

[0545] Further, in a suitable embodiment, the population of stem cells employed in a method of the invention, such as a method of the thirteenth of fourteenth aspect of the invention, has been derived from a donor having a granulocyte (neutrophil) with high CKA.

[0546] The term “obtainable” as used herein also encompasses the term “obtained”.

[0547] The method of obtaining the granulocyte or precursor may comprise: culturing a population of stem cells in cell culture conditions to produce a population of granulocyte progenitor cells, wherein the cell culture conditions for producing the granulocyte precursor cells comprise the presence of:

[0548] SCF; • FLT-3 ligand;

[0549] • TPO; and

[0550] • a pyrimido-[4,5-b]-indole derivative that promotes the expansion of hematopoietic stem cells; to produce the population of granulocyte progenitor cells, and further comprising culturing granulocyte progenitor cells of this population in cell culture conditions that promote differentiation of the granulocyte progenitor cells into granulocytes, or precursors thereof, the cell culture conditions comprising the presence of:

[0551] • a basal medium;

[0552] • SCF;

[0553] • TPO; and

[0554] • G-CSF.

[0555] The following paragraphs provide guidance as to certain embodiments that may be used in respect of the culture conditions employed in the methods of obtaining the granulocyte, or precursor thereof. Cytokine concentrations provided may be based on the volume of media that is added and is not always a reflection of the concentration of the cytokine overall when in culture. This may be born in mind when considering the volumes of media used to ‘top up’ the culture. Alternatively, in the case of TNFa, the concentration may be based on the total volume of culture as this cytokine is added separately. References to concentrations of pyrimido-[4,5- b]-indole derivatives used may be based on the concentration of the compound overall when in culture.

[0556] The expansion medium may comprise Iscove’s Modified Dulbecco’s Medium (IMDM). The expansion step may last from day E0 to day E8.

[0557] The expansion steps employ required cytokines, and may further employ optional cytokines, as discussed further below.

[0558] Methods of the invention comprising an expansion step may utilise cell culture condition in which stem cell factor (SCF), FLT-3 ligand (FLT3-L) and thrombopoietic (TPO) are all present.

[0559] SCF may be provided at a concentration of approximately 0.01-10 pg / ml. SCF may be provided at a concentration of approximately 0.05-0.3 pg / ml. SCF may be provided at a concentration of approximately 0.1 -0.2 pg / ml. FLT-3 ligand may be provided at a concentration of approximately 0.01-10 pg / ml. FLT-3 ligand may be provided at a concentration of approximately 0.05-0.3 pg / ml. FLT-3 ligand may be provided at a concentration of approximately 0.1 -0.2 pg / ml.

[0560] TPO may be provided at a concentration of approximately 0.001-0.2 pg / ml. TPO may be provided at a concentration of approximately 0.005-0.03 pg / ml. TPO may be provided at a concentration of approximately 0.01-0.02 pg / ml.

[0561] An expansion phase to be employed in a method of the invention may optionally also comprise supplementation with the cytokines interleukin-3 (IL-3) and / or interleukin-6 (IL-6). Protocols in which supplementation with both IL-3 and IL-6 is employed are referred to at various points in this specification as protocol “v0.4c”. In contrast, protocols which omit supplementation with both IL-3 and IL-6 may be referred to in the specification as protocol “v0.4d”.

[0562] In the case that supplementation with IL-3 and / or IL-6 is to be used in an expansion step of the invention, IL-3 may be provided in the culture conditions at a concentration of approximately 0.0001-0.015 pg / ml. IL-6 may be provided in the culture conditions at a concentration of approximately 0.0001-0.015 pg / ml.

[0563] It may be preferred to employ a method of the invention in which the expansion phase is not supplemented with IL-3 or with IL-6 (e.g., the “v0.4d” protocol).

[0564] The expansion medium may comprise insulin-transferrin-selenium (ITS) and / or human serum albumin (HSA). The HSA may be recombinant HSA. HSA may be provided at a weight / volume (w / v) concentration of approximately 0.1% - 5%. For example, HAS may be provided as a supplement may be provided at a concentration of approximately 1%.

[0565] On the first day of expansion the medium to be used to generate progenitor cells may comprise IMDM with L-glutamine, 1 X ITS, 1% HSA, SCF (0.2 pg / ml), FLT-3L (0.2 pg / ml), TPO (0.02 pg / ml), IL-3 (0.015 pg / ml), and IL-6 (0.015 pg / ml). On the first day of expansion the medium may comprise IMDM with glutaMAX, 1 X ITS, 1% HSA, SCF (0.2 pg / ml), FLT-3L (0.2 pg / ml), TPO (0.02 pg / ml), IL-3 (0.015 pg / ml), and IL-6 (0.015 pg / ml). On the first day of expansion the medium to be used in a method in accordance with the first aspect of the invention may comprise IMDM with L-glutamine, 1 X ITS, 1% HSA, SCF (0.2 pg / ml), FLT-3L (0.2 pg / ml), and TPO (0.02 pg / ml). On the first day of expansion, the medium to be used may comprise IMDM with glutaMAX, 1 X ITS, 1% HSA, SCF (0.2 pg / ml), FLT-3L (0.2 pg / ml), and TPO (0.02 pg / ml).

[0566] A class of compounds known as pyrimido-[4,5-b]-indole derivatives are known to promote the expansion of hematopoietic stem cells, and to promote the retention of “sternness” in stem cells cultured in their presence. Examples of such compounds include the commercially available agents UM171 and UM729.

[0567] UM 171 represents a favoured example of a pyrimido-[4,5-b]-indole derivative that may be used in the methods of the invention. The pyrimido-[4,5-b]-indole derivative may be provided to the cells in one incidence, two incidences, three incidences, four incidences, or five incidences during the expansion phase. The pyrimido-[4,5-b]-indole derivative may be provided to the cells in four incidences during the expansion phase. In such an embodiment the incidences may suitably take place on E0, E4, E6 and E8. The pyrimido-[4,5-b]-indole derivative may provided to the cells in three incidences during the expansion phase. In such an embodiment the incidences may suitably take place on E0, E4 and E6.

[0568] The pyrimido-[4,5-b]-indole derivative may be UM 171 and may be provided to the cells during expansion in four incidences. Suitably these incidences may be at E0, E4, E6 and E8.

[0569] In one embodiment, the pyrimido-[4,5-b]-indole derivative is UM 171 , which is provided to the cells during expansion in three incidences. Suitably, the pyrimido-[4,5-b]-indole derivative is UM 171 , which may be provided to the cells during expansion in three incidences at E0, E4, and E6.

[0570] In one embodiment, the pyrimido-[4,5-b]-indole derivative is UM 171 , which is provided to the cells during expansion in three incidences at E1 , E4, and E6. For example, the UM 171 may be provided to the cells at a concentration of 50 nM at E1 , at a concentration of 25 nM at E4, and at a concentration of 50 nM at E6. The pyrimido-[4,5-b]-indole derivative may be UM729, which is provided to the cells during expansion in three incidences at E0, E4, and E6.

[0571] The pyrimido-[4,5-b]-indole derivative may be UM729 and may be provided to the cells during expansion in three incidences at E1 , E4, and E6. In a suitable example, the UM729 may be provided to the cells at a concentration of 500 nM at E1 , at a concentration of 50 nM at E4, and at a concentration of 1 pM at E6. Suitably, the pyrimido-[4,5-b]-indole derivative may be provided at a concentration within the range of 15 nM - 1.5pM.

[0572] In a suitable embodiment, the pyrimido-[4,5-b]-indole derivative (such as UM 171 or UM729) may be provided at a concentration of approximately 50nM - 1 M. For example, the pyrimido- [4,5-b]-indole derivative may be provided at a concentration of approximately 50 nM, approximately 100 nM, approximately 150 nM, approximately 200 nM, approximately 250 nM, approximately 300 nM, approximately 350 nM, approximately 400 nM, approximately 450 nM, approximately 500 nM, approximately 550 nM, approximately 600 nM, approximately 650 nM, approximately 700 nM, approximately 750 nM, approximately 800 nM, approximately 850 nM, approximately 900 nM, approximately 950 nM, approximately 1 pM, approximately 1.25 pM, approximately 1.5 pM.

[0573] Optionally, the cell culture conditions may further comprise: ITS; and / or HSA.

[0574] Similarly, the cell culture conditions may comprise one or more conditions independently selected from the group consisting of: SCF at a concentration of approximately 0.2 pg / ml; FLT- 3 ligand at a concentration of approximately 0.2 pg / ml; TPO at a concentration of approximately 0.2 pg / ml; and HSA at a concentration of approximately 1%.

[0575] Methods of the invention may comprise a differentiation step, in which cells are cultured in conditions the promote differentiation of a population of progenitor cells into a population of granulocytes, or precursors thereof. Suitably the cells produced may be neutrophils (or precursors thereof).

[0576] In a suitable embodiment, a differentiation step lasts from day DO to day D4. In a method comprising an expansion step, E8 of the expansion step may correspond to DO of the differentiation step.

[0577] The basal medium employed in a differentiation step in accordance with the invention may suitably be Stemline II. In an alternative embodiment, the basal medium employed in a differentiation step in accordance with the invention may suitably be IMDM. Alternatively, in one embodiment, the basal medium is not IMDM.

[0578] In a suitable embodiment of a differentiation step one, two or three of each of SCF, TPO, and G-CSF provided in a first incidence of administration may be provided in an amount of between approximately 0.111 ng and 0.555 ng per 1000 cells, for example an amount of between approximately 0.222 ng and 0.444 ng per 1000 cells, such as approximately 0.333 ng per 1000 cells in culture.

[0579] In a suitable embodiment of a differentiation step one, two or three of each of SCF, TPO, and G-CSF provided in a second incidence of administration may be provided in an amount approximately twice that provided in the first incidence of administration. For example, one, two or three of each of SCF, TPO, and G-CSF provided in a second incidence of administration may be provided in an amount of between approximately 0.444 ng and 0.888 ng per 1000 cells, for example an amount of between approximately 0.555 ng and 0.777 ng per 1000 cells, such as approximately 0.666 ng per 1000 cells in culture.

[0580] A differentiation step suitable for use in the methods of the invention may also utilise cell culture conditions that comprise supplementation with one, two or three cytokines selected from the group consisting of: granulocyte-macrophage colony-stimulation factor (GM-CSF), interleukin- 3 (IL-3) and tumour necrosis factor alpha (TNFa).

