2012年8月17日 訊 /生物谷BIOON/ --盡管瘧疾和HIV都是世界性大流行疾病,，而且同時(shí)感染這兩種疾病能夠?qū)е卤容^高的死亡率，但是科學(xué)家們對(duì)HIV-1和導(dǎo)致瘧疾的惡性瘧原蟲(chóng)(Plasmodium falciparum)之間的相互作用仍然知之甚少,。然而,，根據(jù)一篇刊登在Journal of Visualized Experiments(JoVE)期刊上的視頻論文，研究人員描述了一種新技術(shù)來(lái)在體外培養(yǎng)的人細(xì)胞中研究HIV-1和惡性瘧原蟲(chóng)之間的相互作用,，以便允許科學(xué)家們研究這兩種微生物共同感染宿主細(xì)胞的不同參數(shù),。
領(lǐng)導(dǎo)這項(xiàng)研究的David Richard博士解釋道，“被HIV-1感染的免疫細(xì)胞遭遇這種瘧原蟲(chóng)時(shí)會(huì)發(fā)生什么,，我們還不太了解,。來(lái)自幾項(xiàng)探究這兩種疾病相互作用的研究結(jié)果有時(shí)是相互沖突的。我們希望我們的方法將允許我們?cè)谝粋€(gè)簡(jiǎn)化的系統(tǒng)中充分地研究這些相互作用,。”
在這兩種疾病中,，每種疾病都攻擊人血液中不同的組分，因而破壞正常的免疫功能,。惡性瘧原蟲(chóng)感染紅細(xì)胞導(dǎo)致病人發(fā)熱,、顫抖、嘔吐或驚厥,。HIV-1通過(guò)感染包括巨噬細(xì)胞和輔助性T細(xì)胞在內(nèi)的免疫系統(tǒng)組分,，然后進(jìn)行自我復(fù)制和破壞宿主細(xì)胞，從而導(dǎo)致病人患上獲得性免疫缺陷綜合癥(acquired immune deficiency syndrome, AIDS),。通過(guò)在體外研究每種疾病不同階段的共同感染情形,，科學(xué)家們能夠更好地理解瘧疾感染和HIV復(fù)制的不同階段如何影響另一種疾病的發(fā)生和嚴(yán)重性。為此,，Richard博士和他的實(shí)驗(yàn)室呈現(xiàn)一種技術(shù)來(lái)研究被惡性瘧原蟲(chóng)感染的紅細(xì)胞如何影響HIV-1在單核細(xì)胞源性巨噬細(xì)胞(monocyte-derived macrophage)中的復(fù)制,。
Richard博士指出，“通過(guò)這篇發(fā)表在JoVE期刊上的論文,，人們能夠在實(shí)驗(yàn)中觀察到發(fā)生什么,。這項(xiàng)視頻展示有助于簡(jiǎn)單地理解一段較長(zhǎng)的實(shí)驗(yàn)程序和在細(xì)胞水平上更加充分地描繪出這些相互作用的圖譜。”他希望這篇論文的發(fā)表將讓科學(xué)界能夠在細(xì)胞水平上研究這些相互作用,，這將是改善感染上這些疾病的個(gè)人的生活質(zhì)量的第一步,。JoVE期刊編輯Charlotte Frank Sage博士陳述道，“這種實(shí)驗(yàn)程序提供一種工具來(lái)研究惡性瘧原蟲(chóng)和HIV之間的相互作用,。JoVE期刊發(fā)表的這種實(shí)驗(yàn)程序?qū)⒃试S全世界的科學(xué)家們能夠詳細(xì)地驗(yàn)證這種系統(tǒng),，從而有助于將這種技術(shù)應(yīng)用到他們的實(shí)驗(yàn)室之中。” (生物谷Bioon.com)
本文編譯自Novel technique demonstrates interactions between malaria parasite and HIV
doi: 10.3791/4166
PMC:
PMID:
An In vitro Co-infection Model to Study Plasmodium falciparum-HIV-1 Interactions in Human Primary Monocyte-derived Immune Cells
Guadalupe Andreani, Dominic Gagnon, Robert Lodge, Michel J. Tremblay, Dave Richard
Plasmodium falciparum, the causative agent of the deadliest form of malaria, and human immunodeficiency virus type-1 (HIV-1) are among the most important health problems worldwide, being responsible for a total of 4 million deaths annually1. Due to their extensive overlap in developing regions, especially Sub-Saharan Africa, co-infections with malaria and HIV-1 are common, but the interplay between the two diseases is poorly understood. Epidemiological reports have suggested that malarial infection transiently enhances HIV-1 replication and increases HIV-1 viral load in co-infected individuals2,3. Because this viremia stays high for several weeks after treatment with antimalarials, this phenomenon could have an impact on disease progression and transmission.
The cellular immunological mechanisms behind these observations have been studied only scarcely. The few in vitro studies investigating the impact of malaria on HIV -1 have demonstrated that exposure to soluble malarial antigens can increase HIV-1 infection and reactivation in immune cells. However, these studies used whole cell extracts of P. falciparum schizont stage parasites and peripheral blood mononuclear cells (PBMC), making it hard to decipher which malarial component(s) was responsible for the observed effects and what the target host cells were4,5. Recent work has demonstrated that exposure of immature monocyte-derived dendritic cells to the malarial pigment hemozoin increased their ability to transfer HIV-1 to CD4+ T cells6,7, but that it decreased HIV-1 infection of macrophages8. To shed light on this complex process, a systematic analysis of the interactions between the malaria parasite and HIV-1 in different relevant human primary cell populations is critically needed.
Several techniques for investigating the impact of HIV-1 on the phagocytosis of micro-organisms and the effect of such pathogens on HIV-1 replication have been described. We here present a method to investigate the effects of P. falciparum-infected erythrocytes on the replication of HIV-1 in human primary monocyte-derived macrophages. The impact of parasite exposure on HIV-1 transcriptional/translational events is monitored by using single cycle pseudotyped viruses in which a luciferase reporter gene has replaced the Env gene while the effect on the quantity of virus released by the infected macrophages is determined by measuring the HIV-1 capsid protein p24 by ELISA in cell supernatants.
