Analysis of peptide‐binding motifs for two disease associated HLA‐DR13 alleles using an M13 phage display library

Abstract

SUMMARYMajor histocompatibility complex (MHC) molecules bind peptides bearing an appropriate 'sequence motif' for MHC binding. The use of phage display libraries exploits the ability of MHC class II molecules to exchange peptides in solution and thus select out peptide sequences with high-affinity binding from a large array of random peptides. We have analysed the peptide binding motifs of HLA-DRB1*1301 and *1302 using affinity purified HLA-DR13 molecules to purify sequentially HLA-DR13-binding peptides from a large random library of M13 phage containing nonamer inserts in the pIII coat protein. These DR13 alleles differ only at position 86 of the HLA-DRb chain, where they contain valine and glycine residues respectively. These alleles were chosen because of their association with protection from severe malaria and chronic hepatitis B virus infection in West Africa. Analysis of the phage bound to these DR molecules suggests binding motifs. We compare the results derived from the use of the phage display library with results obtained from analysis of eluted peptides and peptidebinding studies. This analysis shows that although there is a common theme to motifs derived using different methods, there are also subtle variations between them.

Functional classification of class II human leukocyte antigen (HLA) molecules reveals seven different supertypes and a surprising degree of repertoire sharing across supertypes

“…A similar preference for acidic or polar residues in positions 4 and or 6 has also been suggested for DRB3 molecules (Verreck et al 1996). A preference for basic residues has been described for DRB1*1302 (Boitel et al 1995; Davenport et al 1995, 1996; Verreck et al 1996) and DRB3*0101 (Verreck et al 1996). A detailed motif for DRB4*0101 has not been described, but a preference for basic residues in some positions has been suggested (Kobayashi et al 1996).…”

Immunisation with recombinant modified vaccinia virus Ankara expressing HIV‐1 gag in HIV‐1‐infected subjects stimulates broad functional CD4+ T cell responses

“…3B), as is illustrated by the effects of truncation of the N-terminal Phe and C-terminal Ala on the responses to FKTLRAEQATQEVKNW and RDYVDRFFKTLRAEQA, respectively. This suggested that the 9-mer FKTLRAEQA, which fits the predicted peptide-binding motif for HLA-DRB1*1301 and *1302 [29], represented the core binding region of two distinct epitopes in this region. The p24 163-177 epitope defined by Malhotra et al was also recognised by CD4 + T cells from this subject [28].…”

Understanding the Host Genetics of Chronic Hepatitis B and C

“…Elution and binding studies suggest that HLA-DRB1*1302 is a relatively promiscuous peptide binder in comparison to other HLA class II molecules. 20,21 The ability to bind and present a wide range of epitopes is likely to generate a polyclonal and multispecific T-helper cell response, as seen in individuals who spontaneously eliminate the infection. Furthermore, a broad range of potential T cell epitopes would reduce the opportunity for the virus to evade recognition through sequence variation.…”