Retroviral vector - mediated expression of human interleukin-4 gene in human tumor cells

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Retroviral vector - mediated expression of human interleukin-4 gene in human tumor cells

更新时间：2024-01-17

- Retroviral vector - mediated expression of human interleukin-4 gene in human tumor cells
- Journal of Chongqing Medical University Issue 1, (1998)
- 作者机构：
  
  儿科学院免疫室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R733.4;R733.7
- Published：1998，
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[1]赵晓东,李成荣,杨锡强,蒋丽萍.人IL-4重组逆转录病毒载体的构建及在人肿瘤细胞中的表达[J].重庆医科大学学报,1998(01):1-4.

Retroviral vector - mediated expression of human interleukin-4 gene in human tumor cells. [J]. Journal of Chongqing Medical University (1).(1998)
[1]赵晓东,李成荣,杨锡强,蒋丽萍.人IL-4重组逆转录病毒载体的构建及在人肿瘤细胞中的表达[J].重庆医科大学学报,1998(01):1-4. DOI：

Retroviral vector - mediated expression of human interleukin-4 gene in human tumor cells. [J]. Journal of Chongqing Medical University (1).(1998) DOI：

摘要

利用多聚酶链反应（ＰＣＲ）方法，从ＰＣＤ－ｈＩＬ－４质粒中扩增得到人白细胞介素４（ｈＩＬ－４）的ｃＤＮＡ－。ＤＮＡ序列测定证实此片段包含完整的ＩＬ－４开放阅读框架。应用ＤＮＡ重组技术，将此ｃＤＮＡ重组于逆转录病毒载休ＬＸＳＮ。用脂质体转染法将此重组质粒导入病毒包装细胞ＰＡ３１７。得到滴度为２．５ｘ１０ＣＦＵ／ｍｌ的感染性病毒。病毒感染人白血病细胞株ＨＬ－６０、Ｋ５６２及Ｂｕｒｋｉｔ淋巴瘤细胞株Ｒａｊｉ，使之表达并分泌ＩＬ－４。逆转录ＰＣＲ（ＲＴ－ＰＣＲ）法证实ｈＩＬ－４ｃＵＮＡ可在上述肿瘤细胞中持续转录。ＥＬＩＳＡ怯证实病肿瘤细胞分辨ＩＬ一４水平可高达３００Ｐ／１０细胞．２４小时。本研究为进一步探讨转ＩＬ一４基因用于血液系统恶性肿瘤免疫治疗的价值提供了基础。

Abstract

Human interleukin - 4 complementory DNA (hlL - cDNA) was amplified from a hlL -4 cDNA - positive plasmid

PCD - hIL - 4 by PCR. Sequence analysis showed no mutation occured and the open reading frame was included in the fragmcnt. Human hIL - 4 cDNA was inserted into polylinker site of retroviral vector

LXSN to construct recombinant retroviral vector LXSN / hlL - 4

in which the transcription of hlL - 4 cDNA drived by 5’LTR promotcr. The recombinant retrovirals was pakaged in PA317 cell line and used to infect human HL - 60

k 562 and Raji cell linc in presence of 4ug/ml polybrene. retroJranscriptimal PCR analysis abowed hlL - 4 cDNA was trancripted in modified tumor cells and the maxrnium of IL-4 level secreted by modified tumor cells obtained 300pg/10 cell - 24h.The results suggested that hlL - 4 cDNA could be expressed efficiently under the control of vector’s promoter.

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IL－4基因转移基因表达

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