| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Outline of Annual Research Achievements |
Anti-laminin (LM) γ1 pemphigoid is an autoimmune blistering disease with IgG antibodies to LMγ1. To characterize anti-LMγ1 autoantibodies, sera from 24 anti-LMγ1 pemphigoid patients and 6 normal controls were harvested. Normal human dermal extract (NHDE), five different cell lines, and recombinant proteins (RPs) of LMγ1 were employed as LMγ1 antigen sources for immunoblotting (IB) and immunofluorescence (IF). Unexpectedly, the molecular weights (MWs) of LMγ1 in lysates of five cell lines are different. After deglycosylation with PNGase F, the MWs of LMγ1 become smaller and identical. Patient sera failed to react with LMγ1 in five cell lysates or culture supernatants by IB and showed negative results in IF of cell lines, although all patient sera can react with LMγ1 in NHDE. In contrast, 20 (83%) and 18 (75%) of 24 patient sera reacted with LMγ1 in LM521 and LM411 RPs, respectively. In addition, patient sera failed to react with LMγ1 in deglycosylated cell lysates or supernatants. Compared to those without deglycosylation, patient sera reacted more weakly with deglycosylated LMγ1 in NHDE and more strongly with deglycosylated LMγ1 in LM521, LM411 and C-terminal domain of LM511 RPs. In summary, anti-LMγ1 antibodies in patient sera are heterogeneous. N-linked glycosylation influences at least partly the reactivity of anti- LMγ1 antibodies. This work has been published recently (Li et al. J Dermatol Sci. 2015 77(2):125-9.)
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