Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Single-round infection assay

JK Joseph Kononchik
JI Joanna Ireland
ZZ Zhongcheng Zou
JS Jason Segura
GH Genevieve Holzapfel
AC Ashley Chastain
RW Ruipeng Wang
MS Matthew Spencer
BH Biao He
NS Nicole Stutzman
DK Daiji Kano
JA James Arthos
EF Elizabeth Fischer
TC Tae-Wook Chun
SM Susan Moir
PS Peter Sun
request Request a Protocol
ask Ask a question
Favorite

Stimulated CD8− PBMC were resuspended at 2×106/mL in culture media. Aliquots of 200 µL (4×105 cells) were transferred to 96-well plates for incubation in triplicate with anti-CD4 (10 µg/mL), anti-CD62L (30 µg/mL), or isotype (30 µg/mL) antibody at 37 °C for 60 min prior to the addition of virus. Luciferase viruses pseudotyped with envelopes from JRFL (R5-tropic) and SF33 (X4-tropic) HIV-1 and VSV were added to the cells at a concentration of 100 ng/mL HIV p24. The infected CD8− PBMC were then incubated for 72 h, lysed, and assayed for luciferase activity according to the manufacturer’s recommendations (Promega Corporation, Madison, WI). Pseudotyped virus from GNTI− cells were added at 100 ng/mL to cells as above.

Copyright and License information: The Author(s) ©2018
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A