本文選題：弓形蟲 + ESA��； 參考：《山西醫(yī)科大學(xué)》2006年碩士論文

【摘要】： 目的本研究首先觀察弓形蟲感染小鼠腹水中分離的排泄-分泌抗原(excreted/secreted antigens ESA)鼻內(nèi)免疫小鼠后對(duì)不同黏膜部位和系統(tǒng)的免疫效應(yīng),又觀察了Vero細(xì)胞株體外培養(yǎng)弓形蟲并提取ESA鼻內(nèi)免疫小鼠后對(duì)不同黏膜部位和系統(tǒng)的免疫效應(yīng)以及誘導(dǎo)免疫應(yīng)答持續(xù)時(shí)間的觀察。探討不同途徑獲得的弓形蟲ESA鼻內(nèi)免疫誘導(dǎo)的免疫應(yīng)答機(jī)制及免疫效果,為弓形蟲鼻內(nèi)復(fù)合疫苗研制奠定理論基礎(chǔ)。 方法本研究分三部分:第一部分觀察弓形蟲感染小鼠腹水中分離的排泄-分泌抗原(ESA)鼻內(nèi)免疫小鼠后對(duì)不同黏膜部位和系統(tǒng)的免疫效應(yīng)。實(shí)驗(yàn)組分別用從感染弓形蟲小鼠后24hESA組、48h ESA組、72h ESA組提取的ESA 20μg/只鼻內(nèi)免疫小鼠2次,間隔2周;對(duì)照組用20μl/只PBS滴鼻。觀察小鼠健康和死亡情況,記錄體重。在末次免疫后第3w處死小鼠,稱取脾臟重量;計(jì)數(shù)派伊爾氏(Peyer’s patches, PP)個(gè)數(shù);分離PP淋巴細(xì)胞、脾淋巴細(xì)胞、小腸上皮內(nèi)淋巴細(xì)胞(intraepithelial lymphocyte, IEL)及腸系膜淋巴結(jié)(mesenteric lymph node, MLN)細(xì)胞并計(jì)數(shù)。眼眶采血和收集直腸內(nèi)糞便,ELISA檢測(cè)糞內(nèi)sIgA水平及血清內(nèi)特異性IgG。 第二部分觀察Vero細(xì)胞株體外培養(yǎng)弓形蟲并提取ESA鼻內(nèi)免疫小鼠后對(duì)不同黏膜部位和系統(tǒng)的免疫效應(yīng)。實(shí)驗(yàn)組分別用5d、7d、9d、11d、14d提取的ESA 20μg/只鼻內(nèi)免疫小鼠2次,間隔2周;對(duì)照組用20μl/只無(wú)攻蟲的細(xì)胞培養(yǎng)上清液滴鼻。觀察小鼠健康和死亡情況,記錄體重。在末次免疫后第3w處死小鼠,稱取脾臟重量;計(jì)數(shù)PP個(gè)數(shù);分離PP、脾淋巴細(xì)胞、腸上皮內(nèi)淋巴細(xì)胞及腸系膜淋巴結(jié)細(xì)胞并計(jì)數(shù)。在二次免疫后第3周處死小鼠,收集糞便和眼眶采血,ELISA檢測(cè)糞內(nèi)sIgA水平及血清內(nèi)特異性IgG。 第三部分本研究觀察了Vero細(xì)胞株體外培養(yǎng)弓形蟲并提取的ESA鼻內(nèi)免疫BALB/c小鼠后不同黏膜部位和系統(tǒng)的免疫效應(yīng)及其持續(xù)時(shí)間。實(shí)驗(yàn)組以免疫原性好的ESA(20μg/只)為抗原,鼻內(nèi)免疫,對(duì)照組用未接種弓形蟲的細(xì)胞培養(yǎng)上清液20μl/只滴鼻。滴鼻2次(間隔2周)后分別于第1、2、3、4、5、6、7周處死小鼠。ELISA法測(cè)定血清IgG、IgA;糞便sIgA及腸液sIgA;稱取脾臟重量;計(jì)數(shù)PP個(gè)數(shù);分離PP、脾、腸上皮內(nèi)淋巴細(xì)胞(intestinal intraepithelial lymphocytes, IEL)并計(jì)數(shù)。 結(jié)果弓形蟲感染小鼠腹水中分離的排泄-分泌抗原(ESA)鼻內(nèi)免疫小鼠后72h ESA組小鼠健康狀況差,體重逐漸降低,死亡4只;其它組小鼠體重仍呈增高趨勢(shì)。實(shí)驗(yàn)各組小鼠IEL數(shù)、脾淋巴細(xì)胞數(shù)、MLN淋巴細(xì)胞數(shù)均高于對(duì)照組(P0.05),有較高的增殖活性。其中72h ESA組和48h ESA組的IEL數(shù)、脾淋巴細(xì)胞數(shù)、MLN細(xì)胞數(shù)顯著高于24h ESA組(P0.01)。弓形蟲ESA免疫后第3周實(shí)驗(yàn)各組小鼠的血清IgG水平、糞便sIgA水平均高于對(duì)照組(P0.05),其中72h ESA組和48h ESA組的抗體水平顯著高于第24h ESA組(P0.01)。
[Abstract]:Objective to investigate the immunological effects of excreted-secreted antigens isolated from ascites of mice infected with Toxoplasma gondii (Toxoplasma gondii) on different mucosal sites and systems after nasal immunization with Toxoplasma gondii (Toxoplasma gondii) infected mice. The immune effects of Toxoplasma gondii (Toxoplasma gondii) on different mucosal sites and systems and the duration of induction of immune response were observed in mice immunized with ESA after nasal immunization with Toxoplasma gondii in vitro. To study the immune response mechanism and immune effect induced by different ways of intranasal immunization of Toxoplasma gondii ESA, and to lay a theoretical foundation for the development of Toxoplasma gondii intranasal compound vaccine. Methods the present study was divided into three parts: the first part was to observe the immune effects on different mucosal sites and systems of mice immunized with excretion-secretory antigen (ESA) isolated from ascites infected by Toxoplasma gondii (Toxoplasma gondii). Mice in the experimental group were immunized with ESA 20 渭 g / mouse after infection with Toxoplasma gondii (Toxoplasma gondii) for 48 h, ESA group (72 h) and control group (20 渭 l / PBS). The health and death of mice were observed and their body weight was recorded. At the third week after the last immunization, the mice were killed, the weight of spleen was taken, the number of Peyers patcheses (PPs) was counted, PP lymphocytes, spleen lymphocytes, intraepithelial lymphocytes (IELs) and mesenteric lymph node, MLN) cells of mesenteric lymph nodes were isolated and counted. SIgA levels in feces and serum specific IgGs were detected by Elisa in orbital blood collection and rectum stool collection. The second part was to observe the immune effects of Toxoplasma gondii (Toxoplasma gondii) cultured in vitro on different mucosal sites and systems in mice immunized with ESA. The mice in the experimental group were immunized with ESA 20 渭 g / mouse for 2 weeks, respectively, and the control group was infused with 20 渭 l / a cell culture supernatant. The health and death of mice were observed and their body weight was recorded. The mice were killed at the third week after the last immunization, the weight of spleen was taken, the number of PP was counted, and PPP-spleen lymphocytes, intestinal intraepithelial lymphocytes and mesenteric lymph node cells were isolated and counted. The mice were killed at the third week after the second immunization. The fecal and orbital blood samples were collected to detect the level of sIgA in feces and the specific IgGs in serum by enzyme-linked immunosorbent assay (Elisa). In the third part, the immune effects and duration of different mucosal sites and systems of Vero cell line cultured in vitro and extracted ESA from BALB/c mice were observed. The experimental group was immunized with ESA(20 渭 g / mouse, and the control group was inoculated with 20 渭 l of the culture supernatant of Toxoplasma gondii cells. The mice were killed at 7 weeks after nasal drip twice (interval 2 weeks). Elisa method was used to determine the serum IgGG IgA, the fecal sIgA and intestinal Siga, the weight of spleen, the number of PP, the separation of PPp, spleen and intestinal intraepithelial lymphocytes, and count. Results the mice inoculated with Toxoplasma gondii in ascites with excretion-secretory antigen (ESAA) were in poor health condition, their body weight gradually decreased, and 4 mice died, while the other groups showed an increasing trend of body weight. The number of IEL and spleen lymphocytes in each group was higher than that in control group (P 0.05), and it had higher proliferative activity. The number of IEL and spleen lymphocytes in 72 h ESA group and 48 h ESA group were significantly higher than that in 24 h ESA group. The levels of serum IgG and fecal sIgA were significantly higher in Toxoplasma gondii ESA group than those in control group at 3 weeks after ESA immunization. The antibody levels in 72 h ESA group and 48 h ESA group were significantly higher than those in 24 h ESA group.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 吳靜;ESA和STAg鼻內(nèi)免疫小鼠誘導(dǎo)的抗弓形蟲感染保護(hù)性免疫應(yīng)答[D];山西醫(yī)科大學(xué);2007年

，

本文編號(hào)：1897624