TY - JOUR
T1 - Granzyme B-dependent and perforin-independent DNA fragmentation in intestinal epithelial cells induced by anti-CD3 mAb-activated intra-epithelial lymphocytes
AU - Ogata, Masaki
AU - Ota, Yuta
AU - Matsutani, Takaji
AU - Nanno, Masanobu
AU - Suzuki, Ryuji
AU - Itoh, Tsunetoshi
N1 - Funding Information:
This work was in part supported by a Grant-in-aid for Scientific Research from the Ministry of Education, Science and Culture, Japan (20590181 to M.O. and 21590207 to T.I.). M.Ogata(*).Y.Ota.T.Itoh Division of Immunology and Embryology, Department of Cell Biology, Tohoku University School of Medicine, 980-8575 Sendai, Japan e-mail: ogata@immem.med.tohoku.ac.jp
PY - 2013/5
Y1 - 2013/5
N2 - We previously found that an i.p. injection of anti-CD3 monoclonal antibody (mAb) into mice caused DNA fragmentation in the intestinal villous epithelial cells (IVECs) of the duodenum and the jejunum. In this study, in order to elucidate the mechanism of DNA fragmentation in IVECs, we searched for the inducer(s) of DNA fragmentation by using immunohistochemistry. The release of cytoplasmic granules from intraepithelial lymphocytes (IELs) and the formation of large gaps between IELs and IVECs were observed electron microscopically after antibody administration. The presence and distribution pattern of Granzyme B (GrB), a serine protease in cytolytic granules present in cytotoxic T lymphocytes and natural killer cells and considered to be the responsible molecule for DNA fragmentation in target cells, was examined in detail in intestinal villi by immunohistology. GrB was detected in cytoplasmic granules in nearly all IELs. The time-kinetics of granule release from IELs after mAb injection coincided not only with that of the extracellular diffusion of GrB, but also with that of DNA fragmentation in IVECs. On the other hand, perforin (Pfn), assumed to cooperate with GrB in DNA fragmentation, could not be detected in IELs, and its release was not confirmed after the anti-CD3 mAb injection. Anti-CD3 mAb injection also induced DNA fragmentation in IVECs in Pfn-knockout mice. These results support the notion that DNA fragmentation in IVECs by the stimulated IELs in the present study is induced by a mechanism involving GrB, but independent of Pfn.
AB - We previously found that an i.p. injection of anti-CD3 monoclonal antibody (mAb) into mice caused DNA fragmentation in the intestinal villous epithelial cells (IVECs) of the duodenum and the jejunum. In this study, in order to elucidate the mechanism of DNA fragmentation in IVECs, we searched for the inducer(s) of DNA fragmentation by using immunohistochemistry. The release of cytoplasmic granules from intraepithelial lymphocytes (IELs) and the formation of large gaps between IELs and IVECs were observed electron microscopically after antibody administration. The presence and distribution pattern of Granzyme B (GrB), a serine protease in cytolytic granules present in cytotoxic T lymphocytes and natural killer cells and considered to be the responsible molecule for DNA fragmentation in target cells, was examined in detail in intestinal villi by immunohistology. GrB was detected in cytoplasmic granules in nearly all IELs. The time-kinetics of granule release from IELs after mAb injection coincided not only with that of the extracellular diffusion of GrB, but also with that of DNA fragmentation in IVECs. On the other hand, perforin (Pfn), assumed to cooperate with GrB in DNA fragmentation, could not be detected in IELs, and its release was not confirmed after the anti-CD3 mAb injection. Anti-CD3 mAb injection also induced DNA fragmentation in IVECs in Pfn-knockout mice. These results support the notion that DNA fragmentation in IVECs by the stimulated IELs in the present study is induced by a mechanism involving GrB, but independent of Pfn.
KW - DNA fragmentation
KW - Granzyme B
KW - Intraepithelial lymphocytes
KW - Mouse (female, C57BL/6)
KW - Perforin
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U2 - 10.1007/s00441-012-1549-7
DO - 10.1007/s00441-012-1549-7
M3 - Article
C2 - 23361111
AN - SCOPUS:84880570050
VL - 352
SP - 287
EP - 300
JO - Zeitschrift für Zellforschung und mikroskopische Anatomie (Vienna, Austria : 1948)
JF - Zeitschrift für Zellforschung und mikroskopische Anatomie (Vienna, Austria : 1948)
SN - 0302-766X
IS - 2
ER -