Collection of phage–peptide probes for HIV-1 immunodominant loop-epitope
- 作者：Yadira Palacios-Rodrí ; guez ; Tatiana Gazarian ; Merrill Rowley ; Abraham Majluf-Cruz ; Karlen Gazarian
- 关键词：Biopanning ; Epitopes ; gp41 ; HIV-1 ; Mimotopes ; Phage-display
- 刊名：Journal of Microbiological Methods
- 出版年：2007
- 期刊代码：59_01677012
- 类别：imm
- 出版时间：February 2007
- 卷：68
- 期：2
- 页码：225-235
- 文件大小：453 K
摘要
Early diagnosis and prevention of human immunodeficiency virus type-1 (HIV-1) infection, which remains a serious public health threat, is inhibited by the lack of reagents that elicit antiviral responses in the immune system. To create mimotopes (peptide models of epitopes) of the most immunodominant epitope, CSGKLIC, that occurs as a loop on the envelope gp41 glycoprotein and is a key participant in infection, we used phage-display technology involving biopanning of large random libraries with IgG of HIV-1-infected patients. Under the conditions used, library screening with IgG from patient serum was directed to the CSGKLIC epitope. Three rounds of selection converted a 12 mer library of 109 sequences into a population in which up to 79%of phage bore a family of CxxKxxC sequences (“x” designates a non-epitope amino acid). Twenty-one phage clones displaying the most frequently selected peptides were obtained and were shown to display the principal structural (sequence and conformational), antigenic and immunogenic features of the HIV-1 immunodominant loop-epitope. Notably, when the mixture of the phage mimotopes was injected into mice, it induced 2- to 3-fold higher titers of antibody to the HIV-1 epitope than could be induced from individual mimotopes. The described approach could be applicable for accurately reproducing HIV-1 epitope structural and immunological patterns by generation of specialized viral epitope libraries for use in diagnosis and therapy.