三氯乙烯藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究

1、目目 錄錄 中文摘要中文摘要.1 1 英文摘要英文摘要.6 6 英文縮寫詞表英文縮寫詞表.1212 前前 言言.1414 第一部分第一部分 TCETCE 藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究 .1919 第一節(jié) TCE 藥疹樣皮炎患者外周血淋巴細(xì)胞抗原特異性增殖功能測(cè) 定.2121 第二節(jié) TCE 藥疹樣皮炎患者外周血免疫細(xì)胞檢測(cè)分析 .2626 第三節(jié) Fas/FasL 與 PF/GrB 介導(dǎo)的 TCE 免疫毒作用機(jī)制探討 .3737 小 結(jié).5555 第二部分第二部分 8 8 例例 TCETCE 藥疹樣皮炎患者病例報(bào)告分析藥疹樣皮炎患者病例報(bào)告分析

2、.5656 小 結(jié).7171 總總 結(jié)結(jié).7272 致致 謝謝.7474 參考文獻(xiàn)參考文獻(xiàn).7676 附錄附錄 1 1 生物標(biāo)志物的驗(yàn)證及其在健康危險(xiǎn)預(yù)警中的作用生物標(biāo)志物的驗(yàn)證及其在健康危險(xiǎn)預(yù)警中的作用 .8686 附錄附錄 2 2 個(gè)人簡(jiǎn)歷個(gè)人簡(jiǎn)歷 .9696 三氯乙烯藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究三氯乙烯藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究 中文摘要中文摘要 職業(yè)接觸三氯乙烯（Trichloroethylene，TCE）引起的 TCE 藥疹樣皮炎是 近年來我國職業(yè)衛(wèi)生領(lǐng)域令人關(guān)注的問題，但目前關(guān)于該病的發(fā)病機(jī)制尚不清 楚，對(duì) TCE 職業(yè)接觸工人缺乏特異性健康監(jiān)護(hù)指標(biāo)，對(duì) TC

3、E 藥疹樣皮炎患者缺 乏特異性臨床治療措施。 根據(jù)該病的發(fā)病特點(diǎn)一般認(rèn)為其屬于型超敏反應(yīng)的可能性最大，型超 敏反應(yīng)發(fā)生的機(jī)制與抗體和補(bǔ)體無關(guān)，主要是 T 細(xì)胞介導(dǎo)的免疫損傷。T 細(xì)胞 介導(dǎo)的免疫反應(yīng)發(fā)生的過程涉及到記憶性 T 淋巴細(xì)胞的生成和效應(yīng)性 T 細(xì)胞的 活化增殖，效應(yīng)性 T 細(xì)胞通過分泌一系列細(xì)胞因子發(fā)揮作用，并調(diào)節(jié) B 細(xì)胞、 NK 細(xì)胞以及其他各群免疫細(xì)胞的活化增殖。另外，細(xì)胞毒性 T 細(xì)胞（CTL）在 T 細(xì)胞介導(dǎo)的超敏反應(yīng)中發(fā)揮重要的作用，其主要通過兩條途徑殺傷靶細(xì)胞， 其一是穿孔素-顆粒酶途徑，其主要參與免疫防御；其二是 Fas-FasL 途徑， Fas/FasL 主要介導(dǎo)活

4、化的效應(yīng)性細(xì)胞凋亡，清除激活的外周 CTL，下調(diào)免疫應(yīng) 答。這兩條途徑對(duì)于機(jī)體對(duì)外來抗原進(jìn)行適度的免疫應(yīng)答，維持免疫自穩(wěn)發(fā)揮 重要作用。正常狀態(tài)下，特異性淋巴細(xì)胞在外周免疫器官接受抗原刺激而大量 增殖，在免疫應(yīng)答末期，機(jī)體通過 Fas/FasL 細(xì)胞凋亡機(jī)制使發(fā)生活化的 T 細(xì)胞 凋亡，從而對(duì)免疫過程進(jìn)行嚴(yán)密調(diào)控，以維持免疫自穩(wěn)。因此，TCE 藥疹樣皮 炎的的發(fā)生可能是由于機(jī)體免疫自穩(wěn)狀態(tài)被打破，發(fā)生過強(qiáng)的免疫應(yīng)答反應(yīng)所 致。 為驗(yàn)證上述假說，本研究采用成組設(shè)計(jì)的多個(gè)樣本比較分析以及 TCE 藥疹 樣皮炎患者入院治療以及肝功能恢復(fù)正常時(shí)機(jī)體細(xì)胞免疫學(xué)指標(biāo)的比較分析， 進(jìn)一步探討機(jī)體細(xì)胞免疫學(xué)指

