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a , RAG1/RAG2 expression in HIV-infected human CD4⁺ T cells following DOS treatment (single dose, 4h), showing rapid induction and complete resolution within 72 hours of treatment. MFI, mean fluorescence intensity (RAG1, n = 6 donors; RAG2, n = 5 donors). In some experiments, the T cells were pre-treated with MG-132 (1mM) prior to DOS treatment ( top right ). b , Reversible DNA-damage signalling assessed by γH2AX foci formation following rejuvenation driven HIV eradication, with resolution over time (96h) and no evidence of persistent genomic stress ( n = 6 donors). c , Stem-like T cell induction among cured T cells (CD4 + CD45RA + <t>CD27</t> + CD28 + CCR7 + , CD62L + , CD95 + ) after a one-week culture and repeated dose treatment throughout (dose escalation). Data are from n = 6 donors. d , Cell viability (Annexin V) show no increase in cell death following DOS exposure, indicating that HIV clearance is not attributable to cytotoxicity or clonal selection ( n = 3 donors). Similar RAG/ γH2AX resolution was observed following repeated (dose escalation) DOS treatment in one-week cultures. In a - d , a paired Student’s t-test. Data are shown as mean ± s.e.m. *** P < 0.001; **** P < 0.0001; ns, not significant.
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a , RAG1/RAG2 expression in HIV-infected human CD4⁺ T cells following DOS treatment (single dose, 4h), showing rapid induction and complete resolution within 72 hours of treatment. MFI, mean fluorescence intensity (RAG1, n = 6 donors; RAG2, n = 5 donors). In some experiments, the T cells were pre-treated with MG-132 (1mM) prior to DOS treatment ( top right ). b , Reversible DNA-damage signalling assessed by γH2AX foci formation following rejuvenation driven HIV eradication, with resolution over time (96h) and no evidence of persistent genomic stress ( n = 6 donors). c , Stem-like T cell induction among cured T cells (CD4 + CD45RA + <t>CD27</t> + CD28 + CCR7 + , CD62L + , CD95 + ) after a one-week culture and repeated dose treatment throughout (dose escalation). Data are from n = 6 donors. d , Cell viability (Annexin V) show no increase in cell death following DOS exposure, indicating that HIV clearance is not attributable to cytotoxicity or clonal selection ( n = 3 donors). Similar RAG/ γH2AX resolution was observed following repeated (dose escalation) DOS treatment in one-week cultures. In a - d , a paired Student’s t-test. Data are shown as mean ± s.e.m. *** P < 0.001; **** P < 0.0001; ns, not significant.
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a , RAG1/RAG2 expression in HIV-infected human CD4⁺ T cells following DOS treatment (single dose, 4h), showing rapid induction and complete resolution within 72 hours of treatment. MFI, mean fluorescence intensity (RAG1, n = 6 donors; RAG2, n = 5 donors). In some experiments, the T cells were pre-treated with MG-132 (1mM) prior to DOS treatment ( top right ). b , Reversible DNA-damage signalling assessed by γH2AX foci formation following rejuvenation driven HIV eradication, with resolution over time (96h) and no evidence of persistent genomic stress ( n = 6 donors). c , Stem-like T cell induction among cured T cells (CD4 + CD45RA + <t>CD27</t> + CD28 + CCR7 + , CD62L + , CD95 + ) after a one-week culture and repeated dose treatment throughout (dose escalation). Data are from n = 6 donors. d , Cell viability (Annexin V) show no increase in cell death following DOS exposure, indicating that HIV clearance is not attributable to cytotoxicity or clonal selection ( n = