本文選題：抗CD20單克隆抗體 + B淋巴細(xì)胞��； 參考：《吉林大學(xué)》2012年碩士論文

【摘要】：目的：本實(shí)驗(yàn)探討抗CD20單克隆抗體在小鼠皮膚移植模型中如何預(yù)防急性體液性排異反應(yīng)。目前研究證實(shí)，T淋巴細(xì)胞在急性體液性排異反應(yīng)中起主導(dǎo)作用，而B淋巴細(xì)胞及其成熟漿細(xì)胞產(chǎn)生的抗體參與了體液性排異反應(yīng)的發(fā)生。由于免疫抑制劑高效性和臨床應(yīng)用的廣泛性，使同種異體腎移植進(jìn)步顯著，移植物總體存活時(shí)間明顯延長(zhǎng)，排斥總體發(fā)生率明顯降低。然而，目前臨床上常規(guī)使用的抗增殖類藥物、鈣神經(jīng)蛋白抑制劑及激素主要針對(duì)于T淋巴細(xì)胞介導(dǎo)的細(xì)胞免疫，對(duì)體液性免疫抑制作用遠(yuǎn)低于對(duì)細(xì)胞免疫抑制作用[1]。目前應(yīng)用于臨床的針對(duì)B淋巴細(xì)胞的藥物較少，如何清除B淋巴細(xì)胞，抑制B淋巴細(xì)胞在體液性排異反應(yīng)中發(fā)揮的作用，成為國(guó)內(nèi)外移植中心研究的熱點(diǎn)。由B淋巴細(xì)胞產(chǎn)生的作為參與體液性排異反應(yīng)的供體特異性抗體（donor-specific alloantibodies，DSA）將供者的上皮細(xì)胞作為攻擊的靶點(diǎn)[2]，產(chǎn)生抗體介導(dǎo)的體液性排異反應(yīng)。因此清除受者體內(nèi)的B淋巴細(xì)胞、抗體及DSA成為預(yù)防及治療抗體介導(dǎo)的體液性排異反應(yīng)的焦點(diǎn)。隨著對(duì)B淋巴細(xì)胞及其作用機(jī)制的深入研究證實(shí)，CD20抗原分子是表達(dá)于前B淋巴細(xì)胞及成熟B淋巴細(xì)胞表面的跨膜糖蛋白，是參與B淋巴細(xì)胞活化以及抗原識(shí)別的主要分子�？笴D20單克隆抗體與B淋巴細(xì)胞表面的CD20分子有較高的親和力，可通過誘導(dǎo)細(xì)胞凋亡作用、抗體依賴的細(xì)胞介導(dǎo)的細(xì)胞毒作用及補(bǔ)體介導(dǎo)的細(xì)胞毒作用殺傷B淋巴細(xì)胞。因此，CD20單克隆抗體是一類具有獨(dú)特生物學(xué)作用的生物制劑，在移植抗體介導(dǎo)的體液性排異治療中扮演著重要的角色。 方法：一組受鼠給予供體脾臟細(xì)胞懸液腹腔注射，后提取血清經(jīng)鼠尾靜脈注射法注射到另一只受鼠體內(nèi)；另一組給予抗CD20單克隆抗體鼠尾靜脈注射。然后建立皮膚移植及血管組織皮下包埋模型，術(shù)后觀察移植皮膚存活狀況。應(yīng)用流式細(xì)胞技術(shù)檢測(cè)鼠尾靜脈注射抗CD20單克隆抗體小鼠B淋巴細(xì)胞，并切取移植皮膚及血管組織行組織病理學(xué)檢查，進(jìn)一步研究抗CD20單克隆抗體在抗體介導(dǎo)的體液性排異反應(yīng)中的作用。 結(jié)果： （1）對(duì)照組小鼠移植皮片平均存活時(shí)間（MST）為11.5天，鼠尾靜脈注射血清組的皮片平均存活時(shí)間為9天，鼠尾靜脈注射抗CD20單克隆抗體組的皮片平均存活時(shí)間為12.5天。 （2）流式細(xì)胞技術(shù)檢測(cè)鼠尾靜脈注射抗CD20單克隆抗體組脾細(xì)胞中B淋巴細(xì)胞（B220+）的百分比。與正常小鼠比較，，鼠尾靜脈注射抗CD20單克隆抗體組B淋巴細(xì)胞顯著減少，P0.05。 （3）取3組脾臟組織行HE染色病理學(xué)檢查。鼠尾靜脈注射血清組較對(duì)照組脾臟組織中生發(fā)中心明顯增大，淋巴細(xì)胞密集；鼠尾靜脈注射抗CD20單克隆抗體組較對(duì)照組脾小體明顯縮小，淋巴細(xì)胞明顯減少。 （4）取受體鼠移植皮片及血管組織行HE染色病理學(xué)檢查。鼠尾靜脈注射血清組較對(duì)照組移植皮片組織和皮下包埋血管組織炎細(xì)胞及淋巴細(xì)胞浸潤(rùn)明顯增多，組織結(jié)構(gòu)破壞明顯。血管組織炎癥反應(yīng)重，部分血管壁機(jī)化，血管各層組織破壞嚴(yán)重，管腔堵塞。鼠尾靜脈注射抗CD20單克隆抗體組移植皮片組織和皮下包埋血管組織炎細(xì)胞及淋巴細(xì)胞浸潤(rùn)明顯少于對(duì)照組，組織結(jié)構(gòu)破壞不明顯。血管組織炎癥反應(yīng)輕，血管各層組較完整，管腔存在。 結(jié)論：通過鼠尾靜脈注射抗CD20單克隆抗體組小鼠移植皮片生存時(shí)間延長(zhǎng)，術(shù)前使用抗CD20單克隆抗體能有效預(yù)防抗體介導(dǎo)的急性排異反應(yīng)的發(fā)生。
[Abstract]:Objective : To investigate the effect of anti - CD20 monoclonal antibody on the prevention of acute humoral rejection in mouse skin transplantation model . B - lymphocytes , antibodies and DSA are the main targets for the prevention and treatment of antibody - mediated humoral rejection . As a result , the CD20 monoclonal antibody is a kind of biological agent with unique biological effect , which plays an important role in the humoral rejection therapy mediated by transplantation antibody .

Methods : A group of mice were injected intraperitoneally with the donor spleen cell suspension , and the serum was injected into the other mice via the tail vein injection method .
The effect of anti - CD20 monoclonal antibody on antibody - mediated humoral rejection was investigated .

Results :

( 1 ) The average survival time ( MST ) was 11.5 days in the control group , and the average survival time was 9 days after the tail vein injection . The average survival time of the anti - CD20 monoclonal antibody group was 12.5 days .

( 2 ) The percentage of B lymphocytes ( B220 + ) was detected by flow cytometry in the spleen cells of mouse tail vein anti - CD20 monoclonal antibody group . Compared with normal mice , the anti - CD20 monoclonal antibody group B lymphocytes were significantly decreased in mice tail vein , P0.05 .

( 3 ) HE staining was performed in three groups of spleen tissues .
Compared with control group , the anti - CD20 monoclonal antibody group was significantly reduced in mice tail vein , and the lymphocytes decreased significantly .

( 4 ) HE staining was performed on the skin graft and vascular tissue of the recipient mice . The tissue structure was damaged . The inflammatory response , partial vessel wall movement , tissue destruction in the vascular tissues were significantly smaller than those in the control group . The tissue structure was not clear . The inflammatory reaction of the vascular tissue was mild , the vessel layers were intact , and the lumen was present .

Conclusion : Anti - CD20 monoclonal antibody can be used to prevent antibody - mediated acute rejection by intravenous injection of anti - CD20 monoclonal antibody .
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R392.4

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