5、標(biāo)的變化與 TCE 藥疹樣皮炎的關(guān)系。研究結(jié)果如 下： 一、一、TCETCE 藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究藥疹樣皮炎細(xì)胞免疫學(xué)指標(biāo)及發(fā)病機(jī)制研究 本研究選擇三組研究對(duì)象，病例組為 16 例 TCE 藥疹樣皮炎患者。接觸對(duì)照 組為 32 例來自與病例同車間的從事 TCE 作業(yè)的健康職業(yè)工人；非接觸對(duì)照組為 28 例從未接觸過 TCE 及其他職業(yè)性有害因素的的健康就業(yè)前體檢工人。三組研 究對(duì)象按年齡性別進(jìn)行匹配后，結(jié)果如下： 1.1.淋巴細(xì)胞抗原特異性增殖功能淋巴細(xì)胞抗原特異性增殖功能 16 例病人除皮疹外，還伴有不同程度的肝損害，谷丙轉(zhuǎn)氨酶（ALT）水平 升高者 15 例（93.33%

6、）；谷草轉(zhuǎn)氨酶（AST）升高者 12 例（73.33%）。三組研 究對(duì)象外周血淋巴細(xì)胞分別與不同濃度 TCE（0.2mmol/L、1 mmol/L、5 mmol/L）共培養(yǎng)，在各組內(nèi)隨著 TCE 染毒濃度的增加外周血淋巴細(xì)胞的存活率 逐漸降低，差異有統(tǒng)計(jì)學(xué)意義(p0.05)，提示 TCE 本身具有細(xì)胞毒性，而在 TCE 藥疹樣 皮炎患者以及 TCE 接觸對(duì)照體內(nèi)并不存在針對(duì) TCE 原形的特異性記憶性淋巴細(xì) 胞。 2.2.外周血淋巴細(xì)胞亞群分析外周血淋巴細(xì)胞亞群分析 研究對(duì)象血常規(guī)檢測(cè)結(jié)果發(fā)現(xiàn)，16 例病人中有 6 例（37.5%）病人外周血 淋巴細(xì)胞（LYMP）高于臨床正常參考值范圍，1（6

7、.25%）例低于臨床正常參考 值范圍；6 例（37.5%）單核細(xì)胞（MONO）高于臨床正常參考值范圍；14 例 （87.5%）中性粒細(xì)胞（NEUT）高于臨床正常參考值范圍；8 例（50%）嗜酸性 粒細(xì)胞（EO）高于臨床正常參考值范圍，2 例（12.5%）低于臨床正常參考值范 圍，8 例（50%）嗜堿性粒細(xì)胞（BASO）高于臨床正常參考值范圍。在接觸對(duì)照 組和非接觸對(duì)照組體內(nèi)上述指標(biāo)均在臨床正常參考值范圍內(nèi)。提示 TCE 藥疹樣 皮炎患者體內(nèi)免疫細(xì)胞的數(shù)量發(fā)生異常，由于外周血免疫細(xì)胞的改變與組織內(nèi) 免疫細(xì)胞的浸潤密切相關(guān)，尤其是嗜酸性粒細(xì)胞主要分布于人體內(nèi)呼吸道、消 化道和泌尿生殖道粘膜組織中，

8、其水平的升高可能與 TCE 藥疹樣皮炎病人出現(xiàn) 的眼、口、生殖器等處的粘膜損傷有關(guān)。 結(jié)合血常規(guī)與流式細(xì)胞術(shù)檢測(cè)的結(jié)果，比較各組之間淋巴細(xì)胞亞群在外周 血中的變化情況。結(jié)果發(fā)現(xiàn)病例組外周血淋巴細(xì)胞總數(shù)、T 細(xì)胞、CD4+T 細(xì)胞計(jì) 數(shù)均明顯高于接觸對(duì)照組和非接觸對(duì)照組(p0.05)，提示這三個(gè)指標(biāo)可作為 TCE 藥疹樣皮炎的疾 病標(biāo)志； CD8+T 細(xì)胞和 CD8+CD28-T 細(xì)胞三組之間比較為病例組接觸對(duì)照組非 接觸對(duì)照組，各組間差異有統(tǒng)計(jì)學(xué)意義(p0.05)，CD3-CD56+細(xì)胞三組之間比較 為病例組接觸對(duì)照組非接觸對(duì)照組，各組之間有統(tǒng)計(jì)學(xué)差異(p0.05)。提示 這三個(gè)指標(biāo)可作為 T

