Screening NK-, B- and T-cell phenotype and function in patients suffering from Chronic Fatigue Syndrome

Table 1 Antibody combinations used for the phenotypic characterization of T, NK, B cells and for the determination of absolute counts

Antibody combinations/							Fluorochrome
CELL TYPE	Phenotype	Tube #	V450/Pacific blue	V500/Am Cyan	FITC	PE	APC/Alexa Fluor 647	PerCP/PerCP-Cy5	PE-Cy7	APC-cy7
T cells	Thymic output/Exhaustion	T1			CD95	PD-1	CD4	HLA-DR	CD8	CD3
		T2			CD45RA	CD31	CD4	CD38^d	CD8	CD3
		T3					CD4		CD8	CD3
	CD8 activation	T4	CD4		HLA-DR		CD45RO	CD38^d	CD8	CD3
	CD8 activation	T5	CD4						CD8	CD3
	Main subsets/senescence	T6	CD45RA	CD8	CD57^e	CD28	CD27	CD4	CCR7	CD3
	Main subsets/senescence	T7		CD8				CD4		CD3
	Treg/Prolif./Anergy	T8	CD4	CD8	KI67	FOXP3^e	CD127^c	CD5^d	CD25	CD3
	Treg/Prolif./Anergy	T9	CD4	CD8	IRR IgG	IRR IGG2a^e				CD3
NK cells	Activation/receptor profile	NK1	CD69	CD3^b/CD19^b	CD107a	Nkp44	CD57^e	CD25	CD56	CD16
		NK2	NKp46^a,ed	CD3^b/CD19^b	CD107a	NKG2D^e	Nkp30 ^c	NKG2A^e	CD56	CD16
		NK3		CD3^b/CD19^b					CD56	CD16
B cells	Mature/Memory/Transitional	B1			CD1c	IgD^e	IgM	CD38	CD27^e	CD19
		B2			CD23	IgD^e	CD5	CD38	CD10^e	CD19
		B3				IgD^e				CD19
	Switched memory	B4			CD1c^e	IgG^e	IgA	CD38	CD27^e	CD19
	Switched memory	B5				IgG^e	IgA^e			CD19

^a Pacific blue-coupled antibody.
^b AmCyan-coupled antibody.
^c Alexa Fluor 647-coupled antibody.
^d PerCP-Cy5.5 coupled antibody.
^e sources different from BD Biosciences (NKp46 and CD57 from BioLegend; NKG2A from R&D Systems, NKG2D and CD1c from Santa Cruz; FOXP3, IRR IGG2a, CD27 and CD10 from eBiosciences; IgG and IgA from Jackson ImmunoResearch, CD57 from Beckman Coulter).

ISSN: 1479-5876