Innate Immunity Cell Identity Markers

Innate Immunity

Rapid responses to invading pathogens are mediated by the cells and humoral factors of the innate immune system. These responses are activated by the interaction between pathogen antigens and pattern recognition receptors (PRRs), localized to the surface or intracellular compartments of innate immune cells. This initial response is somewhat generic, as receptors are only able to recognize a limited repertoire of motifs from pathogen-derived molecules including lipopolysaccharides, lipoproteins and nucleic acids, collectively referred to as pathogen-associated molecular patterns (PAMPs).

Complement C3 expression in mouse kidney cells IHC

TLR3 expression in mouse colon IHC

Immunohistochemistry: Complement C3 Antibody (11H9) [NB200-540] - C3 protein fragments deposited on kidney cells of MPL-lpr mouse. Staining with antibody 11H9. Glomerular staining pattern. Fixation in 4% paraformaldehyde in PBS pH 7.4. Vibratome sections of 4 um. Pretreated with 3% hydrogen peroxide for 20 min to quench endogenous peroxidases. Microwaved in antigen unmasking solution for 2-5 minutes as antigen retrieval.

Immunohistochemistry-Paraffin: TLR3 Antibody (40C1285.6) [NBP2-24875] - Mouse colon using TLR3 antibody (clone 40C1285.6) at 1:500 dilution with HRP-DAB detection and hematoxylin counterstaining. Intense signal was found in subset of cells at the bases of the crypts.



In addition to PAMPs, innate immune cells respond to molecular patterns associated with tissue injury or damage-associated molecular patterns (DAMPs). These are cell-derived molecules induced by stress or injury which may be released even in the absence of infection. Both PAMPs and DAMPs engage innate cellular mediators, including cytotoxic and phagocytic cells via interaction with Toll-like (TLRs), NOD-like, RIG-I-like and AIM2-like receptors. Soluble mediators of the innate response include cytokines, complement and acute phase proteins.


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Innate cellular mediators: functions and markers


Cell Type Function Markers

Dendritic cells

Dendritic cell

Antigen presenting cells, reside in peripheral tissues where they sequester and process antigens for presentation to lymphocytes. Dendritic cells may be classified into three subsets:Myeloid- CD11c, CD13, CD33, CD11b; Plasmacytoid- CD123, CD303, and CD304; Monocyte-related- CD14, all of which have high expression of MHCII (HLA-DR). 

CD11c, CLIP-170, Fascin, MADDAM, NLDC-145, MHCII (HLA-DR), CD33, CD123, CD16, CD128b

Activation: CD83, CD80, CD86, CD40, MHCII (HLA-DR)

Macrophages (and Monocytes)

Macrophages

Phagocytic cells, participate in both innate and adaptive immunity, in the capture of pathogens and as antigen presenting cells. Macrophages persist and monitor tissue for prolonged periods where they demonstrate distinct tissue-specific phenotypes and origins. While some macrophages are yolk sac-derived, others differentiate from bone marrow-derived monocytes. Based on recent nomenclature, macrophages may be classified into two main subsets: M1 classically activated and M2 alternatively activated macrophages, although more subpopulations continue to be identified. 

CD68, CD64, CSF-1R, F4-80 (mice), CD11b

Activation: M1 classically activated- CD68, CD80, CD86, CD38, CD11c

M2 alternatively activated- CD68, CD163, MMR/CD206, MGL1 , MGL2, Egr2

Monocytes: CD14, CD16, CD115 (mice), Ly-6C (mice)

Mast cells

Mast Cells

Mature mast cells are found in association with a variety of tissues including those with high environmental influence (e.g., mucosal and skin tissues). Mast cells degranulate following stimulation through IgE-dependent and independent mechanisms and release several products including proteases, TNF-alpha, GM-CSF and cytokines.

CD34 (mice), Chymase, CD117/c-kit, Fc epsilon RI

Activation: LAMP-I (CD1-7a), LAMP-2 (CD107b)

Natural killer cells (NK)

Natural killer cells

NK cells recognize infected or transformed cells and target them for lysis via the release of perforins and granzymes. Activated NK cells secrete various cytokines including IL-1, IL-2 and INF-gamma. Two main subsets are identifiable for their differential expression of CD16 and CD56.

CD56, CD16, CD57, CD161, NKp46, NKp30, CD25

Activation: CD69, CD107a, NKp46, NKp30, CD95

Granulocytes

Neutrophils Neutrophils

Basophils Basophils

Eosinophils Eosinophils

Phagocytic cells with short life span (days) characterized by the presence of cytosolic granules containing a variety of immunologically active substances which are microbicidal, cytotoxic, or inflammatory.

Neutrophils: CD16, CD11b, CD16b, CD66c, CD66b, Ly-6G/Gr-1 (mice)

Basophils: CD49b (mice), CD41 (mice), Fc epsilon RI, CD203c, CD123 , CCR3/CD193

Eosinophils: CD11b, CCR3/CD193, F4/80 (mice), Siglec-F (mice), EMR1, Siglec-8

Activation:

Neutrophil: CD11b, CD88, CD18, L-selectin (CD62L), CD32, CD54, CD66b

Basophil: CD41 (mice), CD200R, CD63, CD203c, CD69, CD13, CD164, CD107a, CRTH2/DP2

Eosinophils: CD69, CD63, CD44



Learn about NK cells in anti-cancer therapy





Flow cytometry analysis of B7-2/CD86 expression in Human Ramos lymphoma cell line

Human Ramos lymphoma cell line was stained with: A. Mouse Anti-Human B7-2/CD86 Monoclonal Antibody (Catalog # MAB141, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by PE-conjugated Goat Anti-Mouse IgG secondary antibody (Catalog # F0102B); B. B7-2/CD86 knockout Ramos human lymphoma cell line was stained with Mouse Anti-Human B7-2/CD86 Monoclonal Antibody (Catalog #MAB141, filled histogram) or isotype control antibody (Catalog #MAB002, open histogram) followed by PE-conjugated Goat anti-Mouse IgG Secondary Antibody (Catalog #F0102B). No staining in the B7-2/CD86 knockout Ramos cell line was observed. View our protocol for Staining Membrane-associated Proteins.



CD11b expression in mouse spleen IHCImmunocytochemistry/Immunofluorescence: CD11b Antibody (M1/70.15) [NB600-1327] - ICC/IF analysis of RAW264.7 cells using CD11b antibody (clone M1/70.15) with Dylight 488 conjugated secondary (green). Nuclei and tubulin were stained with DAPI (blue) and Dylight 550 (red).



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