Identification of HLA-DR-bound peptides presented by human bronchoalveolar lavage cells in sarcoidosis.
Sarcoidosis is an inflammatory disease of unknown etiology, most commonly affecting the lungs. Activated CD4+ T cells accumulate in the lungs of individuals with sarcoidosis and are considered to be of central importance for inflammation. We have previously shown that Scandinavian sarcoidosis patients expressing the HLA-DR allele DRB1*0301 are characterized by large accumulations in the lungs of CD4+ T cells expressing the TCR AV2S3 gene segment. This association afforded us a unique opportunity to identify a sarcoidosis-specific antigen recognized by AV2S3+ T cells. To identify candidates for the postulated sarcoidosis-specific antigen, lung cells from 16 HLA-DRB1*0301pos patients were obtained by bronchoalveolar lavage. HLA-DR molecules were affinity purified and bound peptides acid eluted. Subsequently, peptides were separated by reversed-phase HPLC and analyzed by liquid chromatography-mass spectrometry. We identified 78 amino acid sequences from self proteins presented in the lungs of sarcoidosis patients, some of which were well-known autoantigens such as vimentin and ATP synthase. For the first time, to our knowledge, we have identified HLA-bound peptides presented in vivo during an inflammatory condition. This approach can be extended to characterize HLA-bound peptides in various autoimmune settings.
Wahlström
Jan
J
Lung Research Laboratory, Department of Medicine, Karolinska Institutet, Stockholm, Sweden. jan.wahlstrom@ki.se
Dengjel
Jörn
J
Persson
Bengt
B
Duyar
Hüseyin
H
Rammensee
Hans-Georg
HG
Stevanović
Stefan
S
Eklund
Anders
A
Weissert
Robert
R
Grunewald
Johan
J
D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology*; D005260:Female; D006684:HLA-DR Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D059811:HLA-DRB1 Chains; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017565:Sarcoidosis, Pulmonary / Q000235:genetics / Q000276:immunology* / Q000473:pathology
D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; C553502:HLA-DRB1*03:01 antigen; D010455:Peptides