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標題: | 犬CD5lo細胞誘發CD5-CD21-細胞的自然殺手細胞
之細胞毒殺能力 Induction of NK cytotoxicity of canine CD5/CD21 double negative cells by CD5lo NK cells |
作者: | Chun-Pei Chang 張君珮 |
指導教授: | 朱瑞民 |
共同指導教授: | 廖泰慶 |
關鍵字: | 犬自然殺手細胞, canine natural killer cells,peripheral blood lymphocytes (PBLs),lymphokine-activated killers (LAKs),CD5lo cells,CD5- cells,non-T non-B,natural cytotoxicity, |
出版年 : | 2011 |
學位: | 碩士 |
摘要: | 自然殺手細胞(NK細胞)對於免疫系統之調控,以及在病毒與腫瘤之對抗中均扮演著極為重要的角色。然而目前的研究對於犬的自然殺手細胞卻瞭解不多。我們先前的發表首度發現犬周邊血液淋巴球中CD5低表現量(CD5lo)細胞,具有明顯自然殺手細胞之特性,且被認為是犬NK細胞內非常重要的族群。此族群有別於其他已發現之哺乳類動物的NK細胞,因其具有T細胞特徵性之分子標記CD3、CD8和α/βTCR,此一發現使得犬隻NK細胞在比較免疫學中成為極為特殊的存在。本文針對犬NK細胞的深入研究則顯示,原本僅具微弱毒殺能力且表現型為CD21-與CD5lo-ne之細胞群,在與CD5lo細胞共同培養後,竟會產生高度NK細胞專一性的細胞毒殺能力。而該現象僅限於CD5lo細胞,其他如CD5hi、CD21+和嗜中性細胞皆無法增加其毒殺能力。以高階細胞流式分選儀將CD5lo-ne細胞區分成CD5- 和CD5hi兩族群進一步作測試,結果顯示CD5-細胞在去除CD5lo細胞單獨培養之情況下,不論是否經過IL-2刺激其細胞毒殺能力都偏低(當E/T ration 50:1,毒殺能力為4.16±3.51%)。相較於兩者共同培養時,CD5-細胞的NK細胞毒殺能力則明顯被誘發(當E/T ration 50:1,毒殺能力為22.42±18.07%)。而CD5lo-ne細胞的另一組成者CD5hi細胞,以同樣的實驗方法,縱使有CD5lo細胞存在也不具有細胞毒殺能力(當E/T ration 50:1,毒殺能力為3.60±2.92% or 3.07±2.08%)。使用更精細的細胞分選並進行NK細胞毒殺能力之測試結果顯示,CD5-細胞中的CD5-CD21-細胞是在CD5lo細胞誘發下,主要具有NK細胞毒殺力的細胞。這是第一篇報告證明,NK細胞可誘發其他具自然殺手細胞潛質之細胞族群表現出NK細胞毒殺能力。在米格魯犬中,除了先前已發現的CD5lo細胞外,CD5-CD21-細胞在經由CD5lo細胞之誘發下亦能產生NK細胞活性。 Natural killer (NK) cells play important roles in both early protection against viruses or tumor cells and in regulating immune functions. However, only a few studies have focused on the characterization of canine NK cells. We previously demonstrated that canine peripheral blood lymphocytes with low surface CD5 density (CD5lo) are closely associated with NK cell characteristics, and are considered to be an important NK population. NK cells in most mammals do not express T cell markers. However, CD5lo cells expressed surface molecules such as CD3, CD8, and α/βTCR, making canines a unique model to study comparative immunology in NK cells. In this study, we discovered that although CD5lo (CD5lo-ne) and CD21 double negative cells were originally low in NK cytotoxicity, they became highly activated in NK cytotoxicity when cultured with CD5lo NK cells. The cytotoxicity was not activated as in the culture of other types of cells, such as CD5hi cells and CD21+. The CD5lo-ne population comprises CD5- and CD5hi cells, which were sorted separately by fluorescent density with a FACSAria TM flow cytometer. We first tested CD5- cells and found that these cells were initially low in NK cytotoxicity or after being cultured in IL-2 when CD5lo cells were absent in the culture (4.16±3.51% at ET ratio of 50:1), but as CD5lo cells were added in similar medium, the NK activity became greatly enhanced (22.42±18.07at ET ratio of 50:1). The other population of CD5lo-ne, CD5hi cells, was always NK inactive, no matter whether or not it was cultured alone or with CD5lo cells (3.60±2.92% or 3.07±2.08% at ET ratio of 50:1). We further verified that CD5- cells carrying no CD21 surface molecules, i.e. CD5/CD21 double negative cells were the cell type in CD5- cells population whose NK cytotoxicity was activated by CD5lo cells. This is the first report to present that NK cells enhanced NK-inert cells to become NK cytotoxic. It also concluded that in beagles, besides CD5lo cells, CD5/CD21 double negative cells also exhibited NK activity. However, it depended on the presence of CD5lo cells |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/37584 |
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