發(fā)布時(shí)間：2018-01-12 16:44

  本文關(guān)鍵詞：北平頂猴血清免疫學(xué)指標(biāo)檢測及其APOBEC3家族基因克隆和抗HIV-1研究　出處：《昆明理工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 北平頂猴 動物模型 免疫球蛋白 補(bǔ)體 APOBEC3 HIV

【摘要】：隨著生物醫(yī)學(xué)研究的深入,靈長類動物模型在醫(yī)學(xué)研究領(lǐng)域的重要性愈發(fā)明顯。在研究艾滋病、麻疹、瘧疾、肝炎和神經(jīng)退行性疾病等重大人類疾病以及認(rèn)知等基礎(chǔ)研究方面,靈長類動物是首選或者唯一的實(shí)驗(yàn)動物,其研究成果更容易轉(zhuǎn)化為臨床應(yīng)用,可以大大降低新藥研發(fā)的風(fēng)險(xiǎn)。與其他非人靈長類動物相比,獼猴屬的平頂猴(pig-tailed macaques, Macaca nemestrina group, PTM)、恒河猴(rhesus macaques, Macaca mulatta, RM)與食蟹猴(cynomolgus macaques, Macaca fascicularis, CM)在人類生物醫(yī)學(xué)研究領(lǐng)域應(yīng)用最為廣泛,其中平頂猴是唯一對HIV-1易感的舊大陸猴,被廣泛用于HIV-1的致病機(jī)制、藥物評價(jià)及疫苗等AIDS研究領(lǐng)域,此外平頂猴在其它感染性疾病、神經(jīng)生理及行為認(rèn)知等研究領(lǐng)域也得到了廣泛應(yīng)用。目前,平頂猴被分為3個(gè)種：南平頂猴(Southern pig-tailed macaque, M. nemestrina)、北平頂猴(Northern pig-tailed macaque, M. leonina, NPM)和明打威猴(Mentawai macaque, M. pagensis)。了解北平頂猴的基礎(chǔ)生物學(xué)數(shù)據(jù),不僅對于北平頂猴健康狀態(tài)的判定、疾病的發(fā)生與發(fā)展、疾病的治療具有重要的參考價(jià)值,而且對于北平頂猴的飼養(yǎng)管理和疾病模型研究也具有重要的指導(dǎo)意義。盡管平頂猴能夠感染HIV-1,但是其在平頂猴體內(nèi)的復(fù)制水平受到明顯抑制,這極大地限制了HIV-1感染平頂猴模型在艾滋病疫苗、藥物篩選及致病機(jī)制等研究領(lǐng)域的應(yīng)用。感染HIV-1的平頂猴體內(nèi)的HIV-1病毒存在大量的G到A突變,其突變頻率遠(yuǎn)遠(yuǎn)超過了自發(fā)突變的頻率,提示APOBEC3家族成員在限制HIV-1在平頂猴體內(nèi)的復(fù)制方面也許發(fā)揮了重要的作用。我們之前報(bào)道過有關(guān)北平頂猴血液學(xué)和血液生化指標(biāo)參考值,但是關(guān)于北平猴血清免疫球蛋白,補(bǔ)體與C反應(yīng)蛋白(CRP)等免疫學(xué)常規(guī)指標(biāo)的基礎(chǔ)數(shù)據(jù)還未見國內(nèi)外報(bào)道。本研究對不同性別與年齡段健康實(shí)驗(yàn)北平頂猴的血清IgA、IgG、IgM、 C3、C4及CRP進(jìn)行了測定與分析,結(jié)果表明人工飼養(yǎng)的北平頂猴血清IgA、IgM、 C3、C4水平不隨年齡發(fā)生變化,而血清IgG水平隨年齡增長有增加的趨勢(r=0.202；P=0.045)。血清IgG、IgA、IgM、C3水平在雌性中明顯高于雄性(P0.05),而血清C4水平在雌雄間差異不顯著(P=0.777)。此外,我們發(fā)現(xiàn)血清C3與C4水平間有很強(qiáng)的正相關(guān)性(r=0.700；P0.0001)。該研究為北平頂猴的飼養(yǎng)與生物醫(yī)學(xué)研究提供了一些較重要的血清免疫球蛋白、補(bǔ)體的基礎(chǔ)數(shù)據(jù),北平頂猴免疫學(xué)相關(guān)數(shù)據(jù)的進(jìn)一步檢測和完善將會促進(jìn)北平頂猴在感染性疾病、免疫學(xué)等生物醫(yī)學(xué)研究領(lǐng)域的有效應(yīng)用。此外,本研究成功克隆了北平頂猴APOBEC3A-APOBEC3H基因,序列比對及核酸一致性分析發(fā)現(xiàn)北平頂猴的APOBEC3A-APOBEC3H基因蛋白編碼區(qū)序列與恒河猴、食蟹猴、南平頂猴及人的相應(yīng)基因的序列的一致性高達(dá)890%～99%,尤其是APOBEC3G與APOBEC3F基因其蛋白編碼區(qū)的核酸序列與恒河猴及南平頂猴相應(yīng)基因序列的一致性高達(dá)99%。另外,本研究對北平頂猴APOBEC3A-APOBEC3H基因的體外抗HIV-1活性進(jìn)行了研究,結(jié)果顯示北平頂猴APOBEC3G/F的抗HIV-1的活性最強(qiáng),APOBEC3B/D抗HIV-1的活性次之,APOBEC3H的抗HIV-1活性比APOBEC3B/D弱,APOBEC3A/C沒有表現(xiàn)出抗HIV-1的活性。本研究提示APOBEC3家族中APOBEC3G與APOBEC3F也許是限制HIV-1在平頂猴體內(nèi)復(fù)制的重要宿主限制因子之一,對HIV-1感染的平頂猴模型的優(yōu)化提供了有參考價(jià)值的信息。
[Abstract]:Along with the development of biomedical research, the primate animal model of importance in the field of medical research is becoming increasingly apparent. In the study of AIDS, measles, malaria, hepatitis and basic research of neurodegenerative diseases and other major human diseases and cognition, primate animal or animal is preferred only, the results are more easily transformed into clinical application, can greatly reduce the risk of drug development. Compared with other non-human primate animal, Macaca monkfy (pig-tailed macaques, the Macaca nemestrina group, PTM), Monkey (rhesus macaques, Macaca Ganges RIver mulatta, RM) and cynomolgus monkeys (cynomolgus macaques, Macaca fascicularis, CM) in the field of Applied Research in the biomedical human widely, which is the only monkfy susceptible to HIV-1 the Old World monkey, is widely used in the pathogenic mechanism of HIV-1 and AIDS on drug evaluation and vaccine In addition, monkfy in other infectious diseases, neurophysiological and cognitive behavioral research has been widely used. At present, monkfy are divided into 3 types: Nanping top Monkey (Southern pig-tailed macaque, M. nemestrina) (Northern pig-tailed, Beijing top monkey macaque, M. leonina, NPM) and the Mentawai macaque (Mentawai macaque, M. pagensis). To understand the basic biological data of Peking monkey, not only to