Study of Activation-induced Cell Death of Natural Killer Cell

碩士 === 台北醫學院 === 細胞及分子生物研究所 === 85 === Human natural killer cells (NK) play important roles in innate immune system and have the potential to regulate the adapted immune responses. During a immune response, most activated lymphocytes eventu...

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Main Authors: Lee, Jann-Mou, 李建謀
Other Authors: Nan-Shih Liao
Format: Others
Language:zh-TW
Published: 1997
Online Access:http://ndltd.ncl.edu.tw/handle/20139873068368623640
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spelling ndltd-TW-085TMC003390012016-07-01T04:15:58Z http://ndltd.ncl.edu.tw/handle/20139873068368623640 Study of Activation-induced Cell Death of Natural Killer Cell 自然殺手細胞活化致死之研究 Lee, Jann-Mou 李建謀 碩士 台北醫學院 細胞及分子生物研究所 85 Human natural killer cells (NK) play important roles in innate immune system and have the potential to regulate the adapted immune responses. During a immune response, most activated lymphocytes eventually died. Especially, restimulating activated T cells by triggering CD3/TCR complex would make them undergo apoptosis, a process termed as activation-induced cell death: AICD. Thus, we intend to determine the occurrence of AICD on NK cells and mechanisms directing the AICD. First of all, we stimulated cells by the treatment of PMA/Ionophore or crosslinking CD16 or Kp43 molecules on the surface of NK cells ,and found that by the treatment of PMA /Ionophore or crosslinking CD16 molecules for 12-15 hours, 60-70% of short- term cultured NK cells died in the form of apoptosis but not for the crosslinking of Kp43 molecules. We further determine the roles of Fas and LTbR, two members of TNF receptor super-family in the AICD of NK cells. Through analysis toward surface molecules on NK cells by Flow Cytometer, we have found that freshly isolated NK cells could express low level of Fas molecules, pretty high amounts of LTb and LTbR molecules but no Fas ligand. Two-week in-vitro culture later, the only difference is the enrichment of Fas molecules. By the treatment of PMA /Ionophore or crosslinking CD16 molecules, Fas ligand molecules were induced, Fas molecules were with same expressive pattern but the expressive amounts of LTb and LTbR molecules were significantly reduced to 30-50% of the original ones. Stimulation upon crosslinking Kp43 molecules have no significant effects on the expressive patterns of the four molecules. Then we took advantage of Baculovirus protein expression system to synthesize soluble Fas-Fc and soluble LTbR-Fc proteins and found that s-Fas-Fc could prevent cells from apoptosis which resulted from stimulation of PMA/Ionophore or cross- linking CD16 ,but s- LTbR-Fc could not. Finally, the usage of IL-4 has been shown to block the AICD of NK cells in a dose-dependent manner. In summary, we proved that NK cells could undergo " AICD" upon crosslinking the activation-receptor, CD16 and Fas mediated apoptosis played the major roles in regulating AICD, but not for LTbR mediated apoptosis. In contrast, signals delivered from LTbR would inhibit apoptosis and therefor the interactions betweem LTb and LTbR were essential for survival of human NK cells. Nan-Shih Liao 廖南詩 1997 學位論文 ; thesis 62 zh-TW
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description 碩士 === 台北醫學院 === 細胞及分子生物研究所 === 85 === Human natural killer cells (NK) play important roles in innate immune system and have the potential to regulate the adapted immune responses. During a immune response, most activated lymphocytes eventually died. Especially, restimulating activated T cells by triggering CD3/TCR complex would make them undergo apoptosis, a process termed as activation-induced cell death: AICD. Thus, we intend to determine the occurrence of AICD on NK cells and mechanisms directing the AICD. First of all, we stimulated cells by the treatment of PMA/Ionophore or crosslinking CD16 or Kp43 molecules on the surface of NK cells ,and found that by the treatment of PMA /Ionophore or crosslinking CD16 molecules for 12-15 hours, 60-70% of short- term cultured NK cells died in the form of apoptosis but not for the crosslinking of Kp43 molecules. We further determine the roles of Fas and LTbR, two members of TNF receptor super-family in the AICD of NK cells. Through analysis toward surface molecules on NK cells by Flow Cytometer, we have found that freshly isolated NK cells could express low level of Fas molecules, pretty high amounts of LTb and LTbR molecules but no Fas ligand. Two-week in-vitro culture later, the only difference is the enrichment of Fas molecules. By the treatment of PMA /Ionophore or crosslinking CD16 molecules, Fas ligand molecules were induced, Fas molecules were with same expressive pattern but the expressive amounts of LTb and LTbR molecules were significantly reduced to 30-50% of the original ones. Stimulation upon crosslinking Kp43 molecules have no significant effects on the expressive patterns of the four molecules. Then we took advantage of Baculovirus protein expression system to synthesize soluble Fas-Fc and soluble LTbR-Fc proteins and found that s-Fas-Fc could prevent cells from apoptosis which resulted from stimulation of PMA/Ionophore or cross- linking CD16 ,but s- LTbR-Fc could not. Finally, the usage of IL-4 has been shown to block the AICD of NK cells in a dose-dependent manner. In summary, we proved that NK cells could undergo " AICD" upon crosslinking the activation-receptor, CD16 and Fas mediated apoptosis played the major roles in regulating AICD, but not for LTbR mediated apoptosis. In contrast, signals delivered from LTbR would inhibit apoptosis and therefor the interactions betweem LTb and LTbR were essential for survival of human NK cells.
author2 Nan-Shih Liao
author_facet Nan-Shih Liao
Lee, Jann-Mou
李建謀
author Lee, Jann-Mou
李建謀
spellingShingle Lee, Jann-Mou
李建謀
Study of Activation-induced Cell Death of Natural Killer Cell
author_sort Lee, Jann-Mou
title Study of Activation-induced Cell Death of Natural Killer Cell
title_short Study of Activation-induced Cell Death of Natural Killer Cell
title_full Study of Activation-induced Cell Death of Natural Killer Cell
title_fullStr Study of Activation-induced Cell Death of Natural Killer Cell
title_full_unstemmed Study of Activation-induced Cell Death of Natural Killer Cell
title_sort study of activation-induced cell death of natural killer cell
publishDate 1997
url http://ndltd.ncl.edu.tw/handle/20139873068368623640
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