[0581] For example, a differentiation step suitable for use in the methods of the invention may make use of cell culture conditions that comprise one, two or three of the following:

[0582] • GM-CSF at a concentration of approximately 0.001 - 0.03 pg / ml;

[0583] • IL-3 at a concentration of approximately 0.05 - 0.2 pg / ml; and

[0584] • TNFa at a concentration of approximately 0.0001 - 0.005 pg / ml.

[0585] Suitably, a differentiation step suitable for use in the methods of the invention may make use of cell culture conditions that comprise one, two or three of the following:

[0586] • GM-CSF at a concentration of approximately 0.01 pg / ml;

[0587] • IL-3 at a concentration of approximately 0.13 pg / ml; and

[0588] • TNFa at a concentration of approximately 0.001 pg / ml.

[0589] In a suitable embodiment, cell culture conditions used in a differentiation step employed in a method of the invention may comprise each of the following:

[0590] • GM-CSF at a concentration of approximately 0.01 pg / ml; and

[0591] • IL-3 at a concentration of approximately 0.13 pg / ml; and

[0592] • TNFa at a concentration of approximately 0.001 pg / ml. The methods may result in the production of populations of progenitor cells. The populations of progenitor cells may also be employed in the methods of obtaining the granulocyte, or precursor thereof.

[0593] In a suitable embodiment of a population of progenitor cells:

[0594] • at least 50% of the cells of the population may express CD34;

[0595] • at least 50% of the cells of the population may express CD13;

[0596] • at least 50% of the cells of the population may express HLA-DR;

[0597] • at least 50% of the cells of the population may express CD49d;

[0598] • at least 50% of the cells of the population may express CD33;

[0599] • less than 50% of the cells of the population may express CD15;

[0600] • less than 50% of the cells of the population may express CD38; and

[0601] • less than 50% of the cells of the population may express CD45RA.

[0602] By way of example, in a suitable embodiment of a population of progenitor cells:

[0603] • between 80% and 90% of the cells of the population may express CD34; and / or

[0604] • between 90% and 100% of the cells of the population may express CD13; and / or

[0605] • between 70% and 85% of the cells of the population may express HLA-DR; and / or

[0606] • between 95% and 100% of the cells of the population may express CD49d; and / or

[0607] • between 95% and 10% of the cells of the population may express CD33; and / or

[0608] • between 0% and 10% of the cells of the population may express CD15; and / or

[0609] • between 20% and 50% of the cells of the population may express CD38; and / or

[0610] • between 15% and 30% of the cells of the population may express CD45RA.

[0611] Suitably, in a population of progenitor cells:

[0612] • between 80% and 90% of the cells of the population may express CD34;

[0613] • between 90% and 100% of the cells of the population may express CD13;

[0614] • between 70% and 85% of the cells of the population may express HLA-DR;

[0615] • between 95% and 100% of the cells of the population may express CD49d;

[0616] • between 95% and 10% of the cells of the population may express CD33;

[0617] • between 0% and 10% of the cells of the population may express CD15;

[0618] • between 20% and 50% of the cells of the population may express CD38; and

[0619] • between 15% and 30% of the cells of the population may express CD45RA. The inventors have also identified methods by which granulocytes or precursors thereof may be primed, in order to amplify properties of the cells that increase their therapeutic utility. In particular, priming of the granulocytes or precursors thereof by such methods may amplify their cytocidal activity in a manner that may increase their therapeutic utility.

[0620] Accordingly, the method of priming may comprise culturing the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) in the presence of GM-CSF, and optionally one or more cytokines selected from the group consisting of: TNF, IFN-a, IFN-p, IL-15, and IL-18.

[0621] The method of priming the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may optionally comprise a further step of purifying the primed granulocyte or precursor thereof (e.g., a granulocyte precursor cell) produced.

[0622] GM-CSF may be used in cell culture conditions for a priming step at a concentration of 1-1000 ng / mL, 2-500 ng / mL, 3-250 ng / mL, 4-200 ng / mL. GM-CSF may be used at a concentration of 5-150 ng / mL, for example at a concentration of 10-130 ng / mL.

[0623] TNF may be used in cell culture conditions for a priming step at a concentration of 0.001-10 ng / mL, 0.002-5 ng / mL, 0.003-2.5 ng / mL, 0.004-2 ng / mL. TNF may be used at a concentration of 0.005-1 .5 ng / mL, for example at a concentration of 0.01-1 ng / mL.

[0624] IFN-a may be used in cell culture conditions for a priming step at a concentration of 1-100 ng / mL, 2-50 ng / mL, 3-25 ng / mL, 4-20 ng / mL. IFN-a may be used at a concentration of 5-15 ng / mL, for example at a concentration of 10 ng / mL.

[0625] IFN-p may be used in cell culture conditions for a priming step at a concentration of 1-100 ng / mL, 2-50 ng / mL, 3-25 ng / mL, 4-20 ng / mL. IFN-p may be used at a concentration of 5-15 ng / mL, for example at a concentration of 10 ng / mL.

[0626] IL-15 may be used in cell culture conditions for a priming step at a concentration of 1-100 ng / mL, 2-50 ng / mL, 3-25 ng / mL, 4-20 ng / mL. IL-15 may be used at a concentration of 5-15 ng / mL, for example at a concentration of 10 ng / mL.

[0627] IL-18 may be used in cell culture conditions for a priming step at a concentration of 1-100 ng / mL, 2-50 ng / mL, 3-25 ng / mL, 4-20 ng / mL. IL-18 may be used at a concentration of 5-15 ng / mL, for example at a concentration of 10 ng / mL. IL-3 may be used in cell culture conditions for a priming step at a concentration of 1-1000 ng / mL, 2-500 ng / mL, 3-250 ng / mL, 4-200 ng / mL. IL-3 may be used at a concentration of 5- 150 ng / mL, for example at a concentration of 10-130 ng / mL.

[0628] Priming may involve culturing the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) in the presence of GM-CSF at a concentration of approximately 130ng / mL, and optionally one or more cytokines selected from the group consisting of: TNF at a concentration of approximately 0.01-1.0ng / mL, IFN-a at a concentration of approximately 10ng / mL, IFN-p at a concentration of approximately 10ng / mL, IL-15 at a concentration of approximately 10ng / mL, IL-18 at a concentration of approximately 10ng / mL, and IL-3 at a concentration of approximately 130ng / mL.

[0629] Cells undergoing priming may be cultured in the presence of GM-CSF, G-CSF, SCF, TPO, and IL-15. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 10ng / mL, G-CSF at a concentration of approximately 130ng / mL, SCF at a concentration of approximately 130ng / mL, TPO at a concentration of approximately 130ng / mL, and IL-15 at a concentration of approximately 10ng / mL.

[0630] Cells undergoing priming may be cultured in the presence of GM-CSF, G-CSF, SCF, TPO, and TNF. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 100ng / mL, G-CSF at a concentration of approximately 130ng / mL, SCF at a concentration of approximately 130ng / mL, TPO at a concentration of approximately 130ng / mL, and TNF at a concentration of approximately 10ng / mL.

[0631] Cells undergoing priming may be cultured in the presence of GM-CSF and IL-3. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 130ng / mL and IL-3 at a concentration of approximately 130ng / mL.

[0632] Cells undergoing priming may be cultured in the presence of GM-CSF and IL-15. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 130ng / mL and IL-15 at a concentration of approximately 10ng / mL. Cells undergoing priming may be cultured in the presence of GM-CSF and IL-18. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 130ng / mL and IL-18 at a concentration of approximately 10ng / mL.

[0633] Cells undergoing priming may be cultured in the presence of GM-CSF and IL-16. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 130ng / mL and IL-16 at a concentration of approximately 10ng / mL.

[0634] Cells undergoing priming may be cultured in the presence of GM-CSF and TNF. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 130ng / mL and TNF at a concentration of approximately 1 ng / mL.

[0635] Cells undergoing priming may be cultured in the presence of GM-CSF, G-CSF, SCF, TPO, and IFN-a. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be cultured in the presence of GM-CSF at a concentration of approximately 130ng / mL, G-CSF at a concentration of approximately 130ng / mL, SCF at a concentration of approximately 130ng / mL, TPO at a concentration of approximately 130ng / mL, and IFN-a at a concentration of approximately 10ng / mL.

[0636] The priming step may last any suitable period of time. For example, the priming step may be last for 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 18 hours, 24 hours, 30 hours, 36 hours, 42 hours, 48 hours, 54 hours, 60 hours, 72 hours, 78 hours, 84 hours, 90 hours, or 96 hours. The priming step may last for 1-96 hours, 2-90 hours, 3-84 hours, 6-78 hours, 12-72 hours, 18-54 hours, or 24-48 hours. The priming may comprise culture incorporating the cytokines discussed above, for example at the concentrations set out above, for a period of one, two or three days. In particular, the priming may comprise culture incorporating the priming cytokine combinations referred to for two days.

[0637] A priming step may be incorporated at any appropriate stage. That said, priming may typically occur during the period in which the progenitor cells are cultured in conditions that promote differentiation of the progenitor cells into the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . For example, priming may begin on the first day of culture of the progenitor cells, the second day of culture of the progenitor cells, the third day of culture of the progenitor cells, the fourth day of culture of the progenitor cells, or on the fifth day of culture of the progenitor cells in conditions that promote their differentiation into the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0638] A priming step may occur after the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) has been produced, and optionally after the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) has been harvested. For example, priming may occur before or after cryopreservation of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) in accordance with the invention.

[0639] In the case of priming steps practiced for two days, the priming may take place on days 3 and

[0640] 4 of the culture conditions that promote differentiation of the progenitor cells into the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , on days 4 and 5 of such culture, or on days 5 and 6 of such culture. For the avoidance of doubt, any of the priming protocols described above may suitably be practiced on days 3 and 4, days 4 and 5, or days

[0641] 5 and 6 of the culture conditions that promote differentiation of progenitor cells into the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0642] The antibody or antigen-binding protein described herein is suitable for binding to the ADCC- promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . By way of said binding, the antibody or antigen-binding protein described herein may promote ADCC. The term “promote” as used in this context means that the antibody or antigen-binding protein may activate the FcR and induce or increase the cytotoxic activity of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Thus, the term “promote” as used in this context encompasses “induce”. Accordingly, the antibody or antigen-binding protein described herein may bind to the ADCC-promoting FcR and induce ADCC.

[0643] The antibody or antigen-binding protein may be any suitable antibody or antigen-binding protein. The antibody or antigen-binding protein comprises a domain for binding to an ADCC- promoting FcR on a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) described herein. The domain for binding to an ADCC-promoting FcR on a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) described herein may be capable of binding to the ADCC-promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and activating the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Preferably, the domain for binding to an ADCC-promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is a human domain.