determine the health status of the Peking monkey, the occurrence and development of disease, which has important reference value in the treatment of disease, but also has an important guiding significance for the study of feeding management and disease model of Peking monkey. Although monkfy to HIV-1 infection, but the replication level monkfy in vivo was inhibited, which greatly limits the HIV-1 infection monkfy model in AIDS vaccine, drug screening and pathogenic mechanism Study on the application of HIV-1. Flat monkeys infected with the HIV-1 virus, there are a large number of G to A mutation, the mutation frequency is far more than the frequency of spontaneous mutations, suggesting that APOBEC3 family members in the limit of HIV-1 in monkfy replication in vivo may play an important role. We have previously reported the Peking monkey blood and the blood biochemical indexes of the reference value, but about Beijing monkey serum immunoglobulin, complement C and C-reactive protein (CRP) based data there is no conventional indicators of immunology reported at home and abroad. The levels of IgA, the research on the Beijing top monkey with different gender and age healthy experiment of IgG, IgM, C3, C4 and CRP were measured and analyzed. The results show that the artificial feeding Peking monkey serum IgA, IgM, C3, C4 levels do not change with age, and the serum IgG levels with age increasing (r=0.202; P =0.045). Serum IgG, IgA, IgM, C3 levels in females was significantly higher than that of male (P0.05), and the serum C4 levels in female and male with no significant difference (P=0.777). In addition, we found that serum C3 and C4 level, there is a strong positive correlation between (r=0.700; P0.0001). This study provided some serum immunoglobulin is more important as Peking monkey breeding and bio medical research, basic data complement, Beijing top further detection of relevant data and improve the monkey will promote the immunological Peking monkey in infectious diseases, the effective application of immunology and other biomedical research field. In addition, this study successfully cloned Peking monkey APOBEC3A-APOBEC3H gene, nucleic acid sequence alignment and consensus analysis found that the encoding sequence of APOBEC3A-APOBEC3H protein and the Ganges RIver Peking monkey monkey, Macaca fascicularis, consensus sequences corresponding genes in Nanping and the top of the monkey is up to 890% to 99%, especially APOBE The consistency of C3G nucleotide sequence of APOBEC3F gene and its protein encoding region and Ganges RIver and Nanping top the corresponding gene sequence of monkey monkey up to 99%. in the study of Peking monkey APOBEC3A-APOBEC3H gene in vitro anti HIV-1 activity were studied, the results showed that the anti HIV-1 activity of the strongest Peking monkey APOBEC3G/F, APOBEC3B/D anti HIV-1 activity second, APOBEC3H anti HIV-1 activity was weaker than APOBEC3B/D, APOBEC3A/C showed no anti HIV-1 activity. This study suggests that the APOBEC3 family APOBEC3G and APOBEC3F may be one of the main limiting factor in limiting HIV-1 important places monkfy replication, provides valuable information to optimize monkfy model of HIV-1 infection.

【學(xué)位授予單位】：昆明理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.6;R512.91

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李致祥,林正玉;云南靈長類的分類和分布[J];動物學(xué)研究;1983年02期

2 朱林;張高紅;鄭永唐;;動物模型在HIV-1殺微生物劑有效性和安全性評價(jià)中的應(yīng)用[J];動物學(xué)研究;2010年01期

3 夏厚軍;陳策實(shí);;非人靈長類腫瘤模型研究進(jìn)展(英文)[J];動物學(xué)研究;2011年01期

4 唐向輝;曹躍齡;楊澤星;趙富鮮;;多囊卵巢綜合征獼猴的生殖特性(英文)[J];動物學(xué)研究;2012年01期

5 張艷春;靳麗莎;高麗華;彭白露;何紅;季芳;郝香芬;張秀娟;唐湘北;饒軍華;劉曉明;;高能量膳食誘導(dǎo)食蟹猴2型糖尿病模型的研究[J];動物醫(yī)學(xué)進(jìn)展;2012年08期

6 徐林;張?jiān)?梁斌;呂龍寶;陳策實(shí);陳勇彬;周巨民;姚永剛;;實(shí)驗(yàn)動物樹，

本文編號：1415122