9、CE 接觸工人的免疫學(xué)效應(yīng)指標(biāo)。 3.3.外周血外周血 Fas/FasLFas/FasL、PF/GrBPF/GrB 的測(cè)定的測(cè)定 病例組外周血 CD8+T 細(xì)胞表面 Fas 陽性表達(dá)的百分比高于接觸對(duì)照組和非 接觸對(duì)照組，差異有統(tǒng)計(jì)學(xué)意義（p0.05） ；病例組與接觸對(duì)照組外周血 CD4+T 細(xì)胞表面 FasL 陽性表達(dá)的百分比均高于非接觸對(duì)照組（p0.05） ；CD8+T 細(xì)胞表面 FasL 陽性表達(dá)的百分比呈現(xiàn)病例組接 觸對(duì)照組非接觸對(duì)照組，組間差異有統(tǒng)計(jì)學(xué)意義（p0.05） 。PF 蛋白在外周血 淋巴細(xì)胞中表達(dá)高于接觸對(duì)照與非接觸對(duì)照組(p0.05)。GrB 在病例組的表達(dá)高于非接觸對(duì)照

10、 組 (p0.05)。提示在 病例組工人體內(nèi)活化的 CTL 通過 Fas/FasL 途徑清除了一部分，但由于同時(shí)表達(dá) FasL 的 CD8+CTL 隨血液循環(huán)遷移至皮膚、肝臟、腎臟等組織器官，通過與這些 組織細(xì)胞表面的 FAS 蛋白結(jié)合介導(dǎo)靶細(xì)胞的凋亡。同時(shí)病例組體內(nèi)通過活化的 CTL 高表達(dá) PF 和 GrB 蛋白，介導(dǎo)皮膚角質(zhì)細(xì)胞的死亡，引起 TCE 藥疹樣皮炎患 者皮膚的損傷。而健康的 TCE 接觸對(duì)照工人體內(nèi)也存在活化的細(xì)胞毒性 T 細(xì)胞 （CD4+CTL 與 CD8+CTL） ，但并沒有發(fā)病，其原因仍有待進(jìn)一步研究。 病例組外周血 Fas mRNA 表達(dá)高于接觸對(duì)照組（1.2622

11、倍）與非接觸對(duì)照 組（1.1383 倍）,接觸對(duì)照組低于非接觸對(duì)照組（0.9019 倍）；病例組外周血 FASL mRNA 表達(dá)高于接觸對(duì)照組（1.4239 倍）與非接觸對(duì)照組（1.7507 倍）， 接觸對(duì)照組低于非接觸對(duì)照組（0.8133 倍）。病例組（0.8166 倍）與接觸對(duì)照 組（0.6009 倍）PF mRNA 表達(dá)低于非接觸對(duì)照組，而病例組高于接觸對(duì)照組 （1.3590 倍）。病例組 GrB mRNA 表達(dá)低于接觸對(duì)照組（0.4477 倍）與非接觸 對(duì)照組（0.4835 倍），接觸對(duì)照組高于非接觸對(duì)照組（1.0800 倍）。此研究結(jié) 果與 FAS、FASL、PF、GrB 蛋白的表達(dá)

12、情況不完全一致，是由于 mRNA 的表達(dá)在 轉(zhuǎn)錄和翻譯水平均受到多種因素的調(diào)節(jié)。 二、二、8 8 例例 TCETCE 藥疹樣皮炎病例報(bào)告分析藥疹樣皮炎病例報(bào)告分析 對(duì) 8 例 TCE 藥疹樣皮炎的病人入院治療時(shí)及肝功能恢復(fù)正常時(shí)（轉(zhuǎn)歸時(shí)） 各指標(biāo)進(jìn)行比較分析，進(jìn)一步探索 TCE 藥疹樣皮炎病人體內(nèi)各免疫學(xué)指標(biāo)的變 化以及可用于評(píng)價(jià)臨床治療效果的指標(biāo)，并結(jié)合前面病例與對(duì)照實(shí)驗(yàn)研究結(jié)果 對(duì) TCE 藥疹樣皮炎與各細(xì)胞免疫學(xué)指標(biāo)的關(guān)系進(jìn)行綜合分析。 1.1.各群淋巴細(xì)胞檢測(cè)分析各群淋巴細(xì)胞檢測(cè)分析 獲得 6 例病人入院治療時(shí)及轉(zhuǎn)歸時(shí)淋巴細(xì)胞檢測(cè)結(jié)果資料。與臨床正常參 考值相比較，6 例（100%）病

