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Table 1. Analytical characterization of synthetic peptides derived from Dsg3 and BP180Sequence M(av) calc M(av) measaR t /min bDsg3 peptides#196 PFGIFVVDK 104 1020.2 1020.2 23.0#2100 FVVDKNTGD 108 993.1 993.1 13.3#3104 KNTGDINIT 112 974.1 974.5 14.9BP180 peptides#4502 LERIRRSILPYGDS 515 1673.9 1674.2 18.6#5504 RIRRSILPYGDS 515 1431.6 1431.7 24.2a ESI-MS, Bruker Esquire 3000+b RP-HPLC, Knauer, Phenomenex Jupiter C18, 250x5mm, 5µm column, =214nm,gradient: 10-75 B% in 35 min. Eluent A: H 2 O+0.1 v/v% TFA, eluent B: AcN: H 2 O =80:20(v/v) +0.1 v/v% TFA.The culture medium induced IFN- production was considered as negative control. Aspositive control, PHA and SEB were used (data not shown). After 20 or 90 hrs ofincubation with peptides, PBMC of healthy donors showed no significant IFN- production(Figure 1). (Data of donor 2 are not shown.)Our results suggested that 0.025 mM peptide concentration was sufficient to induceIFN-γ release on PBMC. According to our results, 0.05 mM peptide concentration, in caseof some peptides and some patients, proved cytotoxic to the isolated PBMC (data notshown).The Dsg 3 derived peptides caused increased IFN- release on the PBMC of both PVpatients to a different extent (Figure 1). After 90 hrs of incubation similar data wereobtained for donor D3, but in case of donor D4 IFN- release was undetectable from thesupernatant (data not shown).Out of the two BP-180 derived peptides only the 14-mer peptide502 LERIRRSILPYGDS 515 induced IFN- production on one patients’ PBMC.Synthetic peptides at 0.025 mM and 20 hrs of incubation proved to be efficient todistinguish between IFN- production of PBMC obtained from healthy controls and PVpatients using ELISA. Further and fine T-cell epitope structure analysis with carefullyselected synthetic peptides could be considered in the development of synthetic antigens forthe early detection of PV with optimized T-cell response provoking capacity.AcknowledgmentsThese studies were supported by OTKA K61518, NKTH-OTKA 68358, GVOP-3.2.1-2004-04-0005/3.0 and GVOP-3.2.1-2004- 04-0352/3.0. Number of ethical permission: TUKEB 74-75/1998.References1. Wick, G., Beutner, E.H. Immunology 16, 149-156 (1969).2. Hertl, M. Int. Arch. Allergy Immunol. 122, 91-100 (2000).3. Hertl, M., Eming, R., Veldman, C. J. Clin. Invest. 116, 1159-1166 (2006).4. Amagai, M., Klaus-Kovtun, V., Stanley, J.R. Cell 67, 869-877 (1991).5. Kárpáti, S., Amagai, M., Prussick, R., Cehrs, K., Stanley, J.R. J. Cell Biol. 122, 409-15 (1993).6. Veldman, C.M., Gebhard, K.L., Uter , W., Wassmuth, R., Grötzinger, J., Schultz, E., Hertl, M. J.Immunol. 172, 3883-3892 (2004).7. Jurcevic, S., Hills, A., Pasvol, G., Davidson, R.N., Ivanyi, J., Wilkinson, R.J. Clin. Exp. Immunol.105, 416-421 (1996).8. Bősze, Sz., Caccamo, N., Majer, Zs., Mező, G., Dieli, F., Hudecz, F. Biopolymers (Peptide Science)76, 467-476 (2004).477