[0644] The domain for binding to an ADCC-promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be an Fc domain. Accordingly, the antibody or antigenbinding protein may comprise an Fc domain. As set out elsewhere, the Fc domain may be a region of an antibody or antigen-binding protein that binds to an Fc receptor and activates the cell on which the Fc receptor is present. Preferably, the domain may be a human Fc domain. Preferably, the Fc domain may be an Fc-enabled domain. The Fc-mediated function of an antibody or antigen-binding protein may be avoided by introducing mutations into the Fc domain to eliminate binding of the Fc domain to an Fc receptor, or to eliminate activation of the cell to which the Fc domain binds. Accordingly, the Fc-enabled domain may be an Fc domain which is capable of binding to an Fc receptor (e.g., the ADCC-promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ) and activating the cell on which the Fc receptor is present (e.g., the granulocyte or precursorthereof (e.g., a granulocyte precursor cell) ).

[0645] The Fc domain may be an IgG, IgA, IgM, IgD, or IgE Fc domain. Preferably, the Fc domain may be an IgG or an IgA Fc domain. The antibody or antigen-binding protein may comprise an IgG Fc domain. The IgG Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to any one of SEQ ID NOs: 1-4. The IgG Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 1. The IgG Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 2. The IgG Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 3. The IgG Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 4. Preferably, the IgG Fc domain may have the amino acid sequence of any one of SEQ ID NOs: 1-4. The IgG Fc domain may have the amino acid sequence of SEQ ID NO: 1 . The IgG Fc domain may have the amino acid sequence of SEQ ID NO: 2. The IgG Fc domain may have the amino acid sequence of SEQ ID NO: 3. The IgG Fc domain may have the amino acid sequence of SEQ ID NO: 4.

[0646] The antibody or antigen-binding protein may comprise an IgA Fc domain. The IgA Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to any one of SEQ ID NO: 5. Preferably, the IgA Fc domain may have the amino acid sequence of SEQ ID NO: 5.

[0647] Where the antibody or antigen-binding protein comprises an IgG Fc domain, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may comprise an ADCC-promoting IgG FcR. For example, the antibody or antigen-binding protein may comprise an IgG Fc domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD32 and / or CD64. Preferably, the antibody or antigen-binding domain may comprise an IgG Fc domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD64. The antibody or antigen-binding domain may comprise an IgG Fc domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD32.

[0648] The IgG Fc domain may be an lgG1 Fc domain, an lgG2 Fc domain, an lgG3 Fc domain, or an lgG4 Fc domain. The IgG Fc domain may be an IgG 1 Fc domain. The IgG 1 Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 1. Preferably, the lgG1 Fc domain may have the amino acid sequence of SEQ ID NO: 1. The IgG Fc domain may be an lgG2 Fc domain. The lgG2 Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 2. Preferably, the lgG2 Fc domain may have the amino acid sequence of SEQ ID NO: 2. The IgG Fc domain may be an lgG3 Fc domain. The lgG3 Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 3. Preferably, the lgG3 Fc domain may have the amino acid sequence of SEQ ID NO: 3. The IgG Fc domain may be an lgG4 Fc domain. The lgG4 Fc domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 4. Preferably, the lgG4 Fc domain may have the amino acid sequence of SEQ ID NO: 4.

[0649] Where the antibody or antigen-binding protein comprises an IgA Fc domain, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may comprise an ADCC-promoting IgA FcR. Preferably, the antibody or antigen-binding domain may comprise an IgA Fc domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD89.

[0650] The domain for binding to an ADCC-promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be a constant domain. Preferably, the constant domain may be a human constant domain. The antibody or antigen-binding protein may comprise an IgG, IgA, IgM, IgD, or IgE constant domain. For example, the antibody or antigen-binding protein may comprise an IgG or an IgA constant domain. Preferably, the antibody or antigenbinding protein may comprise an IgG constant domain. The IgG constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to any one of SEQ ID NOs: 6-9. The IgG constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 6. The IgG constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 7. The IgG constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 8. The IgG constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 9. Preferably, the IgG constant domain may have the amino acid sequence of any one of SEQ ID NOs: 6-9. The IgG constant domain may have the amino acid sequence of SEQ ID NO: 6. The IgG constant domain may have the amino acid sequence of SEQ ID NO: 7. The IgG constant domain may have the amino acid sequence of SEQ ID NO: 8. The IgG constant domain may have the amino acid sequence of SEQ ID NO: 9.

[0651] The antibody or antigen-binding protein may comprise an IgA constant domain. The IgA constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to any one of SEQ ID NO 10. Preferably, the IgA constant domain may have the amino acid sequence of SEQ ID NO 10.

[0652] Where the antibody or antigen-binding protein comprises an IgG constant domain, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may comprise an ADCC- promoting IgG FcR. For example, the antibody or antigen-binding protein may comprise an IgG constant domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD32 and / or CD64. Preferably, the antibody or antigen-binding domain may comprise an IgG constant domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD64. The antibody or antigen-binding domain may comprise an IgG constant domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD32.

[0653] The IgG constant domain may be an IgG 1 constant domain, an lgG2 constant domain, an lgG3 constant domain, or an lgG4 constant domain. The IgG constant domain may be an IgG 1 constant domain. The lgG1 constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 6. Preferably, the IgG 1 constant domain may have the amino acid sequence of SEQ ID NO: 6. The IgG constant domain may be an lgG2 constant domain. The lgG2 constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 7. Preferably, the lgG2 constant domain may have the amino acid sequence of SEQ ID NO: 7. The IgG constant domain may be an lgG3 constant domain. The lgG3 constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 8. Preferably, the lgG3 constant domain may have the amino acid sequence of SEQ ID NO: 8. The IgG constant domain may be an lgG4 constant domain. The lgG4 constant domain may have an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 99% or 100% identity to SEQ ID NO: 9. Preferably, the lgG4 constant domain may have the amino acid sequence of SEQ ID NO: 9.

[0654] Where the antibody or antigen-binding protein comprises an IgA constant domain, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may comprise an ADCC- promoting IgA FcR. Preferably, the antibody or antigen-binding domain may comprise an IgA constant domain and the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may express CD89.

[0655] The antibody or antigen-binding protein may be suitable for binding to an antigen. The antibody or antigen-binding protein may bind to an antigen. The antibody or antigen-binding protein may be suitable for specifically binding to an antigen. The antibody or antigen-binding protein may specifically bind to an antigen. Preferably, the antibody or antigen-binding protein may be suitable for binding to an antigen on a target. The antibody or antigen-binding protein may bind to an antigen on a target. The antibody or antigen-binding protein may be suitable for specifically binding to an antigen on a target. The antibody or antigen-binding protein may specifically bind to an antigen on a target.

[0656] The antigen may be any suitable antigen. For example, the antigen may be associated with the disorder being treated. The antigen may be causative of the disorder being treated.

[0657] The antigen may be selected from the group comprising or consisting of: AFP, AKAP-4, ALK, alpha-fetoprotein, Androgen receptor, B7H3, BAGE, BCA225, BCAA, Bcr-abl, beta-Catenin, beta-HCG, beta-human chorionic gonadotropin, BORIS, BTAA, CA 125, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72-4, CA-50, CAM 17.1 , CAM43, Carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33 / IL3Ra, CD68\P1 , CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG4), c-Met, CO-029, CSPG4, Cyclin B1 , cyclophilin C-associated protein, CYP1B1 , E2A-PRL, EGFR, EGFRvlll, ELF2M, EpCAM, EphA2, EphrinB2, Epstein Barr virus antigens EBVA, ERG (TMPRSS2ETS fusion gene), ETV6-AML, FAP, FGF-5, Fos-related antigen 1 , Fucosyl GM1 , G250, Ga733\EpCAM, GAGE-1 , GAGE-2, GD2, GD3, glioma- associated antigen, GloboH, Glycolipid F77, GM3, GP 100, GP 100 (Pmel 17), H4-RET, HER- 2, HER-2 / neu, HER-2 / Neu / ErbB-2, high-molecular-weight melanoma-associated antigen (HMW-MAA), HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, Idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin growth factor (IGF)-I, intestinal carboxyl esterase, K-ras, LAGE-1a, LCK, lectin-reactive AFP, Legumain, LMP2, M344, MA-50, Mac-2 binding protein, MAD-CT-1 , MAD-CT-2, MAGE, MAGE A1 , MAGE A3, MAGE-1 , MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART-1 , MART-1 / MelanA, M-CSF, melanoma-associated chondroitin sulfate proteoglycan (MCSP), Mesothelin, MG7-Ag, ML-IAP, MN-CA IX, MOV18, MUC1 , Mum-1 , hsp70-2, MYCN, MYL-RAR, NA17, NB / 70K, neuron-glial antigen 2 (NG2), nm- 23H1 , NuMa, NY-BR-1 , NY-CO-1 , NY-ESO, NY-ESO-1 , NY-ESO-1 , OY-TES1 , p15, p16, p180erbB3, p185erbB2, p53, p53 mutant, PAX3, PAX5, PDGFR-beta, PLAC1 , Polysialic Acid, prostate-carcinoma tumor antigen-1 (PCTA-1), prostate-specific antigen, prostatic acid phosphatase (PAP), Proteinase3 (PR1), PSA, PSCA, PSMA, RAGE-1 , Ras, Ras-mutant, RCAS1 , RGS5, RhoC, ROR1 , RU1 , RU2 (AS), SART3, SDCCAG16, sLe(a), Sperm protein 17, SSX2, STn, Survivin, TA-90, TAAL6, a TAG-72, telomerase, thyroglobulin, Tie 2, TLP, Tn, TPS, Trop-2, TRP-1 , TRP-2, TRP-2, TSP-180, Tyrosinase, VEGF, VEGFR2, VISTA, WT1 , XAGE 1 , 43-9F, 5T4, and 791Tgp72; CD20, CD19, PD-L1 , CD38, SLAMF7, CCR4, DLL3, FLT3, STEAP1 , MUC17, PD-1 , GPCR5D, BCMA, CDTLA-4, LAG-3, TIM3, TIGIT, CTLA4, CD27, TGF-beta, CD28, PD-L1 , HER3, HER2 (ECD2), HER2 (ECD4), CD47, IMCgpl 00, CD3, PRAME, MSLN, V delta 2, MUC16, CD30, CD16A, MAGE A4, MAGE A8, CLL4, and VEGF- A.

[0658] The antigen may be selected from the group comprising or consisting of: CD52, PD-L1 , EGFR, CD38, GD2, SLAMF7, CD38, HER-2, CCR4, CD20, CD19, MET, PD-1 , CTLA-4, and LAG-3.

