Motif of HLA-B*3503 peptide ligands

✍ Scribed by Alexander Steinle; Kirsten Falk; Olaf Rötzschke; Volker Gnau; Stefan Stevanović; Günther Jung; Dolores J. Schendel; Hans-Georg Rammensee

• Publisher: Springer-Verlag
• Year: 1995
• Tongue: English
• Weight: 253 KB
• Volume: 43
• Category: Article
• ISSN: 0093-7711
• DOI: 10.1007/bf00186615

✦ Synopsis

HLA-B*3501 and HLA-B*3503 molecules differ by a single amino acid located in the [~-pleated sheet of the c~2 domain. In order to clarify the basis of the strict distinction between these highly related class I molecules by several alloreactive T cell clones , we determined the motif of HLA-B*3503 peptide ligands and compared it with the previously published HLA-B*3501 motif .

HLA-B*3503 full-length cDNA (1.35 kilobases) was amplified by HLA-B locus-specific polymerase chain reaction from cDNA of donor HF and cloned in pUC18 as described . A fully sequenced cDNA clone devoid of Taq-DNA pol, ymerase incorporation errors was cloned into the eucaryotic expression vector pFM92.1 (kindly provided by G. H~immerling, Heidelberg), a derivative of pHI3APr-1 neo . Cotransfection with the human gene for ~2-microglobulin of the mouse mastocytoma cell line P815 was performed using Lipofectin (Gibco BRL) according to manufacturer's instructions. Staining with the HLA-B-specific antibody 4E ) revealed good surface expression of HLA-B*3503 on G418-resistant P815 cells (data not shown). These were expanded to obtain about 30 ml of cell pellet, lysed and HLA-B*3503 mole-