Regulatory and activated effector T cells in chronic hepatitis C virus: Relation to autoimmunity

Table 1 Primers for TNFRSF4(OX40), granzyme 2, CD69, CD4, CD25, FoxP3 and interferonγ genes

TNFRSF4OX 40	Forward: ′5 GCA ATA GCT CGG ACG CAA TCT 3′
DQ032625.1	Reverse: ′5 GAG GGT CCC TGT GAG GTT CT 3′
Granzyme 2	Forward: ′5 TAC CAT TGA GTT GTG CGT GGG 3′
NM_004131.4	Reverse: ′5 GCC ATT GTT TCG TCC ATA GGA GA 3′
CD69	Forward: ′5 GGT CAC CCA TGG AAG TGG TC 3′
NM_001781.2	Reverse: ′5 GAC TTC GGA CCA CAG AGC AG 3′
CD4 NM_001195017.2	Forward: ′5 CTG CAA GTT CTC ACA CCG TC 3′
	Reverse: ′5 CTA GAG TTG CCT GCT CTG CC 3′
CD25 IL2R NM_000417.2	Forward: ′5 GCT CTA CAC AGA GGT CCT GC 3′
	Reverse: ′5 AGC ACA ACG GAT GTC TCC TG 3′
FoxP3	Forward: ′5 CCC ATC CCC AGG AGT CTT G 3′
NG_007392.1	Reverse: ′5 ACC ATG ACT AGG GGC ACT GTA 3′
Interferon γ	Forward: ′5 ATG GTT GTC CTG CCT GCA AT 3′
NG_015840.1	Reverse: ′5 CTT GCT TAG GTT GGC TGC CT 3′

Table 2 Relative gene expression of Tregs and Teffs specific genes in the four groups

	Group I (naïve)	Group II (Peg/Riba)	Group III (ANA+)	Group IV (control)
Granzyme 2 Tregs	0.704 ± 0.039	0.603 ± 0.046	0.400 ± 0.042	0.489 ± 0.053
CD69 Tregs	0.647 ± 0.037	0.54 ± 0.049	0.306 ± 0.036	0.383 ± 0.043
FoxP3 Tregs	0.531 ± 0.033	0.444 ± 0.046	0.330 ± 0.039	0.39 ± 0.030
TNFRSF4OX 40	0.596 ± 0.047	0.688 ± 0.057	0.707 ± 0.05	0.482 ± 0.056
CD4	0.584 ± 0.034	0.677 ± 0.048	0.712 ± 0.042	0.528 ± 0.05
CD25	0.595 ± 0.039	0.684 ± 0.053	0.73 ± 0.053	0.495 ± 0.041
Interferon γ	0.466 ± 0.035	0.483 ± 0.035	0.522 ± 0.044	0.357 ± 0.038

ANA: Antinuclear antibody.

Table 3 Multiple comparisons of gene expression of Tregs markers in the four studied groups

	Group I naïve	Group II Peg/Riba	Group III ANA+	Group IV control
FOX3
Group I naïve		aP ≤ 0.05	dP ≤ 0.001	bP ≤ 0.01
Group II Peg/Riba	aP ≤ 0.05		aP ≤ 0.05	bP ≤ 0.01
Group III ANA+	dP ≤ 0.001	aP ≤ 0.05		dP ≤ 0.001
Group IV control	bP ≤ 0.01	bP ≤ 0.01	dP ≤ 0.001
CD69
Group I naïve		bP ≤ 0.01	bP ≤ 0.01	dP ≤ 0.001
Group II Peg/Riba	bP ≤ 0.01		bP ≤ 0.01	dP ≤ 0.001
Group III ANA+	bP ≤ 0.01	bP ≤ 0.01		dP ≤ 0.001
Group IV control	dP ≤ 0.001	dP ≤ 0.001	dP ≤ 0.001
Granzyme
Group I naïve		dP ≤ 0.001	dP ≤ 0.001	dP ≤ 0.001
Group II Peg/Riba	dP ≤ 0.001		bP ≤ 0.01	dP ≤ 0.001
Group III ANA+	dP ≤ 0.001	bP ≤ 0.01		dP ≤ 0.001
Group IV control	dP ≤ 0.001	dP ≤ 0.001	dP ≤ 0.001

ANA: Antinuclear antibody.

Table 4 Multiple comparisons of gene expression of Teffs markers in the four studied groups

	Group I naïve	Group II Peg/Riba	Group III ANA+	Group IV control
TNFRSF4OX 40
Group I naïve		bP ≤ 0.01	aP ≤ 0.05	aP ≤ 0.05
Group II Peg/Riba	bP ≤ 0.01		NS (P > 0.05)	bP ≤ 0.01
Group III ANA+	aP ≤ 0.05	NS (P > 0.05)		dP ≤ 0.001
Group IV control	aP ≤ 0.05	bP ≤ 0.01	dP ≤ 0.001
CD4
Group I naïve		bP ≤ 0.01	aP ≤ 0.05	aP ≤ 0.05
Group II Peg/Riba	bP ≤ 0.01		NS (P > 0.05)	aP ≤ 0.05
Group III ANA+	aP ≤ 0.05	NS (P > 0.05)		bP ≤ 0.01
Group IV control	aP ≤ 0.05	aP ≤ 0.05	bP ≤ 0.01
CD25
Group I naïve		bP ≤ 0.01	aP ≤ 0.05	aP ≤ 0.05
Group II Peg/Riba	bP ≤ 0.01		NS (P > 0.05)	bP ≤ 0.01
Group III ANA+	aP ≤ 0.05	NS (P > 0.05)		dP ≤ 0.001
Group IV control	aP ≤ 0.05	bP ≤ 0.01	dP ≤ 0.001
Interferon-γ
Group I naïve		bP ≤ 0.001	bP ≤ 0.001	aP ≤ 0.05
Group II Peg/Riba	bP ≤ 0.001		NS (P > 0.05)	aP ≤ 0.05
Group III ANA+	bP ≤ 0.001	NS (P > 0.05)		bP ≤ 0.01
Group IV control	aP ≤ 0.05	aP ≤ 0.05	bP ≤ 0.01

ANA: Antinuclear antibody; NS: Non significant.