Fine mapping of HLA class II monoclonal antibody specificities using transfected L cells

Altmann, Daniel M.; Heyes, J. M.; Ikeda, Hitoshi; Sadler, A. M.; Wilkinson, David; Madrigal, J. Alejandro; Bodmer, Julia G.; Trowsdale, John

doi:10.1007/bf01787329

Cited by 27 publications

(17 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…13 The DRB1*1301 and *1302 gene products were then purified from the DRB3 products using the TAL15 . 1 monoclonal antibody 14 (obtained from S. Marsh and J. Bodmer). The purified DR13 molecules were then biotinylated.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 1996

View full text Add to dashboard Cite

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.

show abstract

Section: Methodsmentioning

confidence: 99%

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

et al. 1996

View full text Add to dashboard Cite

show abstract

“…A low-density-enriched fraction was subsequently prepared from thawed thymus cell suspension (12) For some experiments we used as APCs the Epstein-Barr virus-induced (EBV) cell lines listed in Table 1 and, for others, mouse L cells transfected with the Mulcos expression vector carrying multiple copies of DRA and DRB1* 0401 full-length cDNA clones and selected for stable high class II expression (24,25). To prevent autopresentation of peptides by these class II-positive T cells to each other (26), 5 x 106…”

mentioning

confidence: 99%

Critical role for the Val/Gly86 HLA-DR beta dimorphism in autoantigen presentation to human T cells.

Ong

Willcox

Wordsworth

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Helper T lymphocytes recognize fragments of foreign (or self) antigens in the peptide-binding clefts of major histocompatibility complex class II molecules; their activation is a crucial step in the induction of many immune and autoimmune responses. While studying the latter, we raised a T-cell line from the thymus of a myasthenia gravis patient against recombinant a subunit of the human acetylcholine receptor, the target of this autoimmune disease. The line responds to the 144-156 region of the human sequence and not to the same region of the electric fish homolog, which differs by only three residues. These CD4' T cells recognize this epitope only in the context of HLA-DR4 class II molecules, of which the variants with Gly86 are absolutely required. Thus the naturally occurring alternatives Dw14.2 (Gly56) and Dw14.1 (Val ¶*-which differ only at this one position in the entire antigen-binding region-show an all-or-nothing difference in presenting activity. This dimorphism at position 86 is widespread, occurring in subtypes of DR1, DR2, DR3, DR5, and DR6 alleles as well as DR4. Since other DR4 subtypes with substitutions at positions 70-74 also fail to present this peptide, and glycine residues can be uniquely flexible, we suggest that this replacement at position 86 acts locally or at a distance by altering the conformation of the peptide-binding cleft. Such profound functional consequences for T-cell recognition as we report here may explain this example of conserved major histocompatibility complex diversity.The striking polymorphism of the major histocompatibility complex (MHC) in most vertebrate species implies strong selection for diversity. As functional and crystallographic studies show (for class I molecules), the role of these MHC heterodimers is to bind peptide fragments of processed antigens in a cleft formed by the folding of their two membrane-distal domains (1, 2) and to present them to T lymphocytes. [For CD4' helper T cells, the class II molecules on specialized antigen-presenting cells (APCs) fulfill this role.] Specific recognition of the MHC and peptide components of the resulting composite by T lymphocytes underlies the phenomenon of MHC restriction whereby most peptides are recognized in the context of only some of the class II alleles. Thus, the diversity of the MHC molecules probably ensures that the population as a whole maintains responsiveness to a broad repertoire of foreign antigens.Many autoimmune diseases show associations with particular MHC alleles or haplotypes (reviewed in ref.3).

show abstract

“…The value of using transfectants as tools for the fine mapping of monoclonal antibodies is also illustrated in Table 3 (Altmann et al 1990). The antibody CA2 or 2.06 (Charron and McDevitt 1979), which is apparently monomorphic in its reactions with lymphoid lines, can be seen not to react with the DR 7 antigen, when the latter is presented on a transfectant in isolation from the other class II region products.…”

Section: Transfectantsmentioning

confidence: 98%

“…For this reason transfectants are very useful. Transfectants, such as those listed in Table 2, are mouse cells (Altmann et al 1990) or class II-negative human B cells (Merkenschlager et al 1991) into which genomic or cDNA clones have been introduced in such a way as to allow the product of those particular genes to be expressed on the cell surface. Followjng transfection the cells which express the highest level of antigen, measured by their ability to bind antibody, are selected and cloned (Sambrook et al 1989).…”

Section: Transfectantsmentioning

confidence: 99%

B Cell Recognition and HLA Typing: Current Methods and Future Possibilities. Role of Alloantibodies and Monoclonal Antibodies as Reagents

Bodmer¹,

Marsh²,

Heyes³

et al. 1993

The HLA System in Clinical Transplantation

View full text Add to dashboard Cite

Fine mapping of HLA class II monoclonal antibody specificities using transfected L cells

Cited by 27 publications

References 25 publications

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

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

Critical role for the Val/Gly86 HLA-DR beta dimorphism in autoantigen presentation to human T cells.

B Cell Recognition and HLA Typing: Current Methods and Future Possibilities. Role of Alloantibodies and Monoclonal Antibodies as Reagents

Contact Info

Product

Resources

About