. 2018 May 1;163(2):639–654. doi: 10.1093/toxsci/kfy066

Table 1.

The Number of Immune Cells in Immunologically Naïve Female and Male Offspring

	Female 0.1 µg/ml versus Control			Male 0.1 µg/ml versus Control			F versus M	Female 1 µg/ml versus Control		Male 1 µg/ml versus Control		F versus M
	Control	0.1 µg/ml	p-Value^a	Control	0.1 µg/ml	p-Value^a	p-Value^b	1 µg/ml	p-Value^a	1 µg/ml	p-Value^a	p-Value^b
Bone marrow
Cell count	12433333 ± 742930	11148333 ± 1153077	.53	11850000 ± 640638	14412500 ± 781658	.07	.15	8305000 ± 150000	.05^*	14050000 ± 819807	.12	.01^*
HSC	81.7 ± 30.3	123 ± 26	.56	181 ± 47	233 ± 41	.63	.10	277 ± 40	.02^*	232 ± 44	.64	.56
LSK	1681.2 ± 114.3	1681 ± 651	.99	1617 ± 154	1746 ± 190	.24	.84	1249 ± 189	.76	1993 ± 148	.81	.04^*
LK	37727 ± 3013	38353 ± 9465	.99	40321 ± 1143	50790 ± 5188	.09	.27	33628 ± 192	.89	50129 ± 2483	.12	.01^*
MPP	1441.3 ± 111.7	1335 ± 565	.97	1253 ± 91	1207 ± 214	.97	.82	745 ± 185	.45	1560 ± 146	.33	.03^*
Lineage ^-	334137 ± 40143	284293 ± 89951	.81	313340 ± 23271	364988 ± 42847	.48	.41	305411 ± 16724	.94	404781 ± 32400	.15	.12
CLP	4561 ± 1431	4516 ± 2171	.99	4639 ± 446	4691 ± 778	.99	.93	156 ± 38	.25	4616 ± 163	.99	<.0001^*
GMP	3229.2 ± 555.1	4890 ± 444	.10	4417 ± 499	5646 ± 685	.23	.44	3727 ± 514	.76	5258 ± 349	.46	.07
Pre-GM	8716.2 ± 691.5	8785 ± 2383	.99	11272 ± 316	14981 ± 1787	.09	.09	5484 ± 265	.39	14625 ± 931	.13	.003^*
Pre-MegE	15717 ± 1315	14403 ± 3406	.89	14148 ± 331	17659 ± 1437	.10	.37	14119 ± 255	.88	16469 ± 1344	.31	.31
Thymus
Cell count	134700000 ± 15085534	7992833± 38156620	.31	86075000 ± 6312702	84687500 ± 4441583	.97	.89	198750000 ± 33250000	.29	77562500 ± 1852406	.36	.004^*
DN1	1029398 ± 128817	629804 ± 229799	.22	652118 ± 46528	633010 ± 37150	.95	.99	993675 ± 50325	.99	593431 ± 62092	.63	.02^*
DN2	154645 ± 17105	123131 ± 56585	.78	82867 ± 4305	80256 ± 4563	.95	.41	150895 ± 41665	.99	81100 ± 9931	.98	.08
DN3	1684308 ± 190926	1055676 ± 502342	.36	979296 ± 62343	968785 ± 21799	.97	.84	1995775 ± 324225	.79	933129 ± 14914	.63	.01^*
DN4	929283 ± 44022	1039598 ± 175690	.81	927516 ± 118930	867171 ± 69782	.88	.35	1045175 ± 300425	.83	812128 ± 111851	.65	.40
DP	64765250 ± 4519003	4214308± 21474537	.47	43490400 ± 3354243	43503338 ± 5616368	1	.95	79919750 ± 22160250	.75	41521763 ± 2323515	.92	.05
DP CD3⁺	37535 ± 4244	35116 ± 18488	.98	94271 ± 24827	66288 ± 9805	.45	.17	35117.5 ± 13603	.99	90209 ± 14613	.98	.08
TCRβ⁺	6829968 ± 476592	6791068 ± 2675537	.99	6917440 ± 462168	5960451 ± 41223	.34	.73	7544450 ± 1619550	.95	5804218 ± 724325	.25	.30
CD4⁺	5563755 ± 415844	4480870 ± 1974304	.79	4891533 ± 349205	4651036 ± 137026	.82	.92	7347550 ± 1886050	.62	4259121 ± 401819	.31	.08
CD8⁺	680505 ± 55178	930392 ± 96601	.16	885978 ± 206229	751826 ± 106954	.75	.29	533375 ± 185825	.53	723828 ± 105159	.67	.38
TCRγδ⁺	483158 ± 45652	313629 ± 138319	.33	378986 ± 27842	377249 ± 23029	.99	.62	526650 ± 30150	.93	395438 ± 69299	.95	.28
Treg	126705 ± 11419	120334 ± 25422	.97	109801 ± 9453	103152 ± 7950	.80	.49	151058 ± 36863	.69	88345 ± 7749	.18	.07
Spleen
Cell count	86737500 ± 1371298	8221666± 31853497	.98	8998750± 10669707	107612500 ± 8841907	.32	.06	76000000 ± 19500000	.93	117512500 ± 6614390	.10	.42
