- A cutting edge technology in HIV research
HIV InfectinTM is designed to facilitate HIV penetration of the cortical actin barrier, thereby greatly facilitating productive viral infection. HIV Infectin can be used to enhance the infection of a variety of host cells by HIV (both X4 and R5 viruses) to facilitate biochemical and biological characterization of HIV infection. HIV Infectin can enhance HIV infection rate 5 to 30 fold. Virongy developed HIV Infectin is based on the scientific theory that the actin cytoskeleton is a natural barrier for HIV entry and intracellular migration (Yoder et al., Cell, 2008, 134:782).
- Once thawed, the HIV Infectin should be stored at 4oC, and is stable for 3 months.
- Do not re-freeze and do not leave HIV Infectin at room temperature.
- HIV Infectin is an HIV-specific infectivity enhancer and works with any HIV-receptive cell line to enhance HIV infectivity rates.
- HIV Infectin increases productive viral infection by 5 to 30 fold.
Use with the HIV Rev-dependent reporter cells and any other HIV target cells:
- Anti-HIV drug screenings via one-step infection
- Biochemistry characterization (DNA, RNA, protein) of HIV-infected cells
- HIV viral protein interaction with viral and host cell proteins
- Screenings for broadly neutralizing antibodies (bnAB) (from laboratory and clinical research samples)
- Neutralizing antibody quantifications
- HIV cell-cell transmission and HIV drug-resistance studies
- HIV host restriction factor (HRF) studies
- HIV host dependency factor (HDF) studies
- Low-level HIV gene expression assessments
- HIV pre-integration transcription studies
- HIV latency and reactivation studies
- HIV outgrowth detection following reactivation
- HIV tropism determinations
Table 1. HIV Infectin is formulated at 10X concentration
Protocol - HIV Infection of CD4 T cells
(see Table 2 for scale-up recommendations)
1. Count cells to be infected, pellet cells by centrifugation at 300 x g for 5 minutes.
Note: Cell viability should be ≥ 80%.
2. Resuspend cells in complete media at a concentration of ~2 x 106 cells ml-1.
3. Use 100 μl of cells (~2 x 105 cells) per infection.
4. Pre-treat cells by adding 10 μl of HIV Infectin so that the HIV Infectin concentration is 1X. Mix and incubate for 2 hours.
For best results, use HIV Infectin with HIV Rev-dependent Reporter cells.
5. Add virus to cells and mix. Note volume of virus used.
6. Add HIV Infectin to 1/10 of the virus volume used, e.g., if 100 μl of virus used, add 10 μl of HIV Infectin. Incubate at 37°C for 2–4 hours.
7. Wash cells by adding an additional 1 ml fresh complete media, pelleting cells as above, and removing supernatant. (Optional): Repeat once for a total of two washes.
8. After washing, resuspend cells in 1 ml complete medium.
9. Culture and utilize cells as usual.
Table 2: Scale-up recommendations for HIV infection using HIV InfectinTM
Yoder A, Yu D, Dong L, Iyer SR, Xu X, Kelly J, et al. HIV envelope-CXCR4 signaling activates cofilin to overcome cortical actin restriction in resting CD4 T cells. Cell. 2008; 134(5):782-92.
HIV Infectin is intended for research use only and is not for diagnostic or therapeutic purposes or uses in humans or animals.