[0659] The antigen may be a cancer antigen. The cancer antigen may be selected from the group comprising or consisting of: AFP, AKAP-4, ALK, alpha-fetoprotein, Androgen receptor, B7H3, BAGE, BCA225, BCAA, Bcr-abl, beta-Catenin, beta-HCG, beta-human chorionic gonadotropin, BORIS, BTAA, CA 125, CA 15-3, CA 195, CA 19-9, CA 242, CA 27.29, CA 72- 4, CA-50, CAM 17.1 , CAM43, Carbonic anhydrase IX, carcinoembryonic antigen, CD22, CD33 / IL3Ra, CD68\P1 , CDK4, CEA, chondroitin sulfate proteoglycan 4 (CSPG4), c-Met, CO- 029, CSPG4, Cyclin B1 , cyclophilin C-associated protein, CYP1 B1 , E2A-PRL, EGFR, EGFRvlll, ELF2M, EpCAM, EphA2, EphrinB2, Epstein Barr virus antigens EBVA, ERG (TMPRSS2ETS fusion gene), ETV6-AML, FAP, FGF-5, Fos-related antigen 1 , Fucosyl GM1 , G250, Ga733\EpCAM, GAGE-1 , GAGE-2, GD2, GD3, glioma-associated antigen, GloboH, Glycolipid F77, GM3, GP 100, GP 100 (Pmel 17), H4-RET, HER-2 / neu, HER-2 / Neu / ErbB-2, high-molecular-weight melanoma-associated antigen (HMW-MAA), HPV E6, HPV E7, hTERT, HTgp-175, human telomerase reverse transcriptase, Idiotype, IGF-I receptor, IGF-II, IGH-IGK, insulin growth factor (IGF)-I, intestinal carboxyl esterase, K-ras, LAGE-1 a, LCK, lectin-reactive AFP, Legumain, LMP2, M344, MA-50, Mac-2 binding protein, MAD-CT-1 , MAD-CT-2, MAGE, MAGE A1 , MAGE A3, MAGE-1 , MAGE-3, MAGE-4, MAGE-5, MAGE-6, MART-1 , MART- 1 / MelanA, M-CSF, melanoma-associated chondroitin sulfate proteoglycan (MCSP), Mesothelin, MG7-Ag, ML-IAP, MN-CA IX, MOV18, MUC1 , Mum-1 , hsp70-2, MYCN, MYL- RAR, NA17, NB / 70K, neuron-glial antigen 2 (NG2), nm-23H1 , NuMa, NY-BR-1 , NY-CO-1 , NY- ESO, NY-ESO-1 , NY-ESO-1 , OY-TES1 , p15, p16, p180erbB3, p185erbB2, p53, p53 mutant, PAX3, PAX5, PDGFR-beta, PLAC1 , Polysialic Acid, prostate-carcinoma tumor antigen-1 (PCTA-1), prostate-specific antigen, prostatic acid phosphatase (PAP), Proteinase3 (PR1), PSA, PSCA, PSMA, RAGE-1 , Ras, Ras-mutant, RCAS1 , RGS5, RhoC, ROR1 , RU1 , RU2 (AS), SART3, SDCCAG16, sLe(a), Sperm protein 17, SSX2, STn, Survivin, TA-90, TAAL6, a TAG-72, telomerase, thyroglobulin, Tie 2, TLP, Tn, TPS, Trop-2, TRP-1 , TRP-2, TRP-2, TSP- 180, Tyrosinase, VEGF, VEGFR2, VISTA, WT1 , XAGE 1 , 43-9F, 5T4, and 791Tgp72; CD20, CD19, PD-L1 , CD38, SLAMF7, and CCR4.

[0660] The cancer antigen may be a tumour-associated antigen or a tumour-specific antigen. The tumour-associated antigen may be selected from the group comprising or consisting of: CD52, PD-L1 , EGFR, CD38, GD2, SLAMF7, CD38, HER-2, CCR4, CD20, and CD19.

[0661] The antigen may be a checkpoint protein antigen. The checkpoint protein antigen may be selected from the group comprising or consisting of: PD-1 (CD279), PD-L1 (CD274), CTLA4 (CD152), LAG-3 (CD223), TIM-3 (CD366), TIGIT, B7-H3 (CD276), B7-H4 (VTCN1), NKG2A, and CD73. For example, the checkpoint protein may be selected from the group comprising or consisting of: PD-1 (CD279), PD-L1 (CD274), CTLA4 (CD152), and LAG-3 (CD223).

[0662] A cancer may be a primary cancer. A cancer may be a metastatic cancer.

[0663] A cancer may be of one or more of: head and neck cancer, cervical cancer (such as metastatic cervical cancer), pancreatic cancer, liver cancer, oesophageal cancer, stomach cancer, ovarian cancer, lung cancer, bladder cancer, kidney cancer, brain cancer, prostate cancer, myeloma cancer, non-Hodgkin’s lymphoma (NHL), larynx cancer, uterine cancer, or breast cancer. Suitably a cancer is head and neck cancer, such as a head and neck squamous cell carcinoma (HNSCC). Suitably a cancer is cervical cancer, such as a metastatic cervical cancer.

[0664] A target may be a primary cancer cell. A target may be a metastatic cancer cell.

[0665] A target may be of one or more of: a head and neck cancer cell, a cervical cancer cell (such as metastatic cervical cancer cell), a pancreatic cancer cell, a liver cancer cell, an oesophageal cancer cell, a stomach cancer cell, an ovarian cancer cell, a lung cancer cell, a bladder cancer cell, a kidney cancer cell, a brain cancer cell, a prostate cancer cell, a myeloma cancer cell, a non-Hodgkin’s lymphoma (NHL) cell, a larynx cancer cell, a uterine cancer cell, or a breast cancer cell. Suitably a target is a head and neck cancer cell, such as a head and neck squamous cell carcinoma (HNSCC) cell. Suitably a target is a cervical cancer cell, such as a metastatic cervical cancer cell.

[0666] Suitably the antibody (or antigen binding protein) binds an antigen associated with a head and neck cancer cell, a cervical cancer cell (such as metastatic cervical cancer cell), a pancreatic cancer cell, a liver cancer cell, an oesophageal cancer cell, a stomach cancer cell, an ovarian cancer cell, a lung cancer cell, a bladder cancer cell, a kidney cancer cell, a brain cancer cell, a prostate cancer cell, a myeloma cancer cell, a non-Hodgkin’s lymphoma (NHL) cell, a larynx cancer cell, a uterine cancer cell, or a breast cancer cell.

[0667] Suitably the antibody (or antigen binding protein) binds an antigen associated with a head and neck cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a cervical cancer cell (such as metastatic cervical cancer cell). Suitably the antibody (or antigen binding protein) binds an antigen associated with a pancreatic cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a liver cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with an oesophageal cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a stomach cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with an ovarian cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a lung cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a bladder cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a kidney cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a brain cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a prostate cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a myeloma cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a non-Hodgkin’s lymphoma (NHL) cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a larynx cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a uterine cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a breast cancer cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a head and neck cancer cell, such as a head and neck squamous cell carcinoma (HNSCC) cell. Suitably the antibody (or antigen binding protein) binds an antigen associated with a cervical cancer cell, such as a metastatic cervical cancer cell.

[0668] The antigen may be expressed on a target. The target may be a target associated with the disorder being treated, or a target causative of the disorder being treated. The target may be associated with the disorder being treated. The target may be causative of the disorder being treated. The target may be a cancer cell or a pathogen. The target may be a pathogen, e.g., a bacterium, a fungus, a virus, or a macroparasite.

[0669] The antigen may be expressed on the surface of the target. The antigen may be expressed within the target.

[0670] The target may be a target cell. The target cell may be any suitable cell. For example, the target cell may be a disordered cell, a cell associated with the disorder being treated, or a cell causative of the disorder being treated. The target cell may be a disordered cell. The target cell may be associated with the disorder being treated. The target cell may be causative of the disorder being treated.

[0671] The target cell may be a fibroblast, a breast cell, a myofibroblast, an epithelial cell, a glial cell, a fat cell, an immune cell, a vascular cell, or a smooth muscle cell. Preferably, the target cell may be a cancer cell or a pathogen cell. The target cell may be a cancer cell. For example, the cancer cell may be a tumour cell. The cancer cell (e.g. tumour cell) may be a fibroblast, a breast cell, a myofibroblast, an epithelial cell, a glial cell, a fat cell, an immune cell, a vascular cell, or a smooth muscle cell. The cancer cell (e.g. tumour cell) may be a breast cell, i.e., a breast cancer cell. The target cell may be a pathogen cell, e.g., a bacterial cell, a fungal cell, or a macroparasite cell. The antigen may be expressed on the cell surface of the target cell. The antigen may be expressed within the target cell.

[0672] The antibody or antigen-binding protein may be a monoclonal or polyclonal antibody or antigen-binding protein. Preferably, the antibody or antigen-binding protein may be a monoclonal antibody or antigen-binding protein. The antibody or antigen-binding protein may be a polyclonal antibody or antigen-binding protein.

[0673] The antibody or antigen-binding protein may be a monospecific or multispecific antibody or antigen-binding protein. As used herein, a “multispecific” antibody or antigen-binding protein may refer to an antibody or antigen-binding protein which binds to more than one antigen. The antibody or antigen-binding protein may be a monospecific antibody or antigen-binding protein. The antibody or antigen-binding protein may be a multispecific antibody or antigen-binding protein.

[0674] The antibody or antigen-binding protein may comprise a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the HCDR1 , HCDR2, HCDR3, LCDR1 , LCDR2, and LCDR3 are the HCDR1 , HCDR2, HCDR3, LCDR1 , LCDR2, and LCDR3 of alemtuzumab, amivantamab, atezolizumab, avelumab, cemiplimab, cetuximab, daratumumab, dinutuximab, durvalumab, elotuzumab, ipilimumab, isatuximab, margetuximab, mogamulizumab, naxitamab, necitumumab, nivolumab, obinutuzumab, ofatumumab, pembrolizumab, pertuzumab, relatlimab, rituximab, tafasitamab, or trastuzumab, or a biosimilar thereof. The antibody or antigen-binding protein may comprise a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the HCDR1 , HCDR2, HCDR3, LCDR1 , LCDR2, and LCDR3 are the HCDR1 , HCDR2, HCDR3, LCDR1 , LCDR2, and LCDR3 of trastuzumab or a biosimilar thereof.

