Table 2. Flow cytometry data of NK cell... | American Society of Hematology

Table 2.

Flow cytometry data of NK cell activation mediated by mAb variants (frequency of CD3-CD56^dim cells)

mAb	Variant	IFN-γ⁺		IFN-γ⁺/CD107a⁺		CD107a⁺		Total activation∗		P value†
mAb	Variant	RhD^Pos	RhD^Neg	RhD^Pos	RhD^Neg	RhD^Pos	RhD^Neg	RhD^Pos	RhD^Neg	P value†
Brad3	WT	15.20	1.18	12.09	0.85	27.40	3.91	25.08	1.95
	GASDALIE	45.51	2.68	37.88	2.11	60.80	6.28	63.07	4.75	.0023
	AFUC	41.47	1.07	35.12	0.72	59.52	5.03	60.48	4.50	.0040
	AFUC GASDALIE	44.18	2.76	37.55	2.24	60.92	10.33	62.13	6.90	.0028
	GRLR	0.43	1.19	0.21	0.35	1.61	1.75	−3.57	−0.05	.0171
Fog1	WT	13.30	0.60	10.59	0.31	26.85	1.77	24.18	0.00
	GASDALIE	43.50	0.87	38.50	0.58	63.82	3.65	63.42	2.85	.0002
	AFUC	43.49	0.60	38.23	0.44	64.30	3.26	64.13	2.00	.0001
	AFUC GASDALIE	45.36	2.06	39.12	1.35	66.20	7.25	66.98	9.55	<.0001
	GRLR	0.46	1.10	0.21	0.34	1.58	1.70	−3.57	−0.10	.0029
VRC03	WT	0.96	1.17	0.39	1.90	2.35	2.89	−2.50	0.10	—
	GASDALIE	0.66	1.64	0.38	2.60	2.76	4.67	−2.38	−0.20	—
	AFUC	2.23	0.58	0.90	0.67	3.66	1.66	−0.43	0.30	—
	AFUC GASDALIE	3.83	0.37	1.92	0.86	4.54	1.83	1.05	0.30	—
	GRLR	0.44	1.49	0.14	1.36	1.36	4.72	−3.75	−0.10	—
No Ab		1.43	0.54	0.68	0.30	3.76	1.67	—	—	—

mAb	Variant	IFN-γ⁺		IFN-γ⁺/CD107a⁺		CD107a⁺		Total activation∗		P value†
mAb	Variant	RhD^Pos	RhD^Neg	RhD^Pos	RhD^Neg	RhD^Pos	RhD^Neg	RhD^Pos	RhD^Neg	P value†
Brad3	WT	15.20	1.18	12.09	0.85	27.40	3.91	25.08	1.95
	GASDALIE	45.51	2.68	37.88	2.11	60.80	6.28	63.07	4.75	.0023
	AFUC	41.47	1.07	35.12	0.72	59.52	5.03	60.48	4.50	.0040
	AFUC GASDALIE	44.18	2.76	37.55	2.24	60.92	10.33	62.13	6.90	.0028
	GRLR	0.43	1.19	0.21	0.35	1.61	1.75	−3.57	−0.05	.0171
Fog1	WT	13.30	0.60	10.59	0.31	26.85	1.77	24.18	0.00
	GASDALIE	43.50	0.87	38.50	0.58	63.82	3.65	63.42	2.85	.0002
	AFUC	43.49	0.60	38.23	0.44	64.30	3.26	64.13	2.00	.0001
	AFUC GASDALIE	45.36	2.06	39.12	1.35	66.20	7.25	66.98	9.55	<.0001
	GRLR	0.46	1.10	0.21	0.34	1.58	1.70	−3.57	−0.10	.0029
VRC03	WT	0.96	1.17	0.39	1.90	2.35	2.89	−2.50	0.10	—
	GASDALIE	0.66	1.64	0.38	2.60	2.76	4.67	−2.38	−0.20	—
	AFUC	2.23	0.58	0.90	0.67	3.66	1.66	−0.43	0.30	—
	AFUC GASDALIE	3.83	0.37	1.92	0.86	4.54	1.83	1.05	0.30	—
	GRLR	0.44	1.49	0.14	1.36	1.36	4.72	−3.75	−0.10	—
No Ab		1.43	0.54	0.68	0.30	3.76	1.67	—	—	—

Brad and Fog1 are anti-RhD mAbs while VRC03 is a non-RhD mAb (negative control).

∗

Total activation was calculated with subtraction of No-Ab control background.

†

P values show the significance of total NK cell activation of mAb variant versus IgG1 format in each group. Statistical analyses were conducted using one-way ANOVA followed by Dunnett’s multiple-comparison test (GraphPad Prism v9). Data are average of 3 replicates. In each replicate, a pool of NK cells from 3-4 independent donors was used.

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