13、人入院時(shí) NEUT 均升高，1 例病人肝功能恢復(fù)正常 時(shí) NEUT 仍高于正常參考值范圍；入院時(shí) EO 升高者有 4 例（66.67%）；BASO 和 LYMPH 升高者分別有 3 例（50%）；MONO 升高者有 2 例(33.33%)，出院時(shí)仍有 1 例病人 MONO 升高。 對(duì) 6 例病人入院和轉(zhuǎn)歸時(shí)血常規(guī)結(jié)果進(jìn)行比較，其中 6 例（100%）病人入 院時(shí) NEUT 與 BASO 均升高，5 例（83.33%）病人 LYMPH、MONO、EO 均升高。提 示 TCE 藥疹樣皮炎患者在發(fā)病的初期機(jī)體正在進(jìn)行著免疫反應(yīng)，與第一部分研 究結(jié)果相一致。 對(duì) 6 例病人入院和轉(zhuǎn)歸時(shí)淋巴細(xì)胞亞群分析

14、結(jié)果進(jìn)行比較，6 例病人中有 4 例（66.67%）在入院時(shí) T 淋巴細(xì)胞升高，B 淋巴細(xì)胞有 3 例（50%）升高，4 例（66.67%）NK 細(xì)胞降低。入院時(shí) 4 例（66.67%）CD4+T 淋巴細(xì)胞明顯增多，4 例（66.67%）CD8+T 淋巴細(xì)胞明顯增多。CD8+CD28+T 有 3 例（50%）入院時(shí)降低， CD8+CD28-T 有 5 例（83.33%）入院時(shí)升高。6 例病人中有 4 例在入院時(shí) CD4+/CD8+比值降低，4 例 CD28+/CD28-細(xì)胞的比值降低。提示 TCE 藥疹樣皮炎患 者在發(fā)病初期機(jī)體內(nèi)各群免疫細(xì)胞的數(shù)量與比例發(fā)生變化。 2.2.外周血外周血 Fas

15、/FasLFas/FasL、PF/GrBPF/GrB 的測(cè)定的測(cè)定 與疾病轉(zhuǎn)歸時(shí)相比較，入院時(shí) 6 例病人中有 4 例（66.67%）Fas 表達(dá)陽性 CD4+T 細(xì)胞百分比升高；3 例(50%)病人 Fas 陽性 CD8+T 細(xì)胞百分比升高。6 例 病人中有 3 例(50%) FasL 陽性 CD4+T 細(xì)胞升高；3 例(50%)病人 FasL 陽性 CD8+T 細(xì)胞升高。而兩例入院檢測(cè)前未經(jīng)激素治療的病例外周血入院時(shí) Fas 表達(dá) 陽性 CD4+T 細(xì)胞百分比降低、Fas 陽性 CD8+T 細(xì)胞百分比升高、FasL 陽性 CD8+T 細(xì)胞升高，提示病例在發(fā)病的早期 CD4+CTL 未發(fā)揮負(fù)

16、調(diào)節(jié)免疫應(yīng)答的作用， 未及時(shí)清除活化的 T 細(xì)胞。CD8+CTL 通過 Fas/FasL 途徑介導(dǎo)了 TCE 藥疹樣皮炎 患者的病例損傷。3 例(100%)病人在外周血中 PF 表達(dá)陽性淋巴細(xì)胞百分率均高 于出院時(shí)。1 例(33.33%)病人外周血中 GrB 表達(dá)陽性細(xì)胞百分率高于出院時(shí)。 提示在 TCE 藥疹樣皮炎的發(fā)病早期，PF 通過殺傷皮膚角質(zhì)細(xì)胞介導(dǎo) TCE 藥疹樣 皮炎患者的皮膚損害。此與第一部分研究結(jié)果相一致。 總之，本研究通過病例與對(duì)照實(shí)驗(yàn)研究以及病例入院和轉(zhuǎn)歸時(shí)細(xì)胞免疫學(xué) 指標(biāo)的比較分析，揭示 TCE 藥疹樣皮炎的發(fā)生可能是由其代謝產(chǎn)物而非 TCE 本 身引起，TCE 藥疹樣皮