[0675] The antibody or antigen-binding protein may comprise a VH region and VL region amino acid sequence comprising the VH region and VL region amino acid sequence of alemtuzumab, amivantamab, atezolizumab, avelumab, cemiplimab, cetuximab, dratumumab, dinutuximab, durvalumab, elotuzumab, ipilimumab, isatuximab, margetuximab, mogamulizumab, naxitamab, necitumumab, nivolumab, obinutuzumab, ofatumumab, pembrolizumab, pertuzumab, relatlimab, rituximab, tafasitamab, or trastuzumab, or a biosimilar thereof. The antibody or antigen-binding protein may comprise a VH region and VL region amino acid sequence comprising the VH region and VL region amino acid sequence of trastuzumab or a biosimilar thereof.

[0676] The antibody or antigen-binding protein may bind to EGFR. The antibody or antigen-binding protein may bind to EGFR and comprise a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 11 , HCDR2 of SEQ ID NO: 12 and HCDR3 of SEQ ID NO: 13, and wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 14, LCDR2 of SEQ ID NO: 15 and LCDR3 of SEQ ID NO: 16.

[0677] The antibody or antigen-binding protein may bind to EGFR and comprise: a heavy chain CDR1 having 1 , 2 or 3 amino acid differences when compared to the HCDR1 of SEQ ID NO: 11 ; a heavy chain CDR2 having 1 , 2 or 3 amino acid differences when compared to the HCDR2 of SEQ ID NO: 12; a heavy chain CDR3 having 1 , 2 or 3 amino acid differences when compared to the HCDR3 of SEQ ID NO: 13; a light chain CDR1 having 1 , 2 or 3 amino acid differences when compared to the LCDR1 of SEQ ID NO: 14; a light chain CDR2 having 1 , 2 or 3 amino acid differences when compared to the LCDR2 of SEQ ID NO: 15; and / or (e.g., and) a light chain CDR3 having 1 , 2 or 3 amino acid differences when compared to the LCDR3 of SEQ ID NO: 16.

[0678] The antibody or antigen-binding protein may bind to EGFR and comprise a VH region amino acid sequence comprising SEQ ID NO: 17 and a VL region amino acid sequence comprising SEQ ID NO: 18.

[0679] The antibody or antigen-binding protein may bind to EGFR and comprise a VH region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 17. The antibody or antigen-binding protein may bind to EGFR and comprise a VL region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. The CDRs of such an antibody or antigen-binding protein may be identical to the CDRs of an antibody or antigen-binding protein comprising a VH region amino acid sequence comprising SEQ ID NO: 17 and a VL region amino acid sequence comprising SEQ ID NO: 18. Thus, the antibody or antigen binding protein may bind to EGFR and comprise: a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 11 , HCDR2 of SEQ ID NO: 12 and HCDR3 of SEQ ID NO: 13, and wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 14, LCDR2 of SEQ ID NO: 15 and LCDR3 of SEQ ID NO: 16, and wherein the antibody or antigen-binding protein comprises a VH region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 17, and a VL region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18.

[0680] The antibody or antigen-binding protein may bind to EGFR and comprise a heavy chain amino acid sequence comprising SEQ ID NO: 19 and a light chain amino acid sequence comprising SEQ ID NO: 20.

[0681] The antibody or antigen-binding protein may bind to EGFR and comprise a heavy chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 19. The antibody or antigen-binding protein may bind to EGFR and comprise a light chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20. The CDRs of such an antibody or antigen-binding protein may be identical to the CDRs of an antibody or antigen-binding protein comprising a heavy chain amino acid sequence comprising SEQ ID NO: 19 and a light chain amino acid sequence comprising SEQ ID NO: 20. Thus, the antibody or antigen binding protein may bind to EGFR and comprise: a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 11 , HCDR2 of SEQ ID NO: 12 and HCDR3 of SEQ ID NO: 13, and wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 14, LCDR2 of SEQ ID NO: 15 and LCDR3 of SEQ ID NO: 16, and wherein the antibody or antigen-binding protein comprises a heavy chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 19, and a light chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20.

[0682] The antibody or antigen-binding protein may bind to HER-2. The antibody or antigen-binding protein may bind to HER-2 and comprise a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 21 , HCDR2 of SEQ ID NO: 22 and HCDR3 of SEQ ID NO: 23, and wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 24, LCDR2 of SEQ ID NO: 25 and LCDR3 of SEQ ID NO: 26.

[0683] The antibody or antigen-binding protein may bind to HER-2 and comprise: a heavy chain CDR1 having 1 , 2 or 3 amino acid differences when compared to the HCDR1 of SEQ ID NO: 21 ; a heavy chain CDR2 having 1 , 2 or 3 amino acid differences when compared to the HCDR2 of SEQ ID NO: 22; a heavy chain CDR3 having 1 , 2 or 3 amino acid differences when compared to the HCDR3 of SEQ ID NO: 23; a light chain CDR1 having 1 , 2 or 3 amino acid differences when compared to the LCDR1 of SEQ ID NO: 24; a light chain CDR2 having 1 , 2 or 3 amino acid differences when compared to the LCDR2 of SEQ ID NO: 25; and / or (e.g., and) a light chain CDR3 having 1 , 2 or 3 amino acid differences when compared to the LCDR3 of SEQ ID NO: 26.

[0684] The antibody or antigen-binding protein may bind to HER-2 and comprise a VH region amino acid sequence comprising SEQ ID NO: 27 and a VL region amino acid sequence comprising SEQ ID NO: 28.

[0685] The antibody or antigen-binding protein may bind to HER-2 and comprise a VH region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 27. The antibody or antigen-binding protein may bind to HER-2 and comprise a VL region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 28. The CDRs of such an antibody or antigen-binding protein may be identical to the CDRs of an antibody or antigen-binding protein comprising a VH region amino acid sequence comprising SEQ ID NO: 27 and a VL region amino acid sequence comprising SEQ ID NO: 28. Thus, the antibody or antigen binding protein may bind to HER-2 and comprise: a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 21 , HCDR2 of SEQ ID NO: 22 and HCDR3 of SEQ ID NO: 23, and wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 24, LCDR2 of SEQ ID NO: 25 and LCDR3 of SEQ ID NO: 26, and wherein the antibody or antigen-binding protein comprises a VH region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 27, and a VL region amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 28.

[0686] The antibody or antigen-binding protein may bind to HER-2 and comprise a heavy chain amino acid sequence comprising SEQ ID NO: 29 and a light chain amino acid sequence comprising SEQ ID NO: 30.

[0687] The antibody or antigen-binding protein may bind to HER-2 and comprise a heavy chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 29. The antibody or antigen-binding protein may bind to HER-2 and comprise a light chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 30. The CDRs of such an antibody or antigen-binding protein may be identical to the CDRs of an antibody or antigen-binding protein comprising a heavy chain amino acid sequence comprising SEQ ID NO: 29 and a light chain amino acid sequence comprising SEQ ID NO: 30. Thus, the antibody or antigen binding protein may bind to HER-2 and comprise: a heavy chain variable (VH) region having a set of CDRs HCDR1 , HCDR2, and HCDR3 and a light chain variable (VL) region having a set of CDRs LCDR1 , LCDR2 and LCDR3, wherein the VH region amino acid sequence comprises HCDR1 of SEQ ID NO: 21 , HCDR2 of SEQ ID NO: 22 and HCDR3 of SEQ ID NO: 23, and wherein the VL region amino acid sequence comprises LCDR1 of SEQ ID NO: 24, LCDR2 of SEQ ID NO: 25 and LCDR3 of SEQ ID NO: 26, and wherein the antibody or antigen-binding protein comprises a heavy chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 29, and a light chain amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 30.

[0688] The antibody or antigen-binding protein may be alemtuzumab, atezolizumab, amivantamab, avelumab, cemiplimab, cetuximab, dratumumab, dinutuximab, durvalumab, elotuzumab, ipilimumab, isatuximab, margetuximab, mogamulizumab, naxitamab, necitumumab, nivolumab, obinutuzumab, ofatumumab, pembrolizumab, pertuzumab, relatlimab, rituximab, tafasitamab, or trastuzumab, or a biosimilar thereof. The antibody or antigen-binding protein may be trastuzumab or a biosimilar thereof.

[0689] Where the antibody or antigen-binding protein is trastuzumab or a biosimilar thereof, the antigen may be HER-2, and / or the target cell may be a breast cell (e.g., a breast cancer cell). For example, where the antibody is trastuzumab or a biosimilar thereof, the antigen may be HER-2 and the target cell may be a breast cell (e.g., a breast cancer cell).

[0690] When the disorder is a CD20+ cancer (such as a CD20+ non-Hodgkin lymphoma, NHL, or CD20+ chronic lymphocytic leukaemia, CLL) the antibody or antigen binding protein may comprise an ADCC-promoting anti-CD20 antibody or antigen-binding protein, such as rituximab or obinutuzumab (or a biosimilar of such an antibody). Thus, ADCC of CD20+ target cancer cells may be induced.

[0691] When the disorder is a CD38+ cancer (such as a CD38+ multiple myeloma) the antibody or antigen binding protein may comprise an ADCC-promoting anti-CD38 antibody or antigenbinding protein, such as daratumumab (or a biosimilar thereof). Thus, ADCC of CD38+ target cancer cells may be induced.

[0692] When the disorder is a SLAMF7+ cancer (such as a SLAMF7+ multiple myeloma) the antibody or antigen binding protein may comprise an ADCC-promoting anti-SLAMF7 antibody or antigen-binding protein, such as elotuzumab (or a biosimilar thereof). Thus, ADCC of SLAMF7+ target cancer cells may be induced.

[0693] When the disorder is a HER2+ cancer (such as a HER2+ breast cancer or HER2+ gastric cancer) the antibody or antigen binding protein may comprise an ADCC-promoting anti-HER2 antibody or antigen-binding protein, such as trastruzumab, pertuzumab, or margetuximab (or a biosimilar of such an antibody). Thus, ADCC of HER2+ target cancer cells may be induced.

[0694] When the disorder is an EGFR+ cancer (such as an EGFR+ head and neck cancer, optionally an EGFR+ head and neck squamous cell carcinoma, or an EGFR+ colorectal cancer) the antibody or antigen binding protein may comprise an ADCC-promoting anti-EGFR antibody or antigen-binding protein, such as cetuximab (or a biosimilar thereof). Thus, ADCC of EGFR+ target cancer cells may be induced.

[0695] When the disorder is a CD19+ cancer (such as a CD19+ multiple myeloma) the antibody or antigen binding protein may comprise an ADCC-promoting anti-CD19 antibody or antigenbinding protein, such as tafasitamab (or a biosimilar thereof). Thus, ADCC of CD19+ target cancer cells may be induced. The antibody or antigen-binding protein may be an antibody. The antibody or antigen-binding protein may be an antigen-binding protein. The antigen-binding protein may be any suitable antigen-binding protein which comprises a domain for binding to an ADCC-promoting FcR on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The antigen-binding protein may comprise a Fab domain, a Fv domain, a scFv domain, a dAb domain, a Fd domain, a Fab’ domain, a F(ab’)2domain or a complementarity determining region (CDR) having sufficient framework to bind an antigen.

