본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Kidney international v.49 no.2, 1996년, pp.403 - 412  

Apoptosis of cultured rat glomerular mesangial cells induced by IgG2a monoclonal anti-Thy-1 antibodies.

Sato, T ; van Dixhoorn, M G ; Schroeijers, W E ; Huizinga, T W ; Reutelingsperger, C P ; van Es, L A ; Daha, M R ;
  • 초록  

    Anti-Thy-1 nephritis is a model of mesangial proliferative glomerulonephritis. It has been suggested that apoptosis, which is a counteracting regulatory mechanism against undesired cell proliferation, is involved in sequential histological changes in this model. In the present study, we investigated whether IgG2a anti-Thy-1 monoclonal antibody (ER4) or its F(ab')2 fragments are able to induce apoptosis of rat glomerular mesangial cells (GMC) in vitro. After co-culture with ER4 or its F(ab')2 fragments, apoptosis was assessed by morphological studies with Hoechst 33258 stain and FITC-annexin V. The latter detects the dislocation of negatively charged phospholipid, phosphatidylserine, from the inner to the outer leaflet of the membrane during apoptosis. This is a sensitive method for the detection of apoptosis. Under fluorescent microscopy, distinct nuclear condensation and positive reactivity with FITC-annexin V were observed in cells co-cultured with ER4 or its F(ab')2 fragments. The results obtained by FACS analysis with annexin V showed a direct correlation with the detection of apoptosis with the terminal deoxynucleotidyl transferase reaction (TDT). Up to 19% and 23% of rat GMC, which were co-cultured for 24 hours with 1 microgram/ml (0.5 microgram/l x 10(5) cells) of ER4 or its F(ab')2 fragments, were labeled by TDT, respectively. With annexin V, up to 34% and 31% cells displaying apoptosis were seen. The degree of apoptosis as measured by the annexin V method was dependent on the concentration of ER4 and time of incubation in the presence of ER4. Finally, apoptosis was confirmed by gel electrophoresis of DNA isolated from the cells co-cultured with each monoclonal antibody (MAb). DNA extracts from cells co-cultured with ER4 or its F(ab')2 fragments demonstrated typical internucleosomal DNA fragmentation. Medium alone, controls of anti-human C3bi receptor MAb (IB4) and anti-rat MHC class I MAb (OX18) showed neither nuclear changes nor significant labeling of the cells with the TDT reaction or with the annexin V. Taken together, these results demonstrate for the first time that anti-Thy-1 MAb is able to induce apoptosis of rat GMC in vitro. The Thy-1 antigen on rat GMC, therefore, seems to function as one of the molecules regulating cell death and thereby may determine the degree of mesangial alteration in Thy-1 nephritis.


  • 원문보기

    원문보기
    무료다운로드 유료다운로드
    • 유료 원문 정보가 존재하지 않습니다.

    유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

    NDSL에서는 해당 원문을 복사서비스하고 있습니다. 아래의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역