17、炎的發(fā)生與體內(nèi)免疫細(xì)胞的活化增殖有關(guān)，TCE 暴露能誘 導(dǎo)人外周血淋巴細(xì)胞亞群的數(shù)量發(fā)生變化，其中總淋巴細(xì)胞、T 細(xì)胞和 CD4+T 細(xì) 胞計(jì)數(shù)升高可能作為 TCE 接觸工人發(fā)生藥疹樣皮炎的的臨床篩檢指標(biāo)，CD3- CD56+細(xì)胞計(jì)數(shù)降低、CD8+T 細(xì)胞計(jì)數(shù)升高以及 CD8+CD28-T 計(jì)數(shù)升高可作為反映 TCE 接觸的效應(yīng)指標(biāo)。CD4+CTL 細(xì)胞通過表達(dá) Fas 介導(dǎo)活化的 T 細(xì)胞凋亡，維持 機(jī)體的免疫自穩(wěn)。CD8+CTL 通過表達(dá) FasL 并隨血液循環(huán)與皮膚、肝臟、腎臟等 組織細(xì)胞表面 Fas 結(jié)合介導(dǎo)靶細(xì)胞凋亡，引起組織器官的免疫性損傷。CTL 通 過 PF/GrB 途徑介導(dǎo)

18、TCE 藥疹樣皮炎患者的皮膚損害。因此本研究為 TCE 接觸工 人的健康監(jiān)護(hù)具有重要指導(dǎo)意義和參考價(jià)值，并對(duì) TCE 藥疹樣皮炎病人采取針 對(duì)性的免疫治療措施提供理論依據(jù)。 關(guān)鍵詞：關(guān)鍵詞：三氯乙烯，細(xì)胞免疫，藥疹，接觸性過敏性皮炎，半抗原，細(xì)胞毒性 T 細(xì)胞，淋巴細(xì)胞亞群，職業(yè)接觸 Study on Index of Cell-mediated immunity and Pathogenesis of Dermatitis medicamentosa-like of trichloroethylene 英文摘要英文摘要 Abstract Dermatitis medicamentosa-li

19、ke of trichloroethylene(DMLT) in workers appears to be an occupational health issue and has aroused general concern in China in recent years. However, little is known about its pathogenesis to date, there are also lack of specific indexes for health surveillance as well as specific measures for clin

20、ical treatment. DMLT may involve in type hypersensitivity according to its characteristics. Formation of T memory lymphocytes and proliferation are the two steps of cell-mediated immunity. Activated T cells regulate the proliferation of NK and B cells through secrete a series of cytokines. Cytotoxic

21、 T lymphocytes(CTL) play an important part in cell-mediated immune system. Its cytotoxicity can be induced by two ways: a) granuledependent exocytosis pathway, which participate immune defenses. b) Fas-FasL intercellular linkage-mediated pathway, which down-regulate immune response through clean act

22、ivated peripheral CTL at the end of the immune response. The two ways play an important role in maintaining the immune homeostasis. The process of DMLT may related to the proliferation of lymphocytes and the function of CTL. Those two approaches play an important role in cleaning foreign antigens to

23、 maintain immune homeostasis. In normal condition, specific immune lymphocytes which stimulated by antigen proliferate in peripheral immune organs. In the end of the immune response, the activated cells apoptosis through Fas/FasL pathway, thus to control the immune process and maintain immune homeos

24、tasis. Therefore, DMLT might be induced by activated cells which were not cleared in time, and strong immune response breakout. In order to test the above hypothesis, we conducted a three groups design analysis and cases analysis to explore the relationship between abnormal index of cell-mediated im

25、munity and DMLT. The results were as follows: 1. Study on Index of Cell-mediated immunity and Pathogenesis of DMLT There are there groups in this study: case group, cases are 16 patients of DMLT, all examinations executed on admission; TCE- exposure group, 30 workers came form the workshops the case

