Evaluation of auricular lymph node cell lymphocyte proliferation and cytokine production as non-radioactive endpoints during murine contact allergy

Figures & data

Table 1.  Change in ear thickness, lymph node weight, and cell count after treatment of mice with different concentrations of DNCB in AOO.

Labeling	Chemical	Vehicle	Concentration (% w/v)	Lymph node weight (g)	Cell count (10^7/ml)	Change of ear thickness (mm)	DI
In vivo	DNCB	AOO	—	0.0180 ± 0.0006	1.000 ± 0.082	0	> 1
			0.025%	0.0200 ± 0.0008	1.600 ± 0.082	0	> 1
			0.05%	0.0280 ± 0.0019*	2.400 ± 0.082*	0.010 ± 0.002*	> 1
			0.1%	0.0300 ± 0.0008*	2.800 ± 0.163*	0.030 ± 0.002*	> 1
			0.25%	0.0360 ± 0.0010*	3.000 ± 0.490*	0.030 ± 0.004*	> 1
Ex vivo	DNCB	AOO	—	0.010 ± 0.004	0.70 ± 0.24	0	> 1
			0.025%	0.012 ± 0.003	0.80 ± 0.16	0	> 1
			0.05%	0.020 ± 0.004*	1.40 ± 0.08*	0.020 ± 0.004*	> 1
			0.1%	0.030 ± 0.004*	1.60 ± 0.21*	0.030 ± 0.001*	> 1
			0.25%	0.038 ± 0.002*	1.80 ± 0.13*	0.040 ± 0.006*	> 1

Each value represents the mean ± SD.

*P < 0.01; Significant differences from vehicle control (Dunnett’s test).

Table 2.  OD₄₅₀, SI, and EC₃ values from the in vivo and ex vivo BrdU labeling studies.

Labeling	Chemical	Vehicle	Concentration (%w/v)	OD450	SI	EC3(%)
In vivo	DNCB	AOO	—	0.230 ± 0.012	1.00	0.059%
			0.025%	0.310 ± 0.018	1.34
			0.05%	0.690 ± 0.022	3.00
			0.1%	1.154 ± 0.025	5.00
			0.25%	1.580 ± 0.020	6.80
Ex vivo	DNCB	AOO	—	0.054 ± 0.010	1.00	0.054%
			0.025%	0.104 ± 0.016	2.00
			0.05%	0.152 ± 0.018	3.00
			0.1%	0.236 ± 0.012	4.60
			0.25%	0.329 ± 0.036	6.40

The OD₄₅₀ values shown are in terms of mean ± SD.

Table 3.  IL-2 levels in cultures of auricular lymph node cells from mice treated daily (3 days) with DNCB.

DNCB (in AOO) concentration (% w/v)	In vivo (+) PHA	SI in vivo (+) PHA	In vivo ( − ) PHA	Ex vivo (+) PHA	SI ex vivo (+) PHA	Ex vivo (−) PHA
0	80.65 ± 11.07^a	1.00	ND	7.89 ± 1.52	1.00	ND
0.025%	112.31 ± 13.36	1.39	ND	13.73 ± 1.53	1.74	ND
0.05%	228.99 ± 50.22*	2.84	ND	67.04 ± 15.10*	8.50	19.46 ± 3.75
0.1%	329.64 ± 65.89*	4.09	ND	97.54 ± 1.35*	12.36	79.13 ± 29.54
0.25%	593.15 ± 30.20*	7.35	ND	235.87 ± 66.40*	29.89	122.25 ± 16.62

^aIL-2 levels in terms of pg/ml (mean ± SE). *P < 0.01; Significant differences from vehicle control (Dunnett’s test).

ND: Not detectable.

Table 4.  IFNγ levels in cultures of auricular lymph node cells from mice treated daily (3 days) with DNCB.

DNCB (in AOO) concentration (% w/v)	In vivo (+) PHA	SI in vivo (+) PHA	In vivo ( − )PHA	Ex vivo (+) PHA	SI ex vivo (+) PHA	Ex vivo (−) PHA
0	266.06 ± 57.89^a	1.00	ND	88.28 ± 8.05	1.00	ND
0.025%	585.66 ± 191.48^†	2.20	ND	117.57 ± 15.21^†	1.33	45.5 ± 9.49
0.05%	912.54 ± 198.70*	3.43	6.23 ± 1.54	382.42 ± 25.13*	4.33	81.77 ± 8.34
0.1%	1358.8 ± 132.38*	5.11	27.12 ± 13.84	516.28 ± 32.33*	5.85	277.26 ± 25.60
0.25%	1539.07 ± 20.16*	5.78	272.18 ± 15.69	1378.37 ± 121.17*	15.61	682.47 ± 12.94

^aIFNγ levels in terms of pg/ml (mean ± SE).

^†P < 0.05. *P < 0.01; Significant differences from vehicle control (Dunnett’s test).

ND: Not detectable.

Table 5.  IL-4 levels in cultures of auricular lymph node cells from mice treated daily (3 days) with DNCB.

DNCB (in AOO) concentration (% w/v)	In vivo (+) PHA	SI In vivo (+) PHA	In vivo ( − ) PHA	Ex vivo (+) PHA	SI Ex vivo (+) PHA	Ex vivo (−) PHA
0	24.76 ± 4.65^a	1.00	ND	9.36 ± 2.29	1.00	ND
0.025%	29.00 ± 1.48	1.17	1.52 ± 0.61	13.98 ± 1.19	1.49	6.06 ± 3.35
0.05%	32.37 ± 5.14	1.31	4.65 ± 7.64	15.55 ± 0.69	1.66	8.33 ± 5.43
0.1%	62.62 ± 2.26*	2.53	7.64 ± 1.20	24.72 ± 2.79*	2.64	23.7 ± 1.22
0.25%	79.03 ± 8.24*	3.19	15.32 ± 1.16	44.16 ± 4.09*	4.72	27.92 ± 1.93

^aIL-4 levels in terms of pg/ml (mean ± SE).

*P < 0.01; Significant differences from vehicle control (Dunnett’s test).

ND: Not detectable.

Table 6.  IL-5 levels in cultures of auricular lymph node cells from mice treated daily (3 days) with DNCB.

DNCB (in AOO) concentration (% w/v)	In vivo (+) PHA	SI In vivo (+) PHA	In vivo ( − )PHA	Ex vivo (+) PHA	SI Ex vivo (+) PHA	Ex vivo (−) PHA
0	90.65 ± 43.41^a	1.00	ND	67.11 ± 2.34	1.00	ND
0.025%	280.19 ± 21.50	3.09	8.21 ± 4.52	150.26 ± 27.17	2.24	33.5 ± 14.14
0.05%	305.44 ± 37.36	3.37	25.38 ± 10.43	186.94 ± 27.17	2.78	57.76 ± 9.01
0.1%	510.43 ± 44.14*	5.63	55.6 ± 4.52	340.35 ± 27.46*	5.07	199.25 ± 9.36
0.25%	806.14 ± 61.28*	8.89	223.28 ± 25.65	560.19 ± 77.73*	8.35	354.23 ± 32.33

^aIL-5 levels in terms of pg/ml (mean ± SE).

*P < 0.01; Significant differences from vehicle control (Dunnett’s test).

ND: Not detectable.