Evaluation of molecular mechanisms of dialysis related amyloidosis and the diagnosis method of early detection

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New monoclonal antibody to c-terminal of β 2-microglobulin (β 2MG) was prepared and investigated the importance of pathogeneses of dialysis related amyloidosis (DRA) or end stage renal disease.We characterized new monoclonal antibody to c-terminal of β 2MG 92-99. β2MG extracted from ultrafiltrate showed no reaction form mAb β 2MG 92-99, whereas acidified b2mfrom ultrafiltrate showed a little reaction for form mAb β2MG 92-99. A homogenate of carpal amyloid tissues showed a strong reaction for mAb β2MG 92-99 on immunoblotting. We considered that mAb β2MG 92-99 didn't react native form of β2MG, but unfolded and change of structure β2MG, mAb β 2MG 92-99 react unfolded β2MG. In the monocyte cell line, temperature stress, osmotic pressure stress, radical stress unregulated the cell surface unfolded β2MG expression. The same stresses also unregulated the GRP78 mRNA expression.In the maintenance hemodialysis patients, cell surface unfolded β2MG expression was increased compare to normal subjects. Moreover, GRP78 mRNA expression was also increased in maintenance hemodialysis patients, compared to normal subjects.Immunohistochemical study showed also a distinct staining for mAb 92-99 in congophilic specimens from DRA patients more interestingly, staining form mAb β2MG 92-99 could be found in most, though not all, non-congophilic tissues. The monoclonal antibody specific to the C-terminal 92-99 of β2MG 92-99 can detect the conformational intermediate in amyloidogenesis of b2m ex vivo, and demonstrates that an unfolded β2MG at C-terminal could be found not only in congophilic area but even in non-congophilic area as well.