[0696] In one embodiment, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not be used in combination with a cell engaging therapy, such as a T cell engaging therapy. In one embodiment, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not be used in combination with an antibody or antigen-binding protein selected from the group consisting of: bispecific T cell engagers (BiTEs); checkpoint-inhibitory T cell engagers (CiTEs); simultaneous multiple interaction T cell engagers (SMiTEs); trispecific killer engagers (TRiKEs); and BiTE-expressing CAR-T cells (CART.BiTE cells). In one embodiment, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not be used in combination with T cell engaging therapies such as mono / bispecific 4-1 BB agonists, or TAA / 4-1 BB bispecific T cell engagers or mono / bispecific 0X40 agonists. In one embodiment, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not be used in combination with mono / bispecific antibodies that activate innate immune cells. In one embodiment, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may not be used in combination with anti-CD40 mAb or anti-CD40 / TAA bispecific. Thus, in one embodiment, the antibody or antigen-binding protein may not be a cell engaging therapeutic, such as a T cell engaging therapeutic. In one embodiment, the antibody or antigen-binding protein may not be selected from the group consisting of: a bispecific T cell engager (BiTE); a checkpoint-inhibitory T cell engager (CiTE); a simultaneous multiple interaction T cell engager (SMiTE); a trispecific killer engager (TRiKE); and a BiTE-expressing CAR-T cell (CART.BiTE cell). In one embodiment, the antibody or antigen-binding protein may not be a T cell engaging therapy such as a mono / bispecific 4-1 BB agonist, or a TAA / 4-1 BB bispecific T cell engager, or a mono / bispecific 0X40 agonist. In one embodiment, the antibody or antigen-binding protein may not be a mono / bispecific antibody that activates innate immune cells. In one embodiment, the antibody or antigen-binding protein may not be an anti-CD40 mAb or anti-CD40 / TAA bispecific.

[0697] In one embodiment, the antibody or antigen-binding protein may not be an antibody selected from the group consisting of: a CD49f antibody, a CD10 antibody, a CD90 antibody, a CD3 antibody, a CD16 antibody, a CD19 antibody, a CD20 antibody, a CD14 antibody, a CD56 antibody, a CD34 antibody, a CD123 antibody, a CD38 antibody, a CD133 antibody, a CD45RA antibody, a CD11 b antibody, a CD71 antibody, a CD66b antibody, a CD49d antibody, a CD117 antibody, a CD16 antibody, a CD62L antibody, a CD54 antibody, a CD63 antibody, a CD18 antibody, a CD15 antibody, a CD11c antibody, a HLA-DR antibody, a CD16 antibody, a CD206 antibody, a CD68 antibody, a CD4 antibody, a CD8 antibody, a CD56 antibody, a CD19 antibody, a CD71 antibody, a CD115 antibody, a CXCR4 antibody, a CD64 antibody, a CD32 antibody, a CXCR2 antibody, a CD40 antibody, a CD62L antibody, a CD18 antibody, a CD14 antibody, a CD206 antibody, a CD68 antibody, a CD4 antibody, a CD8 antibody, a CD56 antibody, a CD107a antibody, a 4-1 BB antibody, and an 0X40 antibody.

[0698] In one embodiment, the antibody or antigen-binding protein may not be an antibody which binds to a marker listed in Tables 7-12. In one embodiment, the antibody or antigen-binding protein may not be an antibody or antigen-binding protein listed in Tables 7-12. In one embodiment, the antibody or antigen-binding protein may not be an antibody selected from the group consisting of: a CD49f antibody, a CD10 antibody, a CD90 antibody, a CD3 antibody, a CD16 antibody, a CD19 antibody, a CD20 antibody, a CD14 antibody, a CD56 antibody, a CD34 antibody, a CD123 antibody, a CD38 antibody, a CD133 antibody, a CD45RA antibody, a CD11 b antibody, a CD71 antibody, a CD66b antibody, a CD49d antibody, a CD117 antibody, a CD16 antibody, a CD62L antibody, a CD54 antibody, a CD63 antibody, a CD18 antibody, a CD15 antibody, a CD11c antibody, a HLA-DR antibody, a CD16 antibody, a CD206 antibody, a CD68 antibody, a CD4 antibody, a CD8 antibody, a CD56 antibody, a CD19 antibody, a CD71 antibody, a CD115 antibody, a CXCR4 antibody, a CD64 antibody, a CD32 antibody, a CXCR2 antibody, a CD40 antibody, a CD62L antibody, a CD18 antibody, a CD14 antibody, a CD206 antibody, and a CD68 antibody.

[0699] In one embodiment, the antibody or antigen-binding protein may not be an antibody selected from the group consisting of: a CD3 antibody, a CD4 antibody, a CD8 antibody, a CD56 antibody, a CD107a antibody, a 4-1 BB antibody, and an 0X40 antibody. In one embodiment, the antibody or antigen-binding protein may not be an antibody selected from the group consisting of: CD3 (OKT3), CD4 (RPA-T4), CD8 (RPA-T8), CD56 (HCD56), CD107a (H4A3), 4-1 BB (4B4-1) and 0X40 (Ber-ACT35)

[0700] The invention relates to medical uses involving a combination of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) that is CD16- and expresses an ADCC-promoting Fc receptor (FcR) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR. The invention also relates to a combination of a composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) with the antibody or antigen-binding protein.

[0701] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) or composition and the antibody or antigen-binding protein may be administered in any suitable order. Thus, the administration may comprise administering the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and the antibody or antigen-binding protein to the subject simultaneously or sequentially. The administration may comprise administering the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) to a subject that has been administered, is being administered, or will be administered the antibody or antigen-binding protein. The administration may comprise administering the antibody or antigen-binding protein to a subject that has been administered, is being administered, or will be administered the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . The administration may comprise administering the composition and the antibody or antigen-binding protein to the subject simultaneously or sequentially. The administration may comprise administering the composition to a subject that has been administered, is being administered, or will be administered the antibody or antigen-binding protein. The administration may comprise administering the antibody or antigen-binding protein to a subject that has been administered, is being administered, or will be administered the composition.

[0702] The administration may comprise administering the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) to the subject as part of a treatment regimen that comprises administering the antibody or antigen-binding protein to the subject. The administration may comprise administering the composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) to the subject as part of a treatment regimen that comprises administering the antibody or antigen-binding protein to the subject. The administration may comprise administering the antibody or antigen-binding protein to the subject as part of a treatment regimen that comprises administering the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) to the subject.

[0703] For the avoidance of doubt, it is noted that the party administering the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) (or composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ) may not necessarily be the same party who has administered, is administering, or will administer another component (e.g., the antibody or antigen-binding protein) of the treatment regimen. Similarly, it is noted that the party administering the antibody or antigen-binding protein may not necessarily be the same party who has administered, is administering, or will administer another component (e.g., the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) or the composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) ) of the treatment regimen. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and the antibody or antigen-binding protein may be administered by different parties. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and the antibody or antigen-binding protein may be administered by the same party. The antibody or antigenbinding protein and the composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be administered by different parties. The antibody or antigenbinding protein and the composition comprising the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may be administered by the same party.

[0704] Administration may be by any suitable technique or route, including but not limited to intravenous injection, intra-arterial injection, intraperitoneal injection, injection into a tumour resection cavity, intrathecal injection, or combinations thereof. Preferably, the administration may be intravenously.

[0705] An appropriate dosage range of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , of a composition of the invention, or of an antibody or antigen-binding protein of the invention is one that produces the desired therapeutic effect when provided to a recipient. An appropriate dosage range may be achieved by a single incidence of administration, or by multiple incidences of administration.

[0706] A typical treatment regimen may include administering from 106, 107, 108or 109cells (e.g. granulocytes or precursors thereof) to a subject, or up to 1012, 1013or 1014cells to a subject. In a suitable embodiment, a treatment regimen includes administering a dose of at least 1 x 109cells to a subject. For example, a treatment regimen may include administering a dose of at least 2 x 109cells or at least 5 x 109cells to a subject. A treatment regimen may include administering a dose of at least 1 x 1010cells or at least 5 x 1010cells to a subject. At least 1 x 1011or at least 2 x 1011cells may be administered to a subject. Between 1 x 109to 3 x 1011or 1 x 1010to 3 x 1011cells may be administered to a subject. For example, between 5 x 1010to 2.5 x 1011cells may be administered to a subject. Meanwhile, the antibody or antigen-binding protein may be administered at any suitable dosage. A subject for treatment may be dosed once, twice, three times, four times, five times, or six times per week. Alternatively, a subject may be dosed daily (e.g. once or twice daily). In other embodiments a subject may be dosed once weekly or bi-weekly. The dose may be weekly. The skilled person will appreciate that the dose can be tailored based on the needs of the subject, and efficacy of the medicament. For example, where the medicament is highly efficacious, the dose may be lowered.

[0707] A subject for treatment may be dosed weekly (e.g. once weekly) with at least 2 x 109cells or at least 2 x 1010cells. A subject for treatment may be dosed weekly with at least 1 x 1011or at least 2 x 1011cells.

[0708] The treatment term can be varied based on the response of the subject to the treatment, and / or the type and / or severity of the disorder (e.g. cancer or infection). For example, the subject for treatment may be dosed for at least 1 or 2 weeks. The subject for treatment may be dosed for at least 3 or 4 weeks. The subject for treatment may be dosed for at least 5 or 6 weeks, suitably at least 7 or 8 weeks.

[0709] A subject for treatment may be dosed for 4-8 weeks with at least 2 x 109cells, wherein said cells are administered once weekly. A subject for treatment may be dosed for 8 weeks with at least 2 x 109cells (e.g. at least 2 x 1010or 2 x 1011cells), wherein said cells may be administered once weekly.

[0710] The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , antibody or antigenbinding protein, and combinations described herein may be for use in treating a disorder by inducing ADCC of the target, e.g., a target cell described herein. Thus, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may treat the disorder by inducing ADCC of the target. The antibody or antigen-binding protein may treat the disorder by inducing ADCC of the target. The granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and antibody or antigen-binding protein may treat the disorder by inducing ADCC of the target.

