title: Mass spectrometric characterization of circulating and functional antigens derived from piperacillin in patients with cystic fibrosis.
abstract: A mechanistic understanding of the relationship between the chemistry of drug Ag formation and immune function is lacking. Thus, mass spectrometric methods were employed to detect and fully characterize circulating Ags derived from piperacillin in patients undergoing therapy and the nature of the drug-derived epitopes on protein that can function as an Ag to stimulate T cells. Albumin modification with piperacillin in vitro resulted in the formation of two distinct haptens, one formed directly from piperacillin and a second in which the dioxopiperazine ring had undergone hydrolysis. Modification was time and concentration dependent, with selective modification of Lys(541) observed at low concentrations, whereas at higher concentrations, up to 13 out of 59 lysine residues were modified, four of which (Lys(190), Lys(195), Lys(432), and Lys(541)) were detected in patients' plasma. Piperacillin-specific T lymphocyte responses (proliferation, cytokines, and granzyme B release) were detected ex vivo with cells from hypersensitive patients, and analysis of incubation medium showed that modification of the same lysine residues in albumin occurred in situ. The antigenicity of piperacillin-modified albumin was confirmed by stimulation of T cells with characterized synthetic conjugates. Analysis of minimally modified T cell-stimulatory albumin conjugates revealed peptide sequences incorporating Lys(190), Lys(432), and Lys(541) as principal functional epitopes for T cells. This study has characterized the multiple haptenic structures on albumin in patients and showed that they constitute functional antigenic determinants for T cells.
authors: Regional Adult Cystic Fibrosis Unit, St. James's Hospital, Leeds LS9 7TF, United Kingdom.
authors: Paul
authors: P
authors: Whitaker
authors: Xiaoli
authors: X
authors: Meng
authors: Sidonie N
authors: SN
authors: Lavergne
authors: Sabah
authors: S
authors: El-Ghaiesh
authors: Manal
authors: M
authors: Monshi
authors: Caroline
authors: C
authors: Earnshaw
authors: Daniel
authors: D
authors: Peckham
authors: Jimmy
authors: J
authors: Gooi
authors: Steve
authors: S
authors: Conway
authors: Munir
authors: M
authors: Pirmohamed
authors: Rosalind E
authors: RE
authors: Jenkins
authors: Dean J
authors: DJ
authors: Naisbitt
authors: B Kevin
authors: BK
authors: Park
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