US-12618837-B2 - Method of identifying pro-inflammatory dendritic cells

US12618837B2US 12618837 B2US12618837 B2US 12618837B2US-12618837-B2

Abstract

There is provided a method of identifying pro-inflammatory dendritic cells, the method comprising: determining an expression of CD5, CD14 and/or CD163 in cells, wherein CD5 − , CD14 + and/or GD163 + cells are identified as pro-inflammatory dendritic cells. Also disclosed is a method of characterising inflammation and/or inflammatory disease in a subject, the method comprising: determining a proportion of CD5 − , CD14 + and/or GD163 + dendritic cells in the subject's sample, wherein the proportion positively correlates with the level of inflammation and/or the severity of inflammatory disease in the subject.

Inventors

Florent Ginhoux
Charles Antoine Dutertre

Assignees

AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH

Dates

Publication Date: 20260505
Application Date: 20200626
Priority Date: 20190626

Claims (7)

1 . A method of characterising CD1c + dendritic cells, the method comprising: determining an expression of one or more of CD5, CD14 and CD163 in the dendritic cells, wherein where the dendritic cells are determined to be CD5−, CD14+ and CD163+, identifying the dendritic cells as pro-inflammatory dendritic cells.
2 . The method according to claim 1 , wherein where the dendritic cells are determined to be CD163+CD14+, identifying the dendritic cells as highly pro-inflammatory dendritic cells that are more pro-inflammatory than CD163− or CD14− dendritic cells.
3 . The method according to claim 1 , the method further comprising determining a ratio of CD163+CD14+ dendritic cells to a total number of dendritic cells in a sample.
4 . The method according to claim 1 , the method further comprising determining an expression of one or more of CD11b, CD36, CD64, CD87, CD107a, CD206, CD274, CD354, FcεRIα, HLA-DQ, CD2, CD59, CD81, CD166, CD229, CD271 and Integrin β7 in the dendritic cells.
5 . The method according to claim 1 , wherein the dendritic cells have one or more of the following properties: (i) is a conventional CD1c + dendritic cell 2 (cDC2); (ii) is dependent on IRF4 for differentiation; (iii) is dependent on KLF4 for differentiation; (iv) is dependent on FLT3 ligand (FLT3L) for differentiation; and (v) is capable of activating and/or polarizing T cells.
6 . The method according to claim 1 , the method further comprising determining a ratio of CD163+CD14+ dendritic cells to a total number of dendritic cells having one or more of the properties selected from the group consisting of: (i) a conventional dendritic cell; (ii) dependent on IRF4 for differentiation; (iii) dependent on KLF4 for differentiation; (iv) dependent on FLT3 ligand (FLT3L) for differentiation; and (v) capable of activating and/or polarizing T cells.
7 . A kit for characterising CD1c + dendritic cells, inflammation and/or inflammatory disease, the kit comprising reagents for detecting CD5, CD14 and CD163, wherein the kit further comprises methods for identifying dendritic cells to be pro-inflammatory dendritic cells where the dendritic cells are determined to be CD5−, CD14+ and CD163+.

Description

RELATED APPLICATIONS This application is the U.S. National Stage of International Application No. PCT/SG2020/050369, filed Jun. 26, 2020, which designates the U.S., published in English, and claims priority under 35 U.S.C. § 119 or 365(c) to Singapore Application No. 10201905956V, filed Jun. 26, 2019. The entire teachings of the above applications are incorporated herein by reference. TECHNICAL FIELD The present disclosure relates broadly to a method of characterising mononuclear phagocyte populations, such as dendritic cells, and related kits. BACKGROUND Mononuclear phagocytes (MNP) comprise a heterogeneous population of cells historically assigned to subsets based on their phenotype, ontogeny, transcriptomic profiles and their specialized functions. MNP comprise dendritic cell (DC) and monocyte subsets in the blood and tissues, as well as macrophages and monocyte-derived cells (MC) in tissues. It is important to clearly delineate these subsets to study their functions, as particular MNP subsets are increasingly recognized as having important roles in diseases. A clear method to identify these various subsets in the blood and tissues, as well as in health and disease, is challenging as their phenotypes are often overlapping. DC and MC represent two complementary and integrated functional systems in time and space. DC specifically depend on FMS-like tyrosine kinase 3 (FLT3) for their differentiation and proliferation. They classically include plasmacytoid DC (pDC) that are highly specialized in type I interferon production, and conventional DC (cDC) that are major subsets involved in antigen presentation and modulation of immunity. Importantly, the DC nature of pDC is under debate, as they have been shown to differentiate from B-cell progenitors, and thus could be more related to the innate lymphoid cell family. cDC are further subdivided into two major lineages: cDC1, with superior antigen cross-presentation to CD8+ T cells, and cDC2 that perform a wide spectrum of functions, including antigen presentation to CD4+ T cells and priming to T helper 2 (Th2) and Th17-type responses. Importantly, DC subsets depend on critical transcription factors for their differentiation including IRF8 and BATF3 for cDC1 or IRF4 and KLF4 for cDC2. While cDC1 express several highly specific markers (CLEC9A, XCR1, CADM1) allowing their precise delineation, cDC2 express less specific defining markers that often overlap with monocytes. Monocytes are highly heterogenous, plastic cells that patrol blood vessels and can migrate to tissues, where together with resident macrophages, they have central roles in tissue homeostasis maintenance and inflammation. Monocytes differentiate into MC with features characteristic of both macrophages and DC, the latter mostly during inflammation. MC can acquire a multitude of functional capabilities that are largely determined by the inflammatory milieu to which they are recruited. In human blood, monocyte subsets are defined based on their relative expression of CD14 and CD16—two membrane proteins with supposed restricted expression to monocytes among all circulating MNP. Such restricted expression is now under debate, as in the blood, classical monocytes (cMo; CD14hiCD16−) and CD1c+cDC2 are phenotypically related and form a continuum, with cells falling in between that express markers of both cell types including the cDC2 marker CD1c, and the monocyte markers CD14 and CD11b. Similar intermediate pro-inflammatory cells also accumulate in inflamed tissues and while they seem to be derived from monocytes, they are functionally different due to their cDC2-specialized capacity to stimulate autologous CD4+ T cells. Thus, understanding the currently debated nature of these cells could allow their manipulation in pathologic settings. Thus, there is a need to provide an alternative method of characterising mononuclear phagocyte populations, such as dendritic cells, and related kits. SUMMARY In one aspect, there is provided a method of characterising dendritic cells, the method comprising: determining an expression of one or more of CD5, CD14 and CD163 in the dendritic cells. In one embodiment, where the dendritic cells are determined to be CD5−, CD14+ and/or CD163+, the method comprises identifying the dendritic cells as pro-inflammatory dendritic cells. In one embodiment, wherein where the dendritic cells are determined to be CD163+CD14+, the method comprises identifying the dendritic cells as highly pro-inflammatory dendritic cells that are more pro-inflammatory than CD163− or CD14− dendritic cells. In one embodiment, the method further comprises determining a proportion of CD163+CD14+ dendritic cells. In one embodiment, the method further comprises determining an expression of one or more of CD11b, CD36, CD64, CD87, CD107a, CD206, CD274, CD354, FcεRIα, HLA-DQ, CD2, CD59, CD81, CD166, CD229, CD271 and Integrin β7 in the dendritic cells. In one embodiment, the dendritic cells have one or more of the follo