Heat shock protein 70/peptide complexes: potent mediators for the generation of antiviral T cells particularly with regard to low precursor frequencies

Tischer, Sabine; Basila, Megan; Maecker-Kolhoff, Britta; Immenschuh, Stephan; Oelke, Mathias; Blasczyk, Rainer; Eiz-Vesper, Britta

doi:10.1186/1479-5876-9-175

Table 2 Antiviral T-cell frequencies generated by the different stimuli.

From: Heat shock protein 70/peptide complexes: potent mediators for the generation of antiviral T cells particularly with regard to low precursor frequencies

A. Percentage of CD8⁺ T cells^#
CD8⁺ T cells (day 0: 20.22 ± 1.89)
	day 7	day 14	day 21
NC	25.48 ± 1.93	33.80 ± 20.15	41.92 ± 23.91
HSP70	25.20 ± 2.50	36.52 ± 12.65	47.70 ± 28.96
CMVpp65_495-503 peptide	27.07 ± 1.22	61.43 ± 11.61	63.06 ± 25.79
HSP70/CMV-PC	27.27 ± 0.95	79.83 ± 11.36**	83.17 ± 5.30*
^#cells were gated on viable lymphocytes
B. Percentage of CD25 ⁺ , CD69 ⁺ , and CD137 ⁺ T cells ^□
CD25⁺ T cells (day 0: 1.29 ± 0.21)
	day 7	day 14	day 21
NC	0.17 ± 0.29	0.23 ± 0.10	0.08 ± 0.04
HSP70	0.13 ± 0.23	0.28 ± 0.15	0.18 ± 0.08
CMVpp65_495-503 peptide	0.77 ± 0.29	36.18 ± 13.12	5.40 ± 5.29
HSP70/CMV-PC	1.74 ± 0.81	38.25 ± 8.16	18.10 ± 0.57*
CD69⁺ T cells (day 0: 13.62 ± 4.70)
NC	0.10 ± 0.10	0.23 ± 0.10	0.18 ± 0.18
HSP70	0.10 ± 0.10	0.30 ± 0.14	0.24 ± 0.27
CMVpp65_495-503 peptide	0.73 ± 0.22	2.68 ± 1.18	3.68 ± 4.18
HSP70/CMV-PC	1.33 ± 0.59	2.18 ± 0.35	5.82 ± 7.71
CD137⁺ T cells (day 0: 2.39 ± 0.01)
NC	0.10 ± 0.10	0.10 ± 0.08	0.04 ± 0.05
HSP70	0.07 ± 0.06	0.15 ± 0.06	0.14 ± 0.09
CMVpp65_495-503 peptide	0.70 ± 0.17	12.77 ± 4.24	6.60 ± 1.39
HSP70/CMV-PC	1.50 ± 0.52*	25.10 ± 4.67*	14.15 ± 5.87*
^□cells were gated on CD8⁺ T cells
C. Percentage of CD25 ⁺ , CD69 ⁺ , and CD137 ⁺ T cells ^○
CD25⁺ T cells (day 0: 5.98 ± 0.33)
	day 7	day 14	day 21
NC	51.03 ± 44.41	49.46 ± 43.74	51.03 ± 56.44
HSP70	49.57 ± 43.27	48.62 ± 45.50	50.50 ± 55.42
CMVpp65_495-503 peptide	87.37 ± 2.90	72.90 ± 25.77	73.03 ± 45.76
HSP70/CMV-PC	89.50 ± 5.40	78.55 ± 24.43	72.40 ± 47.37
CD69⁺ T cells (day 0: 16.49 ± 10.89)
NC	5.45 ± 4.74	10.73 ± 4.10	2.63 ± 4.56
HSP70	5.85 ± 3.89	12.08 ± 6.67	6.17 ± 8.45
CMVpp65_495-503 peptide	84.30 ± 0.52	2.93 ± 1.31	0.50 ± 0.44
HSP70/CMV-PC	84.93 ± 12.00	3.07 ± 1.24	0.63 ± 0.58
CD137⁺ T cells (day 0: 11.86 ± 1.01)
NC	2.60 ± 2.40	4.23 ± 3.10	3.30 ± 1.87
HSP70	4.00 ± 3.82	8.45 ± 4.63	4.83 ± 3.61
CMVpp65_495-503 peptide	88.05 ± 3.04	15.45 ± 7.19	10.73 ± 8.39
HSP70/CMV-PC	92.45 ± 5.87	17.05 ± 14.79	11.45 ± 9.73

^○cells were gated on A2/CMV-pentamer-positive CD8⁺ T cells
We assessed the capacity of T-cell subpopulations from five A2/CMV-pentamer-positive healthy donors to generate antiviral T cells in the presence of HSP70, CMVpp65_495-503 peptide, or HSP70/CMV-PC by enumerating the following specific T cells by flow cytometry. (A) CD8⁺ T cells gated on viable lymphocytes, (B) CD25⁺, CD69⁺, and CD137⁺ cells gated on CD8⁺ T cells and (C) CD25⁺, CD69⁺, and CD137⁺ cells gated on A2/CMV-pentamer-positive CD8⁺ T cells. PBMCs cultured in the presence of HSP70-peptide-binding buffer were used as negative controls (NC). The results of 5 independent experiments using cells from medium responder are expressed as mean ± SD. Asterisks shown indicate only statistically significant differences between levels in CMVpp65_495-503 peptide- and HSP70/CMV-PC-stimulated cells (* p < 0.05, ** p < 0.01 or *** p < 0.001).

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