Echinacea and anti-inflammatory cytokine responses: Results of a gene and protein array analysis

Figures & data

Figure 1.  Relative gene transcription of CXCL chemokines (ELR-) 9-13. BEAS- 2B cells, in 75 cm ² flasks, uninfected or infected with rhinovirus type 14, were treated with Echinacea extract E1 or E2, or medium only, for 18 h. Total RNA was extracted, converted into cDNA and hybridized to DNA arrays containing 13,816 genes in duplicate. Data analysis was carried out as described in Materials and Methods and the normalized values from three separate experiments were converted into ratios: V/C = virus/control; VE1/V = virus + E1/virus; VE2/V = virus + E2/virus; CE1/C = control + E1/control; CE2/C = control + E2/control.

Table 1.  CXCL chemokines.

Chemokine	Principal target cells	V/C	VE1/V	VE2/V	CE1/C	CE2/C
CXCL1 (GROα)	neutrophils	0.64	0.55	2.30	1.07	10.63
		(1.01)	(0.30)	(0.28)	(0.48)	(0.43)
CXCL2 (GROβ)	neutrophils	0.85	0.43	3.38	1.22	19.7
CXCL3 (GROγ)	neutrophils	1.50	2.48	27.4	11.0	45.5
CXCL5 (ENA78)	neutrophils	0.5	14.6	0.64	0.48	0.44
CXCL8 (IL-8)	Neutrophils,basophils	0.83	5.71	23.76	16.8	35.72
		(4.10)	(0.06)	(0.40)	(0.88)	(5.6)
CXCL10 (IP-10)	Activated T-cells	0.99	1.05	1.04	1.00	1.04
		(2.62)	(0.50)	(0.91)	(2.25)	(2.90)

Data are expressed as ratios, C = control untreated cells, V = virus infected cells, E1 and E2 = cells treated with Echinacea E1 or E2. The upper numbers represent gene array data, and the numbers in parentheses represent corresponding ratios from the cytokine antibody results.

Figure 2.  CXCL chemokines 1, 2, 3, 5 and 8 (ELR+), relative gene transcription. BEAS- 2B cells, in 75 cm² flasks, uninfected or infected with rhinovirus type 14, were treated with Echinacea extract E1 or E2, or medium only, for 18 h. Total RNA was extracted, converted into cDNA and hybridized to DNA arrays containing 13,816 genes in duplicate. Data analysis was carried out as described in Materials and Methods and the normalized values from three separate experiments were converted into ratios: V/C = virus/control; VE1/V = virus + E1/virus; VE2/V = virus + E2/virus; CE1/C = control + E1/control; CE2/C = control + E2/control.

Figure 3.  CCL chemokines 2-5, 11, 17 and 18. BEAS- 2B cells, in 75 cm² flasks, uninfected or infected with rhinovirus type 14, were treated with Echinacea extract E1 or E2, or medium only, for 18 h. Total RNA was extracted, converted into cDNA and hybridized to DNA arrays containing 13,816 genes in duplicate. Data analysis was carried out as described in Materials and Methods and the normalized values from three separate experiments were converted into ratios: V/C = virus/control; VE1/V = virus + E1/virus; VE2/V = virus + E2/virus; CE1/C = control + E1/control; CE2/C = control + E2/control.

Table 2.  CCL chemokines, chromosome 17.

Chemokine	Principal target cells	V/C	VE1/V	VE2/V	CE1/C	CE2/C
CCL2 (MCP-1)	Monocytes, T-cells, basophils	1.44	0.52	1.03	0.63	4.32
		(1.20)	(0.41)	(0.14)	(0.42)	(0.43)
CCL8 (MCP-2)	Monocytes, T-cells, basophils, eosinophils	7.94	1.02	0.40	5.57	1.72
CCL3 (MIP-1α)	same	5.06	2.73	5.0	19.54	5.39
		(1.72)	(0.47)	(0.93)	(1.74)	(2.82)
CCL4 (MIP-1β)	same	3.17	1.48	0.51	11.4	22.2
		(1.76)	(0.34)	(1.20)	(1.88)	(4.0)
CCL5 (RANTES)	same	1.45	0.14	0.18	3.70	1.04
		(3.10)	(0.70)	(1.19)	(4.24)	(4.80)
CCL11 (eotaxin)	eosinophils	0.36	3.15	1.16	0.67	0.70
		(2.27)	(0.63)	(1.05)	(2.20)	(4.70)
CCL17 (TARC)	Various T-cells	12.46	0.28	0.06	9.32	9.84
		_	_	_	_	_
CCL18 (PARC)	Various T-cells	66.3	0.01	0.52	0.02	2.62
		(1.40)	(0.67)	(1.60)	(1.80)	(2.70)
CCL23 (MPIF)	Monocytes T-cells	7.26	0.43	0.59	7.35	0.59
		_	_	_	_	_

C = control untreated cells, V = virus infected cells, E1 and E2 = cells treated with Echinacea E1 or E2. The upper numbers represent gene array data, and the numbers in parentheses represent corresponding ratios from the cytokine antibody results.

Table 3.  Interleukin-1 family.

