Establishment of an in vitro growth competition assay for HIV-1 CRF07_BC viral fitness
10.3760/cma.j.cn112866-20210112-00009
- VernacularTitle:HIV-1 CRF07_BC病毒适应性的体外生长竞争试验的建立
- Author:
Lin HE
1
;
Yang JIAO
;
Yibo DING
;
Yuanyuan ZHANG
;
Qianru LIN
;
Lijie WANG
;
Dan LI
;
Kunxue HONG
;
Liying MA
Author Information
1. 中国疾病预防控制中心性病艾滋病预防控制中心，北京 102206
- Keywords:
HIV-1;
CRF07_BC;
Vitro growth competition assay;
Viral fitness
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(4):457-462
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish an in vitro growth competition assay using HIV-1 CRF07_BC infectious clone containing the mouse Thy1.1 gene or Thy1.2 gene. Methods:The mouse Thy1.1 gene and Thy1.2 gene were used to replace the 218 bases of Nef region of the CRF07_BC infectious clone (pXJDC6291-13) and construct the CRF07_BC infectious clones BCEA1 and BCEA2 plasmid. BCEA1 and BCEA2 plasmids were transfected into 293T to obtain the virus and the viral titration was measured. PM1 cells were co-infected with an equal virus. On the 3 rd to 6 th day of infection the cells were collected and labeled with specific antibodies Thy1.1 and Thy1.2. The viral fitness was detected by flow cytometry. Through "TFitness" (http: //bis.urmc.rochester.edu/vFitness) the relative viral fitness was calculated. The influence K103 N and K166R on the relative fitness of HIV-1 CRF07_BC were observed. Results:The relative fitness (1+ s) of CRF07_BC subtypes BCEA1 and BCEA2 viruses were 1.06±0.23693, which was not statistically significant. An in vitro growth competition assay for CRF07_BC was established. The in vitro growth competition assay was used to verify the relative fitness of the K103 N and K166R mutant strains to the wild strain. The relative fitness of BCEA2-K103 N/BCEA1 was 0.728282±0.16608, BCEA2-K166R/BCEA1 was 0.883385±0.19023, BCEA2-K103 N+ K166R/BCEA1 was 0.804604±0.06164. The relative fitness of K103 N resistant mutant strain and the wild strain was significantly different.Conclusions:An in vitro growth competition assay for the fitness of HIV-1 CRF07_BC virus was established. Based on the in vitro growth competition assay, the HIV-1 K103 N resistant mutant strain has reduced viral fitness.
