Phenotypic and functional analysis of monocyte populations in cattle peripheral blood identifies a subset with high endocytic and allogeneic T-cell stimulatory capacity

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	<10^a	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⁺ CD16^low/- 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)).

ISSN: 1297-9716