Recombinant HLA-DP2 binds beryllium and tolerizes beryllium-specific pathogenic CD4+ T cells.
Chronic beryllium disease is a lung disorder caused by beryllium exposure in the workplace and is characterized by granulomatous inflammation and the accumulation of beryllium-specific, HLA-DP2-restricted CD4+ T lymphocytes in the lung that proliferate and secrete Th1-type cytokines. To characterize the interaction among HLA-DP2, beryllium, and CD4+ T cells, we constructed rHLA-DP2 and rHLA-DP4 molecules consisting of the alpha-1 and beta-1 domains of the HLA-DP molecules genetically linked into single polypeptide chains. Peptide binding to rHLA-DP2 and rHLA-DP4 was consistent with previously published peptide-binding motifs for these MHC class II molecules, with peptide binding dominated by aromatic residues in the P1 pocket. 9Be nuclear magnetic resonance spectroscopy showed that beryllium binds to the HLA-DP2-derived molecule, with no binding to the HLA-DP4 molecule that differs from DP2 by four amino acid residues. Using beryllium-specific CD4+ T cell lines derived from the lungs of chronic beryllium disease patients, beryllium presentation to those cells was independent of Ag processing because fixed APCs were capable of presenting BeSO4 and inducing T cell proliferation. Exposure of beryllium-specific CD4+ T cells to BeSO4 -pulsed, plate-bound rHLA-DP2 molecules induced IFN-gamma secretion. In addition, pretreatment of beryllium-specific CD4+ T cells with BeSO4-pulsed, plate-bound HLA-DP2 blocked proliferation and IL-2 secretion upon re-exposure to beryllium presented by APCs. Thus, the rHLA-DP2 molecules described herein provide a template for engineering variants that retain the ability to tolerize pathogenic CD4+ T cells, but do so in the absence of the beryllium Ag.
Fontenot
Andrew P
AP
Departments of Medicine and Immunology, University of Colorado Health Sciences Center, Denver, CO 80206, USA.
Keizer
Timothy S
TS
McCleskey
Mark
M
Mack
Douglas G
DG
Meza-Romero
Roberto
R
Huan
Jianya
J
Edwards
David M
DM
Chou
Yuan K
YK
Vandenbark
Arthur A
AA
Scott
Brian
B
Burrows
Gregory G
GG
D000595:Amino Acid Sequence; D000818:Animals; D001607:Berylliosis / Q000276:immunology* / Q000628:therapy; D001608:Beryllium / Q000276:immunology / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D002460:Cell Line; D002908:Chronic Disease; D006682:HLA-DP Antigens / Q000235:genetics / Q000378:metabolism*; D059868:HLA-DP beta-Chains; D006801:Humans; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
D006682:HLA-DP Antigens; D059868:HLA-DP beta-Chains; C083176:HLA-DPw2 antigen; D008024:Ligands; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; D001608:Beryllium