CD19⁺	42901875 ± 8244859	4150248± 17869820	.99	4446851± 10596693	57843813 ± 5629245	.38	.36	42249750 ± 1 3999750	.99	60730938 ± 9521355	.26	.17
CD4⁺	9179363 ± 1756623	10686967 ± 4435512	.91	11378904 ± 2552025	14578738 ± 865956	.33	.36	7227550 ± 1577550	.88	15235463 ± 969390	.22	.01^*
CD8⁺	6755044 ± 1108823	7373400 ± 2819035	.96	7579081 ± 1212533	9894250 ± 747637	.18	.36	4506000 ± 1042550	.68	9921128 ± 642899	.17	.01^*
NK1.1⁺	1864925 ± 286698	1721660 ± 705751	.97	2457671 ± 244963	2309921 ± 295240	.86	.43	1148275 ± 255575	.55	2769471 ± 80402	.54	.001^*
TCRγδ⁺	1404270 ± 224021	1332622 ± 564340	.99	1378785 ± 208629	1592971 ± 114013	.49	.62	1039450 ± 288000	.77	1663444 ± 65032	.31	.04^*
Treg	492888 ± 134031	572553 ± 240016	.93	758630 ± 230462	933366 ± 142882	.67	.23	441275 ± 45775	.97	995236 ± 64601	.5	.01^*
F4/80⁺	2736355 ± 362139	2959867 ± 701954	.94	2526675 ± 389486	3384780 ± 384762	.22	.59	3851600 ± 761050	.38	3708730 ± 308728	.08	.84
Gr-1⁺	4945213 ± 691402	5847227 ± 1601634	.79	5473478 ± 1065955	7106785 ± 546481	.26	.44	5581725 ± 960025	.91	7565911 ± 490051	.14	.10
Gr-1⁺CD11b⁺	449216 ± 65317	889747 ± 275714	.21	374870 ± 33306	469855 ± 33816	.14	.13	1457325 ± 242575	.02^*	542254 ± 34726	.01^*	.004^*
CD11c⁺MHCIIhi	1196778 ± 203674	1127800 ± 425883	.98	1103168 ± 222118	1405939 ± 137023	.37	.51	915300 ± 163850	.78	1501859 ± 119692	.21	.04^*
CD11b⁺DCs	135978 ± 38691	146450 ± 46234	.98	140680 ± 22079	180088 ± 4949	.17	.43	80218 ± 12418	.61	196008 ± 13632	.54	.01^*
CD103⁺DCs	21814 ± 5846	29950 ± 12584	.77	27152 ± 7706	27782 ± 9831	.99	.90	20668 ± 12758	.99	40007 ± 9565	.53	.30
Lymph nodes
Cell count	906250 ± 356587	1246333 ± 230860	.88	1205250 ± 142966	1281250 ± 248510	.95	.92	1750500 ± 1279500	.57	665000 ± 86920	.12	.24
NK1.1⁺	339467 ± 152821	376573 ± 163194	.99	324370 ± 20897	418592 ± 103462	.48	.81	726215 ± 510026	.49	194269 ± 20803	.28	.18
CD19⁺	184994 ± 89183	206919 ± 78508	.99	195106 ± 16840	235362 ± 90192	.82	.83	428606 ± 319805	.45	102250 ± 17897	.41	.17
TCRγδ⁺	131612 ± 57008	166186 ± 59364	.95	143302 ± 13848	195615 ± 78622	.66	.37	293009 ± 219062	.46	74033 ± 13529	.49	.24
CD4⁺	3744 ± 1433	4361 ± 1424	.98	4831 ± 845	4884 ± 1376	.99	.83	8238 ± 6306	.45	1909 ± 206	.09	.17
CD8⁺	18441 ± 10365	22114 ± 7900	.95	29839 ± 11946	14580 ± 3132	.27	.79	18404 ± 13411	1	6903 ± 997	.08	.18
Treg	13911 ± 6572	13545 ± 4531	.99	16110 ± 3418	22151 ± 10075	.73	.52	34468 ± 26132	.39	8232 ± 1871	.59	.18
F4/80⁺	6276 ± 2376	11119 ± 2556	.55	6332 ± 439	7071 ± 773	.61	.14	10887 ± 8202	.65	3314 ± 540	.01^*	.21
Gr-1⁺	51600 ± 20201	74254 ± 23100	.91	62885 ± 6323	82352 ± 29324	.66	.85	163213 ± 117668	.25	31898 ± 5218	.39	.14
Gr-1⁺CD11b⁺	624 ± 266	1992 ± 1385	.45	524 ± 85	613 ± 196	.84	.30	1766 ± 871	.63	251 ± 33	.26	.04^*
CD11c⁺MHCII^hi	7617 ± 3016	11403 ± 3023	.79	7074 ± 193	9223 ± 2031	.38	.56	15758 ± 10906	.47	4117 ± 376	.19	.16
CD11b⁺DCs	1933 ± 679	4089 ± 1484	.35	1610 ± 292	2094 ± 144	.18	.17	2521± 1721	.93	1120 ± 68	.17	.26
CD103⁺DCs	1729 ± 749	2358 ± 1345	.91	1540 ± 177	1925 ± 613	.69	.76	3302± 2455	.66	718 ± 85	.25	.16

