Engineering T-Cell Resistance to HIV-1 Infection via Knock-In of Peptides from the Heptad Repeat 2 Domain of gp41

Abstract

HIV is a human lentivirus that infects CD4-positive immune cells and, when left untreated, manifests in the fatal disease known as AIDS. Antiretroviral therapy (ART) does not leading to viral clearance, and HIV persists in the organism as a latent provirus.

Ultrasensitive quantification of HIV-1 cell-to-cell transmission in primary human CD4 + T cells measures viral sensitivity to broadly neutralizing antibodies

“…For these experiments, we generated an additional pUCHR-based vector, designated pUCHR-EF1a-inNluc, in which we replaced the CMV promoter with EF1a-HTLV-1 hybrid promotor to study the effect of promoter on the measurements in primary cells. We isolated CD4 + T cells from two donors, and activated and then electroporated the cells as previously described ( 29 ). All subsequent steps were performed as described for the T cell lines, but transfected cells were cocultured with autologous primary CD4 + T cells.…”

Application of CRISPR/Cas Genomic Editing Tools for HIV Therapy: Toward Precise Modifications and Multilevel Protection

“…A combination of secreted and GPI-anchored forms of MT-C34 via peptide concatemeric construct knocked-in into CXCR4 gene will provide the selection of CD4 + T cells that are resistant to HIV and secrete the MT-C34 peptide which will shield non-edited cells from HIV infection. The GPI-anchored C-peptide KI concept was described previously in our paper ( Maslennikova et al., 2022 ) and here it was complemented with CCR5 KO and peptide secretion strategy which was reported earlier and called SAVE ( Egerer et al., 2011 ). The symbol and pictogram labels (grouped for panels B, C ) are shown at the bottom of pictures.…”

“…We next hypothesized that providing the selected cells with a protective peptide would render “cured” cells resistant to repeat infection with HIV and adapted SORTS for the delivery of fusion inhibitor peptides ( Maslennikova et al., 2022 ). Three of the seven C-peptides cloned in the CD52 molecule, MT-C34, 2P23, and HP23L, displayed the strongest cell protection from HIV.…”

“…Unlike the genetic strategies intended to confer HIV resistance exclusively to edited cells, transgenic B cells protect all virus permissive cells via the secretion of bNAbs. The idea of expanding the protection against HIV-1 from edited to bystander cells via the secretion or shedding of short C-peptides ( Egerer et al., 2011 ; Maslennikova et al., 2022 ) or sCD4 ( Falkenhagen et al., 2017 ) has also been explored. However, we ( Maslennikova et al., 2022 ) and others have failed to achieve a reasonable level of protection due to problems related to the inefficient entry of a short peptide into the secretory pathway, which can partially be circumvented with peptide concatamerization ( Egerer et al., 2011 ).…”

“…The idea of expanding the protection against HIV-1 from edited to bystander cells via the secretion or shedding of short C-peptides ( Egerer et al., 2011 ; Maslennikova et al., 2022 ) or sCD4 ( Falkenhagen et al., 2017 ) has also been explored. However, we ( Maslennikova et al., 2022 ) and others have failed to achieve a reasonable level of protection due to problems related to the inefficient entry of a short peptide into the secretory pathway, which can partially be circumvented with peptide concatamerization ( Egerer et al., 2011 ). Thus, engineering B cells with CRISPR/Cas9 to produce bNAbs is a very promising technology and it would be interesting to see its effectiveness in protecting primates from HIV.…”