MLV-10A1逆转录病毒伪型高效转染人类CD4+T细胞
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本文是一篇研究论文，标题为"MLV-10A1逆转录病毒假型高效转导原始人CD4+ T淋巴细胞"，主要探讨了MLV-10A1逆转录病毒伪型在转导人类CD4+ T淋巴细胞方面的效率。MLV-10A1是一种逆转录病毒，其包膜被改造成与鼠白血病病毒10A1（MLV-10A1）的包膜相似，这种伪型设计被用于提高病毒载体的感染能力。
先前的研究已经证实，相比于其他常见的逆转录病毒包膜，如其他A-MLV（如猴免疫缺陷病毒）、 Gibbon ape leukemia virus (GaLV)和猫内源性逆转录病毒(RD114)，MLV-10A1包膜伪型在转导初级的人类CD8+ T淋巴细胞时表现出更高的效率。考虑到基因疗法（如ADA治疗）和针对HIV的治疗方法的成功依赖于对CD4+ T淋巴细胞的有效转导，这项研究的重点在于评估MLV-10A1能否作为一种有效的工具，用于此类细胞的转化。
研究方法部分详细描述了作者们如何利用编码增强绿色荧光蛋白（EGFP）或其他目的基因的逆转录病毒载体，结合MLV-10A1包膜伪型，来测试其在转导初级人CD4+ T淋巴细胞中的表现。他们可能采用了体外实验，包括病毒包装、感染步骤，并通过检测转染效率、病毒滴度和细胞内的表达水平来评价转导效果。
该研究结果可能具有重要的临床意义，因为它提供了关于如何优化病毒载体设计，以便在治疗策略中更好地利用CD4+ T细胞，这些细胞对于免疫反应和抗病毒治疗至关重要。同时，这项研究也揭示了MLV-10A1包膜在提高病毒对特定细胞类型选择性感染方面的潜力，这对于个性化医疗和靶向治疗的发展具有积极影响。
文章的作者Monika Gladow、Christian Becker、Thomas Blankenstein和Wolfgang Uckert来自德国的Max-Delbrück分子医学中心以及柏林自由大学免疫学研究所，他们的合作有助于推动了逆转录病毒技术在免疫学领域的应用和基础研究进展。最后，该研究在2000年9月18日在线发表，反映了当时对该领域最新研究的理解和进展。
RESEARCH ARTICLE
MLV-10A1 retrovirus pseudotype ef®ciently
transduces primary human CD
4
+
T lymphocytes
Monika Gladow
1
Christian Becker
1
Thomas Blankenstein
1,2
Wolfgang Uckert
1
*
1
Max-Delbru
È
ck-Center for Molecular
Medicine, Berlin, Germany
2
Institute of Immunology, Free
University of Berlin, Germany
*Correspondence to: W. Uckert,
Max-Delbru
È
ck-Center for Molecular
Medicine, Robert-Ro
È
ssle-Strasse 10,
D-13092 Berlin, Germany.
E-mail: wuckert@mdc-berlin.de
Received: 20 June 2000
Revised: 24 July 2000
Accepted: 31 August 2000
Published online: 18 September 2000
Abstract
Background Previously, we showed that retroviral vectors pseudotyped
with the envelope of the amphotropic murine leukemia virus 10A1 (MLV-
10A1) more ef®ciently transduce primary human CD8
+
T lymphocytes when
compared with other A-MLV, gibbon ape leukemia virus (GaLV) and feline
endogenous retrovirus (RD114) vector pseudotypes. For the success of
several gene therapeutic approaches (ADA, HIV) it is important to effectively
transduce primary human CD4
+
T lymphocytes.
Methods We have used retroviral vectors encoding the enhanced green
¯uorescent protein (EGFP) as a marker gene and carrying envelopes of MLV-
10A1, A-MLV and GaLV and have analyzed the transduction ef®ciency of both
human CD4
+
T cell lines (CEM, H9, HUT78, J16) and primary human CD4
+
T lymphocytes using a RetroNectin-assisted transduction protocol and virus-
containing supernatant.
Results In CD4
+
T cell lines the MLV-10A1 vector pseudotype was most
effective and infected up to 85% of cells which then stably expressed GFP over
time. MLV-10A1 was also superior and infected approximately 32% of
primary human CD4
+
T lymphocytes in comparison to GaLV (18%) and A-
MLV (12%). The superior ef®ciency of MLV-10A1 for the transduction of
CD4
+
T cells correlates with the longer half-life of this pseudotype in
comparison to A-MLV and, as previously shown by interference analysis, with
the usage of both the A-MLV (Pit2) and the GaLV receptor (Pit1) for cell entry.
Conclusions MLV-10A1 is a suitable vector for transferring genes with high
ef®cacy into primary human CD4
+
T lymphocytes. The use of MLV-10A1
pseudotyped vectors should make it easier to obtain a suf®cient number of
gene-modi®ed T lymphocytes for an adoptive transfer. Copyright # 2000
John Wiley & Sons, Ltd.
Keywords Gene transfer; T cell transduction; MLV-10A1 retrovirus vector
pseudotype; GFP marker gene
Introduction
The adoptive transfer of genetically modi®ed T lymphocytes is a promising
approach for immunotherapy of cancer as well as for genetic and infectious
diseases. The safety and feasibility of administering gene-modi®ed T cells to
patients has been demonstrated in clinical trials using, e.g., cytokine gene-
modi®ed tumor-in®ltrating T lymphocytes (TIL) to enhance anti-tumor
activity [1], autologous CD4
+
T lymphocytes encoding the adenosine
deaminase (ADA) gene to reconstitute the immune function of ADA-SCID
patients [2±4], CD4
+
T lymphocytes to inhibit HIV expression and replication
THE JOURNAL OF GENE MEDICINE
J Gene Med 2000; 2: 409±415.
Copyright # 2000 John Wiley & Sons, Ltd.
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