The table depicts the mean number (±SEM) of the indicated cell population in primary and secondary immune organs from immunologically naïve male and female offspring at maturity (6 weeks of age). There were 4–8 mice of each sex per group, and same-sex offspring within each group were from different dams. Bone marrow, thymus, and peripheral lymph nodes (PLN) were isolated and cell populations identified using multiparametric analytical flow cytometry. Cells enumerated in the bone marrow include: HSC (Lineage[Lin])^negSca-1⁺cKit⁺CD105^hiCD150⁺); LSK cells (Lin^negSca-1⁺Kit⁺ cells); LK cells (Lin^negKit⁺ cells); multipotent progenitors (MPP; Lineage^negSca-1⁺cKit⁺CD105^intCD150^neg); common lymphocyte progenitors (CLP; Lineage^negSca-1⁺cKit⁺CD105^hiCD150⁺CD127⁺); granulocyte monocyte progenitors (GMP; Lin^negKit⁺CD16/32⁺CD150^neg); pre-granulocyte/monocyte cells (pre-GM; Lin^negKit⁺CD16/32^negCD150^negCD105^int); pre-megakaryocyte/erythrocyte cells (pre-MegE; Lin^negKit⁺CD16/32^negCD150⁺CD105^hi). In the thymus, cells were defined as double negative: DN1 (CD44⁺CD25^negCD4^negCD8^neg); DN2 (CD44⁺CD25⁺CD4^negCD8^neg); DN3 (CD44^negCD25⁺CD4^negCD8^neg); DN4 (CD44^negCD25^negCD4^negCD8^neg); double positive (DP; CD3⁺CD4⁺CD8⁺); CD8 SP (CD3⁺CD4^negCD8⁺); CD4 SP (CD3⁺CD4⁺CD8^neg); Treg (Foxp3⁺CD25⁺CD4⁺CD8^negCD3⁺); γδT cells (γδTCR⁺CD3⁺CD4^-CD8^-). In the lymph node, cells were defined as: CD8⁺ T cells (CD3⁺CD8⁺); CD4⁺ T cells (CD3⁺CD4⁺); Treg (Foxp3⁺CD25⁺CD4⁺); neutrophils (Gr-1⁺); macrophages (F4/80⁺); dendritic cells (DCs; CD11c⁺MHCII^hi), CD11b⁺DCs (CD11b⁺CD103⁻CD11c⁺MHCII^hi cells) and CD103⁺DCs (CD11b⁻CD103⁺CD11c⁺MHCII^hi cells); γδT cells (γδTCR⁺CD3⁺CD4⁻CD8⁻); NK cells (CD3^negNK1.1⁺); B cells (CD3^negCD19⁺).

The p-value for mixture-exposed group compared with control group within the indicated sex.

The p-value for female versus male offspring from dams given indicated concentration of the mixture.

Demarcates p ≤.05.