CD45 immunoaffinity depletion of vesicles from Jurkat T cells demonstrates that exosomes contain CD45: no evidence for a distinct exosome/HIV-1 budding pathway.
CD45 immunoaffinity depletion of vesicles from Jurkat T cells demonstrates that exosomes contain CD45: no evidence for a distinct exosome/HIV-1 budding pathway.
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DOI：
10.1186/1742-4690-5-64
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发表时间：
2008-07-16
期刊：
影响因子：
3.3
通讯作者：
Ott, David E.
中科院分区：
文献类型：
--
作者：
Coren, Lori V.;Shatzer, Teresa;Ott, David E.
在密度纯化病毒粒子制剂中存在相对高水平的细胞蛋白污染是造血细胞产生的HIV和SIV生化分析中的一个混淆因素。这种污染的主要来源是囊泡，无论是微囊泡还是外泌体，它们具有与病毒粒子相似的物理特性。因此，这些颗粒不能通过大小或密度分馏去除。尽管病毒粒子和囊泡具有相似的细胞蛋白组成，但CD45在HIV-1中被排除在外，但在造血细胞产生的囊泡中却存在。利用这一发现，我们开发了一种CD45免疫亲和去除程序，从HIV-1制剂中去除囊泡。虽然这种方法已经成功地应用于几种不同细胞类型的病毒粒子制备，但一些研究小组已经得出结论，来自某些T细胞系的“外泌体”，特别是Jurkat，不含CD45。如果这种解释是正确的，那么这些囊泡不能通过CD45免疫亲和耗尽来去除。在这里，我们发现Jurkat和SupT1/CCR5细胞产生的致密囊泡含有CD45，并通过CD45免疫亲和耗尽有效地从制剂中去除。此外，从这些细胞系生产的病毒粒子制剂中去除污染的细胞蛋白。以前，“外泌体”和病毒粒子中CD45的缺失被用来支持所谓的特洛伊外泌体假说，即HIV-1只是一个含有病毒物质的外泌体。CD45在囊泡（包括外泌体）上的存在和在病毒粒子上的缺失反对外泌体和HIV-1共有的特殊出芽途径。
The presence of relatively high levels of cellular protein contamination in density-purified virion preparations is a confounding factor in biochemical analyses of HIV and SIV produced from hematopoietic cells. A major source of this contamination is from vesicles, either microvesicles or exosomes, that have similar physical properties as virions. Thus, these particles can not be removed by size or density fractionation. Although virions and vesicles have similar cellular protein compositions, CD45 is excluded from HIV-1 yet is present in vesicles produced from hematopoietic cells. By exploiting this finding, we have developed a CD45 immunoaffinity depletion procedure that removes vesicles from HIV-1 preparations. While this approach has been successfully applied to virion preparations from several different cell types, some groups have concluded that "exosomes" from certain T cell lines, specifically Jurkat, do not contain CD45. If this interpretation is correct, then these vesicles could not be removed by CD45 immunoaffinity depletion. Here we show that dense vesicles produced by Jurkat and SupT1/CCR5 cells contain CD45 and are efficiently removed from preparations by CD45-immunoaffinity depletion. Also, contaminating cellular proteins were removed from virion preparations produced by these lines. Previously, the absence of CD45 from both "exosomes" and virions has been used to support the so called Trojan exosome hypothesis, namely that HIV-1 is simply an exosome containing viral material. The presence of CD45 on vesicles, including exosomes, and its absence on virions argues against a specialized budding pathway that is shared by both exosomes and HIV-1.
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影响因子：
5.4
作者：
Ott, DE;Coren, LV;Henderson, LE
通讯作者：
Henderson, LE
DOI：
10.1073/pnas.72.8.2895
发表时间：
1975-01-01
影响因子：
11.1
作者：
POLISKY, B;MCCARTHY, B
通讯作者：
MCCARTHY, B
影响因子：
4.8
作者：
Wubbolts, R;Leckie, RS;Stoorvogel, W
通讯作者：
Stoorvogel, W
DOI：
10.1083/jcb.200508014
发表时间：
2006-03-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Booth AM;Fang Y;Fallon JK;Yang JM;Hildreth JE;Gould SJ
通讯作者：
Gould SJ
DOI：
10.1083/jcb.101.3.942
发表时间：
1985-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Pan BT;Teng K;Wu C;Adam M;Johnstone RM
通讯作者：
Johnstone RM