RT Journal Article
SR Electronic
T1 A mechanism for acceleration of GM-CSF autoantibody (GMAb) production in autoimmune pulmonary alveolar proteinosis (aPAP)
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP P2024
VO 44
IS Suppl 58
A1 Koh Nakata
A1 Takahiro Tanaka
A1 Ryushi Tazawa
YR 2014
UL http://erj.ersjournals.com/content/44/Suppl_58/P2024.abstract
AB Background:IgG type GMAb is the etiologic agent for aPAP, although low concentration of the antibody is present in healthy subjects. To elucidate whether the acceleration of IgG-GMAb production by B cells in aPAP is caused by expansion/maturation of the corresponding B cells, we evaluated the frequencies and sequence of IgG-BCR+ B cell clones in aPAP and normal subjects (control). Methods: Memory B cells were sorted using anti-CD27 magnetic beads from blood cells,inoculated with Epstein Barr virus (EBV), and GMAb producing B cells were detected by ELISPOT and ELISA.GMAb BCR+ were sorted magnetically followed by RNA extraction and cDNA generation. IgH cDNA V region were amplified by PCR and were applied to the next generation sequencing (GS FLX and GS Junior Systems, Roche®). The sequence data were analyzed using a bioinformatics software. Results: ELISA and ELISPOT data demonstrated that GMAb producing cells was less frequently in the controls than aPAP. The next generation sequencing of membrane-GMAb positice B cells revealed that the number of productive IgG sequences was much larger in aPAP than the controls. CDR2 sequences tended to form cluster by phylogenic analysis especially in severe aPAP, suggesting that clonal expansion and selection may proceed in the disease process. Thus, our present data suggest that the accelerated IgG-GMAb BCR+ B cell clones are expanding in aPAP.