[0711] As described elsewhere herein, ADCC of targets may be induced by an antibody or antigenbinding protein binding to an Fc receptor on an immune effector cell. Thus, ADCC of the target may be induced by the antibody or antigen-binding protein binding to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase expression of one or more markers of degranulation on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigen-binding protein to the ADCC- promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase expression of one or more costimulatory molecules on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigenbinding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase proliferation of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase abundance of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigen-binding protein to the ADCC- promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase survival of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase expression of one or more cytokines by the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Binding of the antibody or antigen-binding protein to the ADCC- promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase trafficking of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . Preferably, binding of the antibody or antigen-binding protein to the ADCC- promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase cytocidal activity of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0712] Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase some or all of the properties of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) set out above.

[0713] ADCC of targets may be induced by an antibody or antigen-binding protein binding to an Fc receptor on an immune effector cell and activating the immune effector cell. Thus, ADCC of the target may be induced by the antibody or antigen-binding protein binding to the ADCC- promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and activating the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) . ADCC of the target may be induced by the antibody or antigen-binding protein binding to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and increasing activation of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) .

[0714] Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that expression by the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) of one or more markers of degranulation is increased. Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that expression by the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) of one or more costimulatory molecules is increased. Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that proliferation of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is increased. Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that abundance of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is increased. Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that survival of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is increased. Binding of the antibody or antigenbinding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that expression by the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) of one or more cytokines is increased. Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that trafficking of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is increased. Preferably, binding of the antibody or antigen-binding protein to the ADCC- promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may activate, or increase activation of, the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) such that cytocidal activity of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is increased.

[0715] Binding of the antibody or antigen-binding protein to the ADCC-promoting Fc receptor on the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) may increase activation of the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) by some or all of the properties set out above.

[0716] The disorder may be any suitable disorder. The disorder may be any disorder which is susceptible to treatment by ADCC, e.g., by ADCC of disordered cells. The disorder may be any disorder which is susceptible to treatment with an antibody or antigen-binding protein comprising a domain for binding to an ADCC-promoting FcR. The disorder may be any disorder which is susceptible to treatment with an Fc-enabled antibody or antigen-binding protein.

[0717] As used herein, reference to a “disorder” includes reference to a disease. Thus, the disorder may be a disease. The disease may be any suitable disease. The disease may be any disease which is susceptible to treatment by ADCC, e.g., by ADCC of diseased cells. The disease may be any disease which is susceptible to treatment with an antibody or antigen-binding protein comprising a domain for binding to an ADCC-promoting FcR. The disease may be any disease which is susceptible to treatment with an Fc-enabled antibody or antigen-binding protein. For example, the disease may be cancer. Where the disease is cancer, the target may be a cancer cell. The cancer may be a solid tumour or a liquid tumour. Preferably, the cancer may be a solid tumour.

[0718] The term “solid tumour” may refer to an abnormal, malignant mass of tissue that does not contain cysts or liquid inclusions. Examples of solid tumour cancers include carcinomas, sarcomas, and lymphomas.

[0719] A solid tumour cancer may be a carcinoma. A carcinoma may be selected from one or more of an adenocarcinoma, a basal cell carcinoma, a squamous cell carcinoma, an adenosquamous carcinoma, a renal cell carcinoma, a ductal carcinoma in situ (DCIS), an invasive ductal carcinoma, an anaplastic carcinoma, a large cell carcinoma, a small cell carcinoma or combinations thereof. A carcinoma may also be selected from epithelial neoplasms, squamous cell neoplasms, squamous cell carcinoma, basal cell neoplasms, basal cell carcinoma, transitional cell carcinomas, adenocarcinomas (such as Adenocarcinoma not otherwise specified (NOS), linitis plastica, vipoma, cholangiocarcinoma, hepatocellular carcinoma NOS, adenoid cystic carcinoma, renal cell carcinoma, Grawitz tumour), adnexal and skin appendage neoplasms, mucoepidermoid neoplasms, cystic mucinous and serous neoplasms, ductal lobular and medullary neoplasms, acinar cell neoplasms, or complex epithelial neoplasms. A solid tumour cancer may be a squamous cell carcinoma of the head or neck.

[0720] Alternatively, a solid tumour cancer may be a sarcoma. A sarcoma may be selected from Askin's tumour, sarcoma botryoides, chondrosarcoma, Ewing's, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, or soft tissue sarcomas (including alveolar soft part sarcoma, angiosarcoma, cystosarcoma phyllodes, dermatofibrosarcoma protuberans (DFSP), desmoid tumour, desmoplastic small round cell tumour, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumour (GIST), hemangiopericytoma, hemangiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, malignant fibrous histiocytoma, undifferentiated pleomorphic sarcoma, malignant peripheral nerve sheath tumour (MPNST), neurofibrosarcoma, rhabdomyosarcoma, and synovial sarcoma).

[0721] Alternatively, a solid tumour may be a lymphoma, such as a B-cell lymphoma, a T-cell lymphoma, a NK-cell lymphoma, or a Hodgkin’s lymphoma.

[0722] The disease may be cancer and the antibody or antigen-binding protein may bind to a cancer antigen. The antibody or antigen-binding protein may specifically bind to a cancer antigen. The cancer antigen may be HER-2.

[0723] As set out in Example 18, the inventors have found that the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention are able to bring about ADCC-mediated killing of cancer cells derived from head and neck cancer (in particular, killing cancer cells from a head and neck squamous cell carcinoma - HNSCC). This is of particular clinical value in that such cancers may otherwise be difficult to treat via other therapeutic modalities. Patients with EGFR+ cancers (such as the HNSCC forming the basis of the model used in Example 18) often experience low levels of response, or relapse, to conventional anti-EGFR therapies. The compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may help to overcome this failing.

[0724] Thus, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in the treatment of cancers that have proven resistant to treatment, that have exhibited low response to treatment, or that have undergone relapse. The compositions, granulocyte precursor cells, and antibodies (or antigenbinding proteins) for use in accordance with the invention may be used as a second line therapy in the treatment of a cancer that has proven resistant to treatment by a previous therapy, that has exhibited low response to treatment by a previous therapy, or that has undergone relapse after treatment with a previous therapy.

[0725] In a suitable embodiment, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in combination with a further therapeutic agent or regimen in the treatment of a cancer. In a suitable embodiment, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in combination with an immunotherapy, with chemotherapy, or with radiotherapy the treatment of a cancer.

[0726] In a suitable embodiment, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in the treatment of a head and neck cancer. In a suitable embodiment, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in the treatment of an HNSCC cancer.

[0727] In a suitable embodiment, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in the treatment of an EGFR+ cancer. In such cases, the antibody (or antigen-binding protein) suitably comprises an anti-EGFR antibody, and the target is an EGFR+ cancer cell. In a suitable embodiment, the compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used in the treatment of an EGFR+ that has previously received treatment with another immunotherapy, with chemotherapy, or with radiotherapy. The compositions, granulocyte precursor cells, and antibodies (or antigenbinding proteins) for use in accordance with the invention may be used as a second line therapy in the treatment of an EGFR+ cancer that has proven resistant to treatment by a previous therapy, that has exhibited low response to treatment by a previous therapy, or that has undergone relapse after treatment with a previous therapy. The compositions, granulocyte precursor cells, and antibodies (or antigen-binding proteins) for use in accordance with the invention may be used as a second line therapy in the treatment of a cancer that has proven resistant to treatment by another previous anti-EGFR immunotherapy therapy, that has exhibited low response to treatment by another previous anti-EGFR immunotherapy, or that has undergone relapse after treatment with another previous anti-EGFR immunotherapy.

[0728] The disease may be an infection. Where the disease is an infection, the target may be a pathogen, such as a pathogen cell. Where the disease is an infection, the antigen may be an antigen expressed by a pathogen, e.g., an antigen which is expressed by the pathogen but not by a cell of the subject.

[0729] The infection may be a bacterial infection, a fungal infection, a viral infection, a macroparasite infection (e.g., a helminth infection), or a combination thereof. The infection may be caused by a pathogen. The infection may be caused by a bacterium. The infection may be caused by a Gram-negative bacterium or a Gram-positive bacterium. Preferably, the infection may be caused by a Gram-positive bacterium, such as a bacterium from the genus Staphylococcus.

[0730] The infection may be caused by a bacterium selected from one or more of Staphylococcus spp., multidrug resistant gram-negative bacteria (MRDGN bacteria), vancomycin-resistant Enterococcus (VRE), Mycobacterium spp., carbapenem-resistant Enterobacteriaceae (CRE) gut bacteria, Acinetobacter spp., Actinomyces spp., Propionibacterium spp., Anaplasma spp., Bacillus spp., Arcanobacterium spp., Bacteroides spp., Bartonella spp., Brucella spp., Yersinia spp., Burkholderia spp., Campylobacter spp., Streptococcus spp., Haemophilus spp., Clostridium spp., Corynebacterium spp., Echinococcus spp., Ehrlichia spp., Enterococcus spp., Rickettsia spp., Fusobacterium spp., Neisseria spp., Klebsiella spp., Helicobacter spp., Escherichia spp., Kingella spp., Legionella spp., Listeria spp., Borrelia spp., Mycoplasma spp., Chlamydia spp., Nocardia spp., Pasteurella spp., Bordetella spp., Prevotella spp., Chlamydophila spp., Coxiella spp., Salmonella spp., Group A Streptococcus spp., Shigella spp., Staphylococcus spp., Treponema spp., Vibrio spp., Francisella spp., Pseudomonas spp. and Ureaplasma spp.

[0731] The bacterium may be selected from one or more of methicillin resistant Staphylococcus aureus (MRSA), multi-drug resistant Mycobacterium tuberculosis (MDR-TB), Pseudomonas aeruginosa, Pseudomonas oryzihabitans, Pseudomonas plecoglossicida, Acinetobacter baumannii, Actinomyces israelii, Actinomyces gerencseriae, Propionibacterium propionicus, Bacillus anthracis, Arcanobacterium haemolyticum, Bacillus cereus, Yersinia pestis, Mycobacterium ulcerans, Campylobacter jejuni, Bartonella bacilliformis, Bartonella henselae, Haemophilus ducreyi, Clostridium difficile, Corynebacterium diphtheria, Burkholderia mallei, Neisseria gonorrhoeae, Klebsiella granulomatis, Streptococcus pyogenes, Streptococcus agalactiae, Haemophilus influenzae, Helicobacter pylori, Escherichia coll (e.g. O157:H7, 0111 and O104:H4), Kingella kingae, Legionella pneumophila, Listeria monocytogenes, Burkholderia pseudomallei, Neisseria meningitidis, Mycoplasma pneumoniae, Mycoplasma genitalium, Chlamydia trachomatis, Bordetella pertussis, Streptococcus pneumoniae, Chlamydophila psittaci, Coxiella burnetii, Treponema pallidum, Clostridium tetani, Chlamydophila pneumoniae, Vibrio cholera, Mycobacterium tuberculosis, Salmonella enterica subsp. enterica, serovartyphi, Ureaplasma urealyticum, and Francisella tularensis. Preferably Mycobacterium tuberculosis.

