Table 3.

Spleen cell phenotypes in female B6C3F1/N mice following 13-week inhalation of C60

	Control	C60 Fullerene (m2/m3)					Trend Analysis
		0.011	0.033	0.084	0.112	0.167
Absolute Values (× 106)
50 nm C60
Mass-Based Exp. Conc. (mg/m3)	0	-	0.5	-	2	-	-
Spleen Cells (×106)	174.0 ± 8.2	-	161.3 ± 7.0	-	144.8 ± 4.5*	-	p ≤ 0.01
a Ig+	112.5 ± 4.3	-	92.5 ± 4.0**†	-	85.4 ± 3.4**††	-	p ≤ 0.01
b CD3+	28.2 ± 1.7	-	22.5 ± 1.1*	-	23.1 ± 1.2*	-	p ≤ 0.05
c CD4+CD8−	18.4 ± 1.2	-	15.5 ± 0.8	-	15.8 ± 0.9	-	NS
d CD4−CD8+	7.1 ± 0.4	-	6.4 ± 0.2	-	6.1 ± 0.5	-	NS
e NK1.1+CD3−	2.3 ± 0.2	-	2.2 ± 0.2	-	1.9 ± 0.1	-	p ≤ 0.05
f MAC-3+	8.0 ± 0.3	-	8.2 ± 0.9	-	6.8 ± 0.4	-	p ≤ 0.05
1 μm C60
Mass-Based Exp. Conc. (mg/m3)	0	2	-	15	-	30	-
Spleen Cells (×106)	147.7 ± 7.0	151.2 ± 10.4	-	147.5 ± 6.5	-	150.0 ± 7.5	NS
a Ig+	107.6 ± 4.8	100.6 ± 5.9	-	99.2 ± 5.0	-	101.9 ± 5.3	NS
b CD3+	20.2 ± 1.4	18.7 ± 2.2	-	20.5 ± 1.7	-	19.6 ± 2.1	NS
c CD4+CD8−	13.6 ± 0.9	12.3 ± 1.9	-	13.5 ± 1.3	-	11.8 ± 1.3	NS
d CD4−CD8+	6.2 ± 0.3	5.4 ± 0.8	-	4.9 ± 0.5††	-	3.5 ± 0.4††	p ≤ 0.01
e NK1.1+CD3−	1.9 ± 0.1	1.8 ± 0.1	-	2.2 ± 0.2	-	1.9 ± 0.2	NS
f MAC-3+	7.2 ± 0.4	6.8 ± 0.2†	-	5.9 ± 0.4††	-	5.6 ± 0.5*††	p ≤ 0.01
Percent Values
50 nm C60
Mass-Based Exp. Conc. (mg/m3)	0	-	0.5	-	2	-	-
a Ig+	65.1 ± 1.8	-	58.0 ± 3.2††	-	59.1 ± 2.0††	-	p ≤ 0.05
b CD3+	16.3 ± 1.0	-	14.1 ± 0.9	-	16.0 ± 0.8	-	NS
c CD4+CD8−	10.6 ± 0.6	-	9.7 ± 0.5	-	10.9 ± 0.6	-	NS
d CD4−CD8+	4.1 ± 0.3	-	4.0 ± 0.2	-	4.2 ± 0.3	-	NS
e NK1.1+CD3−	1.4 ± 0.1	-	1.4 ± 0.1	-	1.3 ± 0.1	-	NS
f MAC-3+	4.6 ± 0.2	-	5.0 ± 0.5	-	4.7 ± 0.2	-	NS
1 μm C60
Mass-Based Exp. Conc. (mg/m3)	0	2	-	15	-	30	-
a Ig+	72.9 ± 1.0	67.0 ± 2.3*††	-	67.4 ± 2.5	-	68.1 ± 1.0**††	p ≤ 0.05
b CD3+	13.6 ± 0.5	12.2 ± 0.7†	-	13.8 ± 0.6	-	12.9 ± 0.9	NS
c CD4+CD8−	9.1 ± 0.2	7.9 ± 0.8†	-	9.1 ± 0.6	-	7.8 ± 0.6†	NS
d CD4−CD8+	4.2 ± 0.1	3.5 ± 0.3	-	3.3 ± 0.3	-	2.4 ± 0.2** ††	p ≤ 0.01
e NK1.1+CD3−	1.3 ± 0.1	1.2 ± 0.1	-	1.5 ± 0.1	-	1.3 ± 0.1	NS
f MAC-3+	4.9 ± 0.4	4.6 ± 0.3	-	4.0 ± 0.2	-	3.8 ± 0.3	p ≤ 0.01

Mass-Based Exp. Conc. = Mass-Based Exposure Concentration

Spleen cell subsets defined based strictly on the presence or absence of the surface markers tested:

^aB-lymphocyte;

^bT-lymphocyte;

^cHelper T-lymphocyte;

^dCytotoxic T-lymphocyte;

^eNatural Killer cell;

^fMacrophage

Values represent the mean (± SE) from 8 animals per group;

^*p < 0.05 and

^**p < 0.01 from control (mass basis);

^†p < 0.05 and

^††p < 0.01 from combined controls (surface area basis; calculated via methods described in Sayers et al. 2016)

Combined control groups were used for surface area basis comparisons; see Supplementary Table S1 for combined control means