Table 3.
Phenotyping of CD14 − CD16 ++ , CD14 + CD16 + and CD14 + CD16 low/- cell populations
| Cell marker | CD14 − CD16 ++ average MFI (SE) | CD14 + CD16 + average MFI (SE) | CD14 + CD16 low/- average MFI (SE) | |
|---|---|---|---|---|
| CD1b | 129 (28)a | 113 (12) | 54 (7)b | Antigen presentation Co-stimulatory molecules |
| CD80 | 2349 (378) | 1590 (119) | 1889 (66) | Antigen presentation Co-stimulatory molecules |
| CD86 | 1226 (172)a | 2941 (291) | 2403 (403)b | Antigen presentation Co-stimulatory molecules |
| CD40 | 9166 (1548) | 7896 (1224) | 6839 (967) | Antigen presentation Co-stimulatory molecules |
| MHC DR | 46 014 (12 130) | 63 800 (13650) | 32 720 (9057) | Antigen presentation Co-stimulatory molecules |
| CD11b | 20 755 (4334)a | 32 631 (5299) | 47 893 (9406)b | Antigen presentation Co-stimulatory molecules |
| CD11c | 7575 (3295) | 12 828 (4748) | 21 093 (11339) | Antigen presentation Co-stimulatory molecules |
| CD3 | <10a | 43 (11)b | 36 (8)b | Lymphoid cell markers |
| CD4 | <10 | <10 | <10 | Lymphoid cell markers |
| CD8α | <10 | 28 (3) | 14 (4) | Lymphoid cell markers |
| CD8β | <10 | 10 (5) | <10 | Lymphoid cell markers |
| CD21 | 618 (140)A | 247 (23)B | 136 (18)B | Lymphoid cell markers |
| NKp46 | 38 (16) | 34 (3) | 17 (2) | Lymphoid cell markers |
| CD26 | 147 (37) | 151 (22) | 110 (21) | Lymphoid cell markers |
| CD172a | 96 077 (31018) | 100 076 (31620) | 89 046 (28726) | Myeloid cell markers |
| CD206 | 649 (260) | 1533 (389)a | 191 (32)b | Myeloid cell markers |
| TLR2 | 92 (25) | 299 (41) | 261 (96) | Myeloid cell markers |
| CD163 | 409 (68)A | 1273 (58)B | 1898 (222)C | Myeloid cell markers |
| CD16 | 18 344 (1448)A | 4039 (607)B | 366 (99)C | Myeloid cell markers |
| CD14 | 20 (16)A | 978 (78)B | 1748 (337)B | Myeloid cell markers |
Three colour flow cytometry was carried out on PBMC stained with selected mAb (details in Table 1) as primary antibodies and stained with an isotype-specific R:PE conjugated secondary mAb, followed by directly conjugated mAbs CD16 and CD14. Following live/dead and singlets gating, PBMC were then gated based on the expression of CD16 and CD14 as CD14-CD16++, CD14+CD16+ and CD14+ CD16low/- as shown in Figure 2 and expression of the markers was then analysed. Geometric mean fluorescence intensity (MFI), corrected with the MFI of its corresponding FMO, for each of the molecules is shown in the table as an arithmetic mean and standard error -SE (n = 4). Different letters denote significant difference in marker expression levels (MFI) between the three myeloid populations (p < 0.05 (lower case) and p < 0.001 (upper case)).