RESUMEN
HPV in remission CA,so the disease can be cured.
RESUMEN
Objective To screen and identify the predicted epitopes of synthesized HLA-A*0201restricted CTL derived from HPVll E7 antigen.Methods Five HPVll E7 CTL epitope peptides and terramers consisting of HLA-A*0201 were selected by way of computer and synthesized by Sanquin company,including HPVllE7 7-15(TLKDIVLDL),15-23(LQPPDPVGL),47-55(PLTQHYQIL),81-89(DLLLGTLNI)and 82-90(LLLGTLNIV).These peptides binding to human peripheral blood-derived DCs were tested for their ability to activate T cells isolated from peripheral blood lymphocytes of HLA-A*0201 healthy individuals.the number of specific tetramer+CD8+T cells by flow cytometry,the level of the section of IFN-γ by ELISA,and the ability of the CTL to kill the target cells were observed.Results The immature DCs could be fully activated by all the five HPV11 E7 peptides.Peptide-loaded mature DCs were able to stimulate the epitope-specific T cells responses in vitro.An increased frequency(P<0.05)of T ceils specific for the E7 7-15 epitope compared to other epitopes of HPV11E7.The epitope-specific CTL of E7 7-15 induced by the activated DCs specifically killed HPV11E7 expressing 293 cell line,and in a ratio of 50:1,the specific cytolytic activity was the strongest than the others(P<0.05).Conclusion DCs loaded with HPV11 E7 7-15(TLKDIVLDL)peptide can induce highly effective and specific ectogenic processed epitopespecific CTL responses in vitro.This peptide may be the candidate for development of CTL based vaccine in the treatment of HPV infeetions.
RESUMEN
Objective To investigate specific cellular immune response induced by dendritic cells(DCs) activated by human papilloma virus (HPV) 16 E7 peptide. Methods DCs separated from peripheral blood mononuclear cells were induced with granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4), the DCs were then activated by HPV 16 E7 peptide. After that the surface antigens on the DCs were detected by flow cytometer. Homogenic mixed lymphocyte reaction (HMLR) with DCs was performed. LDH release assay was also used to detect the activity of cytotoxic T lymphocytes(CTL)to Caski tumor cells, which was induced by DCs. Results The surface antigens on the activated DCs were highly expressed, such as CD1a(58.4 ? 6.7)%, CD80(70.6 ? 3.4)% and HLA-DR(74.8 ? 4.2)%. HMLR showed that the activated DCs stimulated T cell proliferation significantly. Antigen-specific CTL induced by the activated DCs specifically killed Caski tumer cells, but had no effect on SiHa and AN3CA cells. Conclusions DCs loaded with HPV 16 E7 peptide can induce highly effective and specific cellular immune response.
RESUMEN
Objective To introduce the amino acid substitution for HP V16E749-57(HLA-A2-restricted CTL epitope) and to identify the novel epitopes.Me thods Quantitative method was used to evaluate the affinity of the substituted peptides.To determine the peptide candidates to be synthesized and identified,the molecular models of the HLA-A2-peptide complex and CTL epitope candidates b ound to the HLA-A2 molecule were established by computer molecular modeling.Pep tides were synthesized and purified with standard Fmoc assay,lactate dehydrogen ase (LDH) release assay was used to determine their abilities of inducing the ge neration of specific CTLs.Results Modified peptides met the requirements of H LA-A2-restricted CTL epitopes.Peptide RLHYNIVTF had the abilitiy of inducing th e generation of specific CTLs.Conclusions Compared with HPV17E749-57 the mod ified peptide RLHYNIVTF has a higher antigenicity and affinity to HLA-A2.So,pe ptide RLHYNIVTF may be used as one of the HLA-A2-restricted candidate epitopes,instead of HPV17E749-57,for peptide vaccine in the treatment of HPV infection.
RESUMEN
Objective To screen and identify the predicted epitopes of synthesized predicted HLA-A2-restricted cytotoxic T lymphocytes (CTLs) derived from HPV16 E7 antigen. Methods The predicted epitopes of HLA-A2-restricted CTLs derived from HPV16 E7 antigen were synthesized and purified with Standard Fmoc assays, and the standard 51Cr release assay was used to determine their activities to induce specific CTL. Results Two epitopes of HLA-A2-restricted CTLs, namely E711-19 (YMLDLQPET) and E749-57 (RAHYNIVTF) derived from HPV16 E7 antigen were identified. Conclusion E711-19 (YMLDLQPET) and E749-57 (RAHYNIVTF) have antigenicity, and may be the candidates for development of peptide vaccine in the treatment of HPV infections.