[0732] The bacterium may be selected from one or more of methicillin resistant Staphylococcus aureus (MRSA), multidrug resistant gram-negative bacteria (MRDGN bacteria), vancomycin- resistant Enterococcus (VRE), multi-drug resistant Mycobacterium tuberculosis (MDR-TB), and carbapenem-resistant Enterobacteriaceae (CRE) gut bacteria.

[0733] The infection may be caused by a virus selected from one or more family selected from Adenoviridae, Picornaviridae, Herpesviridae, Coronaviridae, Hepadnaviridae, Flaviviridae, Retroviridae, Orthomyxoviridae, Paramyxoviridae, Papovaviridae, Polyomavirus, Rhabdoviridae, Togaviridae and Bunyaviridae.

[0734] The virus may be selected from one or more of HIV-1 (Human immunodeficiency virus), HIV- 2, Junin virus, BK virus, Machupo virus, Sabia virus, Varicella zoster virus (VZV), Alphavirus, Colorado tick fever virus (CTFV), Rhinoviruses, Crimean-Congo hemorrhagic fever virus, Cytomegalovirus, Dengue virus, Ebolavirus (EBOV), Parvovirus B19, Human herpesvirus 6 (HHV-6), Human herpesvirus 7 (HHV-7), Enteroviruses (e.g. EV71), Coxsackie A virus, Sin Nombre virus, Heartland virus, Hanta virus, Hendra virus, Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Hepatitis D Virus, Hepatitis E virus, Herpes simplex virus 1 and 2 (HSV-1 and HSV-2), Human bocavirus (HBoV), Human metapneumovirus (hMPV), Human papillomaviruses, Human parainfluenza viruses (HPIV), Epstein-Barr virus (EBV), Lassa virus, Lymphocytic choriomeningitis virus (LCMV), Marburg virus, Measles virus, Middle East respiratory syndrome coronavirus, Molluscum contagiosum virus (MCV), Monkeypox virus, Mumps virus, Nipah virus, Norovirus, Poliovirus, JC virus, Respiratory syncytial virus (RSV), Rhinovirus, Rift Valley fever virus, Rotavirus, Rubella virus, SARS coronavirus, SARS-CoV-2, Variola major, Variola minor, Venezuelan equine encephalitis virus, Guanarito virus, West Nile virus, Yellow fever virus, and Zika virus.

[0735] The infection may be caused by a fungus. The infection may be caused by a fungus selected from one or more of Aspergillus spp., Piedraia spp., Blastomyces spp., Candida spp., Fonsecaea spp., Coccidioides spp., Cryptococcus spp., Cryptosporidium spp., Geotrichum spp., Histoplasma spp., Microsporidia phylum, Paracoccidioides spp., Pneumocystis spp., Sporothrix spp., Trichophyton spp., Epidermophyton spp., Hortaea spp., Malassezia spp., Trichosporon spp., and Mucorales order.

[0736] The fungus may be selected from one or more of Aspergillus fumigatus, Aspergillus flavus, Piedraia hortae, Blastomyces dermatitidis, Candida albicans, Fonsecaea pedrosoi, Coccidioides immitis, Coccidioides posadasii, Cryptococcus neoformans, Geotrichum candidum, Histoplasma capsulatum, Paracoccidioides brasiliensis, Pneumocystis jirovecii, Sporothrix schenckii, Trichophyton tonsurans, Epidermophyton floccosum, Hortaea werneckii, and Trichosporon beigelii.

[0737] The infection may be caused by a microparasite. The macroparasite may be one or more selected from Angiostrongylus spp., Entamoeba Anisakis spp., Ascaris spp., Babesia spp., Balantidium spp., Baylisascaris spp., Blastocystis spp., Capillaria spp., Trypanosoma spp., Clonorchis spp., Ancylostoma spp., Cyclospora spp., Taenia spp., Desmodesmus spp., Dientamoeba spp., Dracunculus spp,. Enterobius spp., Fasciola spp., Filarioidea superfamily, Giardia spp., Gnathostoma spp., Necator spp., Hymenolepis spp., Isospora spp., Leptospira spp., Wuchereria spp., Rhinosporidium spp., Brugia spp., Plasmodium spp., Onchocerca spp., Opisthorchis spp., Paragonimus spp., Naegleria spp., Schistosoma spp., Strongyloides spp., Toxocara spp., Toxoplasma spp., Trichinella spp., Trichomonas spp., and Trichuris spp.

[0738] The macroparasite may be selected from one or more of Entamoeba histolytica, Ascaris lumbricoides, Balanti...

Claims

CLAIMS1. A composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) for use in treating a disorder in a subject in combination with an antibody or antigen-binding protein for binding to an antigen on a target, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc- receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

2. An antibody or antigen-binding protein for binding to an antigen on a target for use in treating a disorder in a subject in combination with a composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) , wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc- receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

3. A composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target for use in treating a disorder in a subject, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc- receptor (FcR), and wherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

4. A composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc- receptor (FcR), andwherein the antibody or antigen-binding protein comprises a domain for binding to the ADCC-promoting FcR.

5. A composition according to claim 4 for use in treating a disorder in a subject.

6. The composition for use, or the antibody or antigen-binding protein for use according to any one of the preceding claims, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and antibody or antigen-binding protein treat the disorder by inducing ADCC of the target.

7. The composition for use, the antibody or antigen-binding protein for use, or the composition according to any one of the preceding claims, wherein the ADCC-promoting FcR is an ADCC-promoting IgG or IgA FcR.

8. The composition for use, the antibody or antigen-binding protein for use, or the composition according to claim 7, wherein the ADCC-promoting FcR is CD64, CD32, CD89 or a combination thereof.

9. The composition for use, the antibody or antigen-binding protein for use, or the composition according to any one of the preceding claims, wherein the domain for binding to the ADCC-promoting FcR is an Fc domain, optionally an IgG or IgA Fc domain.

10. The composition for use, the antibody or antigen-binding protein for use, or the composition according to any one of the preceding claims, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) expresses neutrophil elastase.

11. The composition for use, the antibody or antigen-binding protein for use, or the composition according to any one of the preceding claims, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) expresses CD64 and neutrophil elastase.

12. The composition for use, the antibody or antigen-binding protein for use, or the composition according to any one of the preceding claims, wherein the granulocyte precursor cell has been differentiated in vitro and is CD16- and comprises: i. increased expression of one or more of: SRGN, MPO, HLA-DRA, CD74, and ELANE when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo,and / or ii. decreased expression of one or more of: DEFA1, DEFA3, CAMP, BPI, and AZU1 when compared to an equivalent granulocyte precursor cell that has been differentiated in vivo.

13. The composition for use, the antibody or antigen-binding protein for use, or the composition according to any one of the preceding claims, wherein the CD16- granulocyte or precursor thereof (e.g., a granulocyte precursor cell) has been derived from a donor having a granulocyte (neutrophil) with high CKA.

14. The composition for use, or the antibody or antigen-binding protein for use, or the composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for use according to any one of claims 1-3 or 5-13, wherein the disorder is cancer and the target is a cancer cell.

15. The composition for use, or the antibody or antigen-binding protein for use, or the composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for use according to claim 14, wherein the cancer is selected from: head and neck cancer, cervical cancer (such as metastatic cervical cancer), pancreatic cancer, liver cancer, oesophageal cancer, stomach cancer, ovarian cancer, lung cancer, bladder cancer, kidney cancer, brain cancer, prostate cancer, myeloma cancer, nonHodgkin’s lymphoma (NHL), larynx cancer, uterine cancer, or breast cancer.

16. The composition for use, or the antibody or antigen-binding protein for use, or the composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for use according to claim 14 or claim 15, wherein the antibody (or antigen binding protein) binds an antigen associated with a head and neck cancer cell, a cervical cancer cell (such as metastatic cervical cancer cell), a pancreatic cancer cell, a liver cancer cell, an oesophageal cancer cell, a stomach cancer cell, an ovarian cancer cell, a lung cancer cell, a bladder cancer cell, a kidney cancer cell, a brain cancer cell, a prostate cancer cell, a myeloma cancer cell, a non-Hodgkin’s lymphoma (NHL) cell, a larynx cancer cell, a uterine cancer cell, or a breast cancer cell.

17. The composition for use, or the antibody or antigen-binding protein for use, or the composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for use according to any of claims 14 to 16, wherein the antibody or antigen-binding protein binds to HER-2.

18. The composition for use, or the antibody or antigen-binding protein for use, or the composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for use according to any of claims 14 to 17, wherein the antibody or antigen-binding protein binds to EGFR.

19. The composition for use, or the antibody or antigen-binding protein for use according to any one of claims 1-3 or 5-13, wherein the disorder is an infection and the target is a pathogen.

20. A method for treating a disorder, the method comprising administering to a subject in need thereof a therapeutically effective amount of a composition or antibody or antigen-binding protein according to any of claims 1 to 19.21 . Use of a composition or antibody or antigen-binding protein according to any of claims 14 to 18, in the manufacture of a medicament for treating cancer.

22. A method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a composition or antibody or antigen-binding protein according to any of claims 14 to 18.

23. Use of a composition or antibody or antigen-binding protein according to any of claim 19, in the manufacture of a medicament for treating an infection.

24. A method for treating an infection, the method comprising administering to a subject in need thereof a therapeutically effective amount of a composition or antibody or antigen-binding protein according to claim 19.

25. A kit comprising the composition according to any one of claims 4 or 7-13.

26. A method for manufacturing a composition or a kit, wherein the method comprises combining a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc-receptor (FcR), and wherein the antibody, or antigen-binding protein, comprises a domain for binding to the ADCC-promoting FcR.

27. Use of a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for binding to an antigen on a target in a method of manufacturing a composition or a kit, wherein the target is a diseased cell or a pathogen, wherein the granulocyte or precursor thereof (e.g., a granulocyte precursor cell) is CD16- and expresses an antibody-dependent cellular cytotoxicity (ADCC)-promoting Fc- receptor (FcR), and wherein the antibody, or antigen-binding protein, comprises a domain for binding to the ADCC-promoting FcR.

28. A composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) for use as described herein.

29. A composition comprising an antibody or antigen-binding for use as described herein.

30. A composition comprising a granulocyte or precursor thereof (e.g., a granulocyte precursor cell) and an antibody or antigen-binding protein for use as described herein.

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