26、s occurred but no skin disorders examined by the occupational physicians after the first 3 months of TCE exposure. Healthy worker group, 28 workers were never exposed to TCE and no history of previous skin disease. The main results are as follows: .Test for functions of antigen-specific lymphocyte p

27、roliferation 16 patients with severe rash, accompanied by different degree of liver damage, 15 cases (93.33%) in 16 with higher ALT and 12 cases (73.33%)in 16 with higher AST. Peripheral blood lymphocytes(2106/ml) of three groups cultured with different concentrations TCE (0.2mmol/L,1mmol/L, 5mmol/L

28、). In any group, peripheral blood lymphocyte survival rates gradually reduced with the increase of TCE concentration. No significant difference was found about survival rates of lymphocytes under the same concentrations in three groups. The result suggest that TCE performed great cell toxicity on pe

29、ripheral blood lymphocytes, and there was no TCE-specific memory lymphocyte. . Analysis of lymphocyte subsets in peripheral blood From the routine blood test of 16 patients, 6 cases(37.5%) peripheral blood lymphocyte(LYMP) higher than the normal clinical range, 1(6.25%) below the normal clinical ran

30、ge; 6 cases(37.5%) mononuclear cells (MONO) higher than normal reference range of clinical; 14 cases(87.6%) neutrophils NEUT) higher than normal clinical range; 8 cases(50%) eosinophils(EO) higher than normal clinical range, 2 cases (12.5%) below the normal clinical range; 8 cases (50%) basophilic g

31、ranulocyte (BASO) higher than the normal range of clinical range. In both healthy TCE-exposed workers and unexposed workers, the indexes are all in clinical normal reference ranges. Those indicate immune response on going in patients of DMLT. The change of peripheral immune cells are closely related

32、 to the organization of immune cell infiltrates, human eosinophils mainly distributed in the respiratory tract, enteron and genitourinary tract mucosa tissue, the increase of EO in peripheral blood might relate to the damage of mucous membrane in eye, mouth and genital of patients. The absolute coun

33、ts of lymphocyte, T cell, CD4+ T cell were significantly increased in patients with hypersensitivity dermatitis compared with healthy TCE-exposed workers and unexposed workers(P0.05), so those three indexes could be disease markers of DMLT. CD8+ T cell, CD8+CD28- T cell counts among the three groups

34、 showed case groupexposed groupunexposed control group, and the difference was significant(P0.05). NK cell counts among the 3 groups showed the case groupexposed groupunexposed control group, and the difference was significant(P0.05), so those three indexes could be immunology effect makers for TCE

35、exposure. .Examination of Fas/FasL、PF/GrB expression of protein and mRNA in peripheral blood The percentage of CD8+CD95+T cell in CD8+T cell in case is higher than exposed group and unexposed group(P0.05). The percentage of CD4+CD178+T cell in CD4+T cell in case and exposed group are higher than une

36、xposed group(P0.05), no significant difference between case group and unexposed group(P0.05).The percentage of CD8+CD178+T cell in CD8+T cell among the three groups showed case groupexposed groupunexposed control group, and the difference was significant (P0.05). In case group, PF protein in periphe

37、ral lymphocyte is higher than exposed group and unexposed group(P0.05). It indicate CTL (CD4+CTL and CD8+CTL) induce apoptosis of effector cells in healthy TCE-exposed workers. However, in cases, some CTLs are removed through Fas/FasL pathway, others migrate to skin, liver, kidney, etc, and mediated

38、 the target cell apoptosis. Meanwhile, the high expression of PF and GrB cause skin damage in cases. In case group the expression of FAS mRNA is higher than exposed(1.2622 fold) and unexposed group(1.1383 fold), exposed group is lower than unexposed group(0.9019 fold); In case group the expression o

39、f FasL mRNA is higher than exposed(1.4239 fold) and unexposed group(1.7507 fold), exposed group is lower than unexposed group(0.8133 fold); In case (0.8133 fold)and exposed group (0.6009 fold)the expression of PF mRNA are lower than unexposed group, in case group (1.3590 fold)is higher than unexpose

40、d group. In case group the expression of GrB mRNA is lower than exposed(0.4477 fold) and unexposed group(0.4835 fold), exposed group(1.0800 fold) is higher than unexposed group. The expression of FAS, FASL, PF, GrB in protein and mRNA level are not identical, that is due to the expression of mRNA in

