Mouse anti human cd45 percp cy5

6 week old NSG (NOD-scid IL2Rg^null) mice were obtained from The Jackson Laboratory (JAX) and at 7 weeks treated with 30mg/kg Busulfan mixed 1:1 with PBS. 24 hours later mice were injected via tail vein with 10x10⁶ human lymphocytes in 100ul 0.5% human serum albumin in PBS, comprised of 8 million CD4 T cells and 2 million CD8 T cells (NTD, BBz, 28z, or MMLV-CD4z transduced with a 50% transduction efficiency). Three weeks later mice were tail vein injected with 15ng HIV Bal mixed 1:1 with PBS. Peripheral blood was obtained by retro-orbital bleeding, and human CD4 and CAR⁺ CD8 lymphocyte counts were enumerated using BD lysis buffer and BD TruCount tubes as previously described [73 (link)], staining with mouse anti human CD45 PerCp Cy5.5 (BD Biosciences), mouse anti human CD4 BV421 (Biolegend), and mouse anti human CD8α BV711 (Biolegend).

5 week old NSG (NOD-scid IL2Rg^null) mice were obtained from The Jackson Laboratory (JAX) and at 6 weeks were injected with 5 million CD8-depleted human PBMCs, and 12 days later injected with 1 million HIV Bal-infected (or mock-infected) autologous CD4 T cells that had been in vitro infected with HIV Bal and cultured with ART for 2 days prior to freezing. The same day as HIV infection, mice began receiving 200mg/kg daily intraperitoneal injections of the reverse transcriptase inhibitor nucleotide analog tenofovir disoproxil fumarate (TDF) for 4 days. On Day 16, 5 million CD8 T cells were injected (NTD, BBz, or 28z). Peripheral blood was obtained by retro-orbital bleeding, and human CD4 and CAR⁺ CD8 lymphocyte counts were enumerated using BD lysis buffer and BD TruCount tubes as previously described [73 (link)], staining with mouse anti human CD45 PerCp Cy5.5 (BD Biosciences), mouse anti human CD4 BV421 (Biolegend), and mouse anti human CD8α BV711 (Biolegend).