IL member	Activities	V/C	VE1/V	VE2/V	CE1/C	CE2/C
IL1F1 (IL-1α)	Activation of T-cells & macrophages; fever	0.40	0.52	0.67	0.46	0.11
		(0.69)	(0.55)	(0.89)	(0.86)	(2.43)
IL1F2 (IL-1β)	same	0.40	1.90	1.11	0.57	0.25
		(1.86)	(0.40)	(1.20)	(1.40)	(2.93)
IL-1β convertase (caspase-1)		0.77	0.31	1.30	0.67	0.55
		_
IL1F3 (IL-1RA)	IL-1 receptor antagonist	3.90	8.0	0.15	12.6	1.80
		(2.2)	(0.31)	(0.88)	(1.30)	(2.80)
IL-1R1	IL-1 receptor	0.50	5.14	2.71	4.38	0.15
		_
IL1F4 (IL-18)	Induces IFNy; promotes Th-1 response	0.97	0.51	1.10	0.76	1.10
		_
IL1F7 (IL-1 ζ)	Inhibits IL-18 activity	0.50	–	0.35	0.22	0.59
		_
IL1F8 (IL-1 η)	Agonist of IL-1 receptor 2; upregulates IL-6/8	1.49	0.76	0.47	0.78	0.47
		_

C = control untreated cells, V = virus infected cells, E1 and E2 = cells treated with Echinacea E1 or E2. The upper numbers represent gene array data, and the numbers in parentheses represent corresponding ratios from the cytokine antibody results.

Table 4.  Cytokine cluster: chromosome 5q31.

Cytokine	Activities	V/C	VE1/V	VE2/V	CE1/C	CE2/C
IL-4 (interleukin-4)	Activation of B-cells; differentiation of Th-2 cells	4.53	0.89	0.25	1.35	0.72
		(1.20)	(1.06)	(1.68)	(1.95)	(1.70)
IL-5 (interleukin-5)	Eosinophil growth & differentiation	3.18	1.21	0.01	4.43	2.87
		(1.42)	(0.92)	(1.20)	(2.45)	(2.60)
IL-13 (interleukin-13)	B-cell growth & differentiation; involved in asthma/allergy	0.35	1.83	2.84	1.50	0.85
		_
GM-CSF (Granulocyte/ macrophage colony- stimulating factor)	Growth & differentiation of myelomonocyte lineage, particularly dendritic cells	0.19	1.35	5.0	0.44	1.20
		(2.24)	(1.66)	(1.86)	(2.65)	(6.05)

C = control untreated cells, V = virus infected cells, E1 and E2 = cells treated with Echinacea E1 or E2. The upper numbers represent gene array data, and the numbers in parentheses represent corresponding ratios from the cytokine antibody results.

Table 5.  Cytokines involved in inflammatory conditions.

Cytokine	Activities	V/C	VE1/V	VE2/V	CE1/C	CE2/C
IL-6 Interleukin 6	Growth/differentiation of T & B cells; fever; acute phase responses	0.86	1.08	1.04	0.60	0.46
		(5.50)	(0.29)	(0.44)	(2.76)	(9.80)
IL-10 Interleukin 10	Suppresses macrophage functions	1.05	0.56	0.97	0.59	0.93
		(2.30)	(0.96)	(2.06)	(4.50)	(5.10)
IL-12 Interleukin 12	Activates NK cells	1.45	0.69	0.68	1.71	1.20
		(1.10)	(1.20)	(1.50)	(1.78)	(1.50)
IL-16 Interleukin 16	Attracts CD4 T-cells, monocytes & eosinophils	8.11	0.18	0.03	7.30	0.13
		_
IL-17 Interleukin 17	Induces cytokine production in various cells	0.36	1.60	0.28	0.32	0.14
		(3.14)	(0.61)	(0.95)	(3.13)	(3.80)
TNFα Tumor necrosis factor	Inflammation	44.4	0.62	0.08	18.86	14.44
		(1.33)	(0.40)	(1.16)	(1.36)	(1.70)
FASL Fas ligand	Member of TNF family; apoptosis	2.30	1.80	0.15	2.70	1.50
		(2.30)	(0.71)	(0.70)	(2.70)	(3.06)
TGFβ Transforming growth factor β	Anti-inflammatory; inhibits cell growth; attracts mast cells	(1.82)	(0.63)	(1.08)	(1.74)	(2.65)

C = control untreated cells, V = virus infected cells, E1 and E2 = cells treated with Echinacea E1 or E2. The upper numbers represent gene array data, and the numbers in parentheses represent corresponding ratios from the cytokine antibody results.

Table 6.  Summary of cytokines showing modulation.

Cytokine induced by virus;Gene level	Modulated by E1/E2 or both	Cytokine induced by virus;protein level	Modulated by E1/E2 or both
CXCL11	both	CXCL8	both
CXCL12	E2	CXCL10	both
CCL8	E2	CCL3	E1
CCL3	no	CCL4	E1
CCL4	E1	CCL5	E1
CCL17	both	CCL11	E1
CCL18	both	IL1β	E1
CCL23	both	IL1RA	both
IL1RA	E1	GM-CSF	no
IL4	E2	IL6	both
IL5	E2	IL10	no
IL16	both	IL17	E1
TNF	both	FASL	both
FASL	E2