| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| Research Abstract |
1.The aim of the study is to investigate the devastation process of human renal grafts through rejection by focusing on Fas antigen which causes apoptosis.
2.The plan and method : Materials consisted of 2 one hour biopsies, 26 biopsies, 26 biopsies taken at posttrasplant 9 days to 7 years and 9 months, and 2 removed rejected grafts. As a control, a non-cancer portion of the kidney with carcinoma was used. Materials were frozen or formalin-fixed and paraffin-embedded. Methods were (1) immunostaining for Fas, DR,and bcl-2 proteins, (2) immunostaining for apoptotic DNA,(3) in-situ-hybridization (ISH) of mRNA of Fas and IFNgamma, and (4) light and electron microscopical observation.
3.Results : In a control kidney the expression of Fas, DR,and bcl-2 as well as apoptosis was not observed. However, nuclei clearly stained with ApopTag kit were spottedly present in the straight portion of proximal convoluted tubules (prox.conv.tub.). The results of renal grafts investigated were as follows. The
… More
expression of Fas was relatively strong in prox.conv.tub.and collecting tub.. No expression was observed in the endothelium of vessels. Infiltrating cells were generally negative for it but weakly positive in few materials. The expression of bcl-2 was only recognized in prox.conv.tub..Most infiltrating cells were intensely positive. The expression of DR was similar to that of Fas in tub. but somewhat weaker. The expression of vessel endothelia was strong in capillaries and weak in large arteries. The most part of infiltrating cells expressed it strongly. Apoptotic DNA clearly stained was only recognized in the nuclei of detached epithelia in tubular lumens. Otherwise weakly stained nuclei were scattered. Light and electron microscopical observations : apoptosis was not observed. ISH of mRNA of Fas and IFNgamma was not successful. Reinvestigation is in progress by use of an RNA probe.
4.Discussion : That cells simultancously expressed both Fas and bcl-2 seems to suggest that apoptosis does not so easily occur. Actually, it was not observed light and electron microscopically and immunohistochemically. It is noteworthy that any endothelia which formed the fundamental structure of tissues did not express Fas antigen. That the endothelium is resistnt to apoptosis is favorable for life. Less
|