41、 transcription and translation are regulated by various factors. 2. 8 cases report of DMLT Compare the differences of DMLT when hospitalized and liver function recovered to normal, further explore the indexes of immunological changes and that could be used to evaluate the clinical treatment effect,

42、and combining with the former group analysis to analyze the relationship between immunological indexes and DMLT. .Analysis of lymphocyte subsets There are lymphocyte results of six patients when hospitalized and liver function recovered to normal. NEUT in 6 cases(100%) were higher when checked into

43、hospital; 4 in 6 cases(66.67%)had a higher EO than normal clinical range; 3 in 6(50%) cases had a higher BASO and LYMPH than normal clinical range. 6 cases(100%) had higher NEUT and BASO, 5 cases in 6 had higher LYMPH、MONO and EO when hospitalized than liver function recovered to normal. Those indic

44、ate immune function disorder in DMTL, all consistent with the former results. Lymphocyte subsets analysis results of six patients in hospital and liver back to normal, 4 cases (66.67%)in 6 have significantly higher T lymphocytes in admission; B lymphocytes, 3 cases (50%) was obviously higher than no

45、rmal liver, 4 cases (66.67%) NK cell below normal liver. On admission, 4 cases (66.67%) CD4+T lymphocyte increased obviously, 4 cases (66.67%) CD8+T lymphocytes increased obviously. In 3(50%)cases CD8+CD28+T below in admission, 5 cases (83.33%) CD8+CD28-T higher than liver back to normal. 4 patients

46、 in 6 patients (66.67%) CD4+/CD8+ ratio decreased when hospitalized, 4 cases CD28+/CD28- cell ratio decreased. It indicated there are changes of immune cells number and proportion onset of DMLT. Case 1, 5, and 6 discharged from hospital when the liver function returning to normal. NK cells elevated、

47、 CD28+CD8-T cells reduced and the ratio of CD4+/CD8+ returned to normal in 3 patients when discharged from hospital. Those three indexes could be the index of rehabilitation. In case 1 and case 6, T lymphocytes、CD8+T lymphocytes declined when discharged from hospital, it might be a reference index o

48、f rehabilitation. .Examination of Fas/FasL、PF/GrB expression of protein and mRNA in peripheral blood 4 patients in 6 (50%) the percentage of Fas+CD4+T cells in CD4+T were higher when hospitalized 、3 patients (50%) patients the percentage of FAS+CD8+Tcells in CD8+T were higher at admission than norma

49、l liver function, 3 patients in 6 (50%) the percentage of FasL+CD4+T cells in CD4+T were lower, 3 cases (50%) the percentage of FasL+CD8+T cells in CD8+T were lower than rehabilitation. Those results are consistent with the former results. PF protein in peripheral lymphocyte was higher in 3 cases(10

50、0%) when hospitalized than liver function recovered to normal; 1 in 3 cases(33.33%) GrB protein in peripheral lymphocyte was higher when hospitalized than liver function recovered to normal. It indicated PF damage skin keratinized cell at the early stage of DMLT. In summary, this study revealed the

51、immune response in DMLT might induced by the metabolites of TCE rather than TCE itself. There were great changes in DMLT about lymphocyte subsets in peripheral blood. Lymphocyte, T cell, CD4+T cell could be disease markers of DMLT, CD8+T, CD8+CD28- T and NK cell could be effect makers for TCE exposu

52、re. CTLs take part in the damage of skin 、liver、kidney and some other organs. Those results could make some significance to health surveillance of TCE exposure and provide possibility for further studies in pathogenesis. KeyKey words:words: trichloroethylene，cell-mediated immunity，drug rash，allergic

53、 contact dermatitis，hapten，cytotoxic T lymphocyte，Lymphocyte subsets, occupational exposure 英文縮寫詞表英文縮寫詞表 （ListList ofof AbbreviationAbbreviation） ACDACDallergicallergic aminotransferaseaminotransferase 過敏性接觸性皮炎過敏性接觸性皮炎 ADADAtopicAtopic dermatitisdermatitis 特應(yīng)性皮炎特應(yīng)性皮炎 ADHADHalcoholalcohol dehydrogena

54、sedehydrogenase 醇脫氫酶醇脫氫酶 ALTALTglutamic-pyruvicglutamic-pyruvic transaminasetransaminase 谷丙轉(zhuǎn)氨酶谷丙轉(zhuǎn)氨酶 ALDHALDHaldehydedehydrogenasealdehydedehydrogenase 醛脫氫酶醛脫氫酶 ASTASTglutamic-oxaloaceticglutamic-oxaloacetic transaminasetransaminase 谷草轉(zhuǎn)氨酶谷草轉(zhuǎn)氨酶 BASOBASObasophilsbasophils 嗜堿性粒細(xì)胞嗜堿性粒細(xì)胞 CDCDclustercluste

55、r ofof differentiationdifferentiation 分化抗原分化抗原 CHSCHSContactContact hypersensitivityhypersensitivity 接觸性超敏反應(yīng)接觸性超敏反應(yīng) CTLCTLcytotoxiccytotoxic T T lymphocytelymphocyte 細(xì)胞毒性細(xì)胞毒性 T T 細(xì)胞細(xì)胞 CYP450CYP450CytochtomeCytochtome P450P450 細(xì)胞色素細(xì)胞色素 P450P450 酶系酶系 DCVGDCVG(1,2-dichlorovinyl)glutathione(1,2-dichloro

56、vinyl)glutathione 1,2-1,2-二氯乙烯基谷胱甘肽二氯乙烯基谷胱甘肽 DCVCDCVC(1,2-dichlorovinyl)-L-cysteine(1,2-dichlorovinyl)-L-cysteine 1,2-1,2-二氯乙烯基半胱氨酸二氯乙烯基半胱氨酸 DMLTDMLT DermatitisDermatitis medicamentosa-medicamentosa- likelike ofof trichloroethylenetrichloroethylene 三氯乙烯藥疹樣皮炎三氯乙烯藥疹樣皮炎 DMSODMSOdimethylsulfoxidedimethy

57、lsulfoxide 二甲基亞砜二甲基亞砜 EDTAEDTA DisodiumDisodium thylenediaminetetrathylenediaminetetra acetateacetate 乙二胺四乙酸乙二胺四乙酸 EOEOeosnophilseosnophils 嗜酸性粒細(xì)胞嗜酸性粒細(xì)胞 FASLFASLfasfas ligandligand 外周血淋巴細(xì)胞外周血淋巴細(xì)胞 FCSFCSfetalfetal calfcalf serumserum 胎牛血清胎牛血清 GGTGGTglutamineglutamine transferasetransferase 谷氨酰胺轉(zhuǎn)移酶谷氨酰

58、胺轉(zhuǎn)移酶 GrBGrBgranzymegranzyme B B 顆粒酶顆粒酶 B B GSHGSHglutathioneglutathione 谷胱甘肽谷胱甘肽 GSTsGSTsGlutathioneGlutathione S-tranferaseS-tranferase 谷胱甘肽轉(zhuǎn)移酶谷胱甘肽轉(zhuǎn)移酶 IARCIARC InternationalInternational agencyagency forfor researchresearch onon cancercancer 國際癌癥研究署國際癌癥研究署 ILILinterleukininterleukin 白細(xì)胞介素白細(xì)胞介素 IFNI

59、FNinterferoninterferon 干擾素干擾素 LYMPLYMPlymphocytelymphocyte countcount 淋巴細(xì)胞計(jì)數(shù)淋巴細(xì)胞計(jì)數(shù) MFIMFImeanmean fluorescentfluorescent intensityintensity 平均熒光強(qiáng)度平均熒光強(qiáng)度 MONOMONOmonocytesmonocytes 單核細(xì)胞單核細(xì)胞 MTTMTTmethylmethyl thiazolylthiazolyl tetrazoliumtetrazolium 噻唑蘭噻唑蘭 NATsNATsN-acetyltranferaseN-acetyltranferas

60、e N-N-乙酰轉(zhuǎn)移酶乙酰轉(zhuǎn)移酶 NEUTNEUTneutrophilsneutrophils 中性粒細(xì)胞中性粒細(xì)胞 NIRSNIRSNonimmediateNonimmediate AllergicAllergic reactionsreactions 非速發(fā)型超敏反應(yīng)非速發(fā)型超敏反應(yīng) NKNK cellcellnaturalnatural killerkiller cellcell 自然殺傷細(xì)胞自然殺傷細(xì)胞 PBSPBSphosphatephosphate bufferedbuffered salinesaline 磷酸鹽緩沖液磷酸鹽緩沖液 PFPFperforinperforin 穿孔素