Virologica Sinica

, Volume 33, Issue 2, pp 142–152 | Cite as

Differential Effects of Strategies to Improve the Transduction Efficiency of Lentiviral Vector that Conveys an Anti-HIV Protein, Nullbasic, in Human T Cells

  • Lina Rustanti
  • Hongping Jin
  • Dongsheng Li
  • Mary Lor
  • Haran Sivakumaran
  • David HarrichEmail author
Research Article


Nullbasic is a mutant form of HIV-1 Tat that has strong ability to protect cells from HIV-1 replication by inhibiting three different steps of viral replication: reverse transcription, Rev export of viral mRNA from the nucleus to the cytoplasm and transcription of viral mRNA by RNA polymerase II. We previously showed that Nullbasic inhibits transduction of human cells including T cells by HIV-1-based lentiviral vectors. Here we investigated whether the Nullbasic antagonists huTat2 (a Tat targeting intrabody), HIV-1 Tat or Rev proteins or cellular DDX1 protein could improve transduction by a HIV-1 lentiviral vector conveying Nullbasic-ZsGreen1 to human T cells. We show that overexpression of huTat2, Tat-FLAG and DDX1-HA in virus-like particle (VLP) producer cells significantly improved transduction efficiency of VLPs that convey Nullbasic in Jurkat cells. Specifically, co-expression of Tat-FLAG and DDX1-HA in the VLP producer cell improved transduction efficiency better than if used individually. Transduction efficiencies could be further improved by including a spinoculation step. However, the same optimised protocol and using the same VLPs failed to transduce primary human CD4+ T cells. The results imply that the effects of Nullbasic on VLPs on early HIV-1 replication are robust in human CD4+ T cells. Given this significant block to lentiviral vector transduction by Nullbasic in primary CD4+ T cells, our data indicate that gammaretroviral, but not lentiviral, vectors are suitable for delivering Nullbasic to primary human T cells.


HIV-1 Transduction Lentiviral vector Nullbasic Tat Rev DDX1 Intrabody 



This research was supported by the National Health and Medical Research Council Project Grant (1085359). LR was supported by Prime Minister’s Australia Asia Endeavour Postgraduate (Ph.D.) Award funded by the Australian Government, Department of Education and Training, UQ international scholarship (UQI) and UQ Centenial scholarship (UQCent). We thank the QIMR Berghofer Flow Cytometry and Imaging Facility for technical expertise with cell sorting and flow cytometry. We thank Wayne Marasco for providing the sFvhutat2 expression plasmid for this study.

Author Contributions

DH and LR originated and guided the study. DL, DH, LR and HJ designed the experiments. LR, ML and HJ performed experiments. LR, DH, HJ and DL wrote the manuscript. All authors read and approved the final manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

“Buffy coat” human blood cells were obtained from the Australian Red Cross. All samples were provided by donors with informed consent.


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Copyright information

© Wuhan Institute of Virology, CAS and Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  1. 1.Faculty of MedicineThe University of QueenslandHerstonAustralia
  2. 2.Department of Cell and Molecular BiologyQIMR Berghofer Medical Research InstituteHerstonAustralia
  3. 3.National Institute of Health Research and Developmentthe Ministry of Health of Republic of IndonesiaCentral JakartaIndonesia

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