本文選題：剛地弓形蟲 + 弓形蟲病�。� 參考：《山西醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 目的本研究首先建立經(jīng)口感染弓形蟲RH株速殖子垂直傳播小鼠模型,然后觀察可溶性速殖子抗原(soluble tachyzoite antigen,STAg)滴鼻免疫小鼠對弓形蟲垂直傳播的阻斷作用,探討STAg聯(lián)合排泄.分泌抗原(excreted/secreted antigens,ESA)滴鼻免疫小鼠對弓形蟲垂直傳播的阻斷作用,為研制阻斷弓形蟲垂直傳播疫苗提供實驗依據(jù)。 方法本研究分三部分:第一部分:54只妊娠0天的BALB/c孕鼠隨機分為6組,于妊娠第8天,分別用0、1000、2000、4000、8000、16000個弓形蟲速殖子灌胃感染,妊娠第18天處死,記錄活胎率,測定母鼠肝、胎盤和胎仔腦組織弓形蟲抗原。 第二部分:130只5～6周齡BALB/c處女鼠隨機分為2個大組:PBS組和STAg組。PBS組以PBS 20μl/只滴鼻,STAg組以STAg 20μg/只滴鼻,免疫2次,間隔2周。末次免疫后處女鼠與雄鼠以2∶1分批同籠交配,各組按免疫后攻蟲時間不同再分為1,2,3組,每組10只。妊娠第8天(分別對應(yīng)末次免疫后第11天、第17天、第23天)用8000個/只速殖子灌胃攻擊小鼠。妊娠第18天頸椎脫臼處死,記錄孕鼠感染率、活胎率,稱重脾臟、胸腺重量,采集母鼠腸液進行IgA檢測,測定母鼠肝、胎盤和胚胎腦組織弓形蟲抗原。 第三部分:BALB/c處女鼠40只隨機分為4組:PBS組、STAg組、ESA組和聯(lián)合抗原(STAg+ESA)組,每組10只。STAg組、ESA組和聯(lián)合抗原(STAg+ESA)組分別用20μg/只STAg、20μg/只ESA和20μg/只聯(lián)合抗原(10μg STAg+10μg ESA)滴鼻,PBS組用PBS以20μl/只滴鼻,免疫2次,間隔2周。末次免疫后第13天處女鼠與雄鼠以2∶1分批進行同籠交配。妊娠第8天用8000個速殖子/只灌胃攻擊全部小鼠。妊娠第18天頸椎脫臼處死,記錄孕鼠感染率、活胎率,計數(shù)脾組織內(nèi)弓形蟲速殖子,檢測母鼠小腸沖洗液sIgA和血清IgG抗體水平,測定母鼠肝、胎盤和胎仔腦組織弓形蟲抗原。 結(jié)果不同劑量弓形蟲經(jīng)口感染孕鼠后,孕鼠死亡率與胎盤和胚胎感染率均隨感染劑量的增加而升高,而活胎率下降。1000、2000組的胎盤、胚胎未發(fā)生感染,4000組的胚胎感染率5.13%,16000組的胚胎感染率20.00%,但孕鼠的健康狀況差、死亡率高,均不適宜建立弓形蟲垂直傳播動物模型。而8000組孕鼠既有較高的存活率,又可保證胎盤和胚胎較高的感染率。 用可溶性速殖子抗原(soluble tachyzoite antigen,STAg)鼻內(nèi)免疫小鼠后,STAg第1、2、3組和PBS第1、2、3組孕鼠感染率和死亡率無統(tǒng)計學(xué)差異。STAg第3組活胎率要明顯高于PBS第1組、2組和3組(P＜0.05)。各組脾臟重量未見明顯變化;STAg第3組小鼠胸腺重量低于其它組(P＜0.05)。STAg第3組孕鼠腸液sIgA高于除STAg第2組外的其它組(P＜0.05);STAg第1組、2組、3組孕鼠感染率隨免疫時間的增加而減少,而活胎率增高,胎盤、胚胎感染率隨免疫時間的增加而減少。STAg第3組的胚胎感染率低于其它組。 可溶性速殖子抗原(soluble tachyzoite antigen,STAg)聯(lián)合排泄-分泌抗原(excreted/secreted antigens,ESA)鼻內(nèi)免疫小鼠后,STAg組、ESA組和聯(lián)合抗原組孕鼠感染率低于PBS組,但無統(tǒng)計學(xué)差異。PBS組活胎率顯著低于ESA組和聯(lián)合抗原組(P＜0.01)。STAg組、ESA組和聯(lián)合抗原組脾組織內(nèi)蟲荷(速殖子數(shù))顯著低于PBS組(P＜0.01)。聯(lián)合抗原組和ESA組小腸沖洗液特異性sIgA均顯著高于PBS組(P＜0.01);PBS組、STAg組、ESA組、聯(lián)合抗原組血清IgG含量依次增高,但無顯著性差異。聯(lián)合抗原組滴鼻免疫小鼠其胚胎感染率均低于其他組。 結(jié)論孕鼠灌胃感染8000個速殖子為建立弓形蟲垂直傳播小鼠模型的適宜劑量。STAg20μg/只滴鼻免疫小鼠(免疫2次,間隔2周),末次免疫后第23天攻蟲可有效降低母鼠感染率及胎盤和胚胎的感染率。STAg聯(lián)合ESA鼻內(nèi)免疫小鼠的效果優(yōu)于單獨STAg或ESA,STAg聯(lián)合ESA(1∶1)20μg/只滴鼻免疫BALB/c小鼠(免疫2次,間隔2周),于末次免疫后第23天攻蟲可降低母鼠感染率及胎盤和胚胎的感染率。
[Abstract]:Objective To establish a mouse model of vertical propagation of Toxoplasma gondii RH strain , and then observe the blocking effect of soluble tachyzoite antigen ( STAg ) on the vertical propagation of Toxoplasma gondii .


