. 2019 Apr 22;38(2):38–59. doi: 10.1089/mab.2018.0043

Table 10.

Experimental Analysis of Proliferation of CD3⁺/CD8⁺ T Cells with Anti-HLA-E Monospecific mAbs (TFL-033, TFL-034, TFL-073, TFL-074, TFL-145) and an Anti-HLA-E Polyreactive mAb (TFL-007) as the Control

Dilutions	No mAb (n = 5)
	CD3⁺ native cells					CD3⁺ lymphoblasts
		No PHA		With PHA		No PHA				Total	With PHA				Total
		CD4⁺/CD8⁻	CD4⁺/CD8⁻	CD4⁺/CD8⁻	CD4⁻/CD8⁺	CD4⁺/CD8⁻	CD4⁻/CD8⁺	CD4⁺/CD8⁺	CD4⁻/CD8⁻	Total	CD4⁺/CD8⁻	CD4⁻/CD8⁺	CD4⁺/CD8⁺	CD4⁻/CD8⁻	Total
	M	3063	547	1249	475	197	65	141	52	454	867	325	128	289	1609
	SD	149	86	99	37	33	14	35	15	70	115	126	43	84	267
	p²	NS	NS	<0.0001	NS	NS	NS	NS	NS	NS	0.001	0.001	NS	0.001	0.001
TFL-033 (IgG1) (n = 3) monospecific peptide (epitope) inhibition tested (tested dilution 1/3)
1/30	M	3185	755	1170	536	223	163	153	99	414	1129	505	152	412	2197
	SD	180	146	58	12	40	27	80	13	120	86	23	16	20	139
	p²	NS	NS	NS	0.009	NS	0.015	NS	0.005	NS	0.01	0.016	NS	0.014	0.004
1/150	M	3238	681	1149	508	252	120	205	68	645	1266	572	157	412	2407
	SD	14	64	21	22	30	17	13	9	28	80	31	14	16	116
	p²	NS	NS	NS	NS	0.05	0.001	0.02	NS	0.003	0.001	0.003	NS	0.001	0.001
TFL-034 (IgG1) (n = 3) monospecific peptide inhibition not tested
1/10	M	3048	631	1140	437	213	114	182	71	580	801	322	117	207	1446
	SD	83	90	158	54	16	24	10	22	44	97	29	2	36	162
	p²	NS	NS	NS	NS	NS	0.005	0.003	NS	0.015	NS	NS	NS	NS	NS
1/50	M	3354	687	1098	445	229	121	186	104	640	1424	602	160	362	2547
	SD	132	26	26	17	35	20	14	14	45	142	39	9	24	197
	p²	0.03	0.009	0.012	NS	NS	0.002	0.02	NS	NS	0.007	0.009	NS	NS	0.001
TFL-073 (IgG1) (n = 3) monospecific peptide inhibition not tested
1/10	M	2850	440	1089	374	153	63	137	58	411	682	307	107	209	1285
	SD	82	14	132	36	23	1	15	19	14	78	33	8	40	152
	p²	NS	NS	NS	0.03	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS
1/50	M	3119	637	1053	437	239	120	208	82	543	1009	472	140	425	2046
	SD	72	53	80	49	25	14	24	15	47	105	60	16	119	287
	p²	NS	NS	0.02	NS	NS	0.001	0.02	0.03	0.004	NS	NS	NS	NS	NS
TFL-074 (IgG1) (n = 3) monospecific peptide inhibition not tested
1/10	M	2933	362	1252	515	214	106	201	66	587	840	363	141	278	1623
	SD	95	304	75	51	13	18	18	9	25	119	3	20	56	179
	p²	NS	NS	NS	NS	NS	0.006	0.03	NS	0.017	NS	NS	NS	NS	NS
1/50	M	3401	624	1193	521	178	72	130	55	435	751	361	74	257	1444
	SD	28	13	8	17	27	13	15	3	40	54	64	19	52	273
	p²	0.007	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS
TFL-145 (IgG1) (n = 3) monospecific peptide inhibition not tested
1/20	M	3349	728	1206	500	173	83	157	47	459	771	349	97	257	1474
	SD	169	50	58	85	18	6	27	2	43	61	14	6	43	36
	p²	NS	0.006	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS	NS
1/200	M	3200	527	1160	410	238	76	168	66	548	1118	480	118	282	1998
	SD	229	37	14	30	32	6	9	9	12	65	71	23	74	271
	p²	NS	NS	NS	0.038	NS	NS	NS	NS	NS	NS	0.05	NS	NS	NS
TFL-007 (IgG1) (n = 3) HLA-I polyreactive nonspecific anti-HLA-E mAb as control
1/10	M	2876	451	1183	444	164	63	145	52	424	676	317	100	222	1315
	SD	135	72	19	26	33	2	3	17	47	79	25	4	29	125
	p²	NS	NS	NS	NS	NS	NS	NS	NS	0.027	NS	NS	NS	NS	NS
1/50	M	3088	667	1075	491	230	107	193	80	610	892	443	122	339	1795
	SD	65	16	55	49	23	7	17	4	26	26	18	8	21	38
	p²	NS	0.018	0.013	NS	NS	0.0005	0.019	0.006	0.002	NS	NS	NS	NS	NS

Peripheral blood lymphocytes were obtained from a normal healthy volunteer. Both PHA-untreated and PHA-treated CD4⁻/CD8⁺ cells but not CD4⁺/CD8⁻ or CD4⁺/CD8⁺ or CD4⁻/CD8⁻ cells are induced proliferation only by anti-HLA-E monospecific mAbs. The experimental protocols are described in detail elsewhere.⁽¹²⁵⁾ Paired test was performed between No mAb values versus TFL-mAb corresponding values. Significant values in the column of CD4⁻/CD8⁺ are shown in bold.

Ig, immunoglobulin; mAb, monoclonal antibody; NS, nonsignificant.