Methods This study was divided into three parts : the first part : 54 pregnant BALB / c pregnant mice were randomly divided into 6 groups . On the 8th day of gestation , the BALB / c pregnant mice were infected with 0 , 1000 , 2000 , 4000 , 8000 , 16000 toxoplasmosis tachyzoites , and the live fetuses were sacrificed on the 18th day of gestation , and the toxoplasmosis antigens of the liver , placenta and fetal brain tissues were measured .


In the second part , 130 female BALB / c mice were randomly divided into 2 large groups : PBS group and STAg group . The groups were divided into 1 , 2 and 3 groups with STAg 20渭l / nose and STAg group at 2 : 1 . Each group was divided into 1 , 2 and 3 groups according to the time of immunization .


In the third part , 40 BALB / c virgins were randomly divided into four groups : PBS group , STAg group , ESA group and combined antigen ( STAg + ESA ) group . Each group consisted of 10 . STAg group , ESA group and combined antigen ( STAg + ESA ) group .


Results The mortality of pregnant mice and the infection rate of placenta and embryo increased with the increase of the infective dose , while the rate of live birth was decreased . The rate of embryo infection in 4000 groups was 5.13 % , and the rate of embryo infection in 4000 groups was 5.13 % .


In STAg group 3 , the percentage of fetal thymus weight was significantly higher than that in other groups ( P < 0.05 ) . The percentage of thymus weight of STAg group 3 was higher than that in other groups ( P < 0.05 ) . The infection rate of STAg group 3 was higher than that in other groups ( P < 0.05 ) . The infection rate of STAg group 3 was higher than that in other groups ( P < 0.05 ) .


In STAg group , both ESA group and combined antigen group were significantly lower than that in PBS group ( P < 0.01 ) . There was no significant difference in serum IgG content in STAg group , STAg group , ESA group and combined antigen group ( P < 0.01 ) .


Conclusion The effect of STAg combined ESA intranasal immunization is superior to that of single STAg or ESA , STAg combined ESA ( 1 : 1 ) 20 渭g / mouse immunized BALB / c mice ( 2 times , 2 weeks interval ) . The infection rate of maternal and mouse and the infection rate of placenta and embryo can be reduced at 23 days after the last immunization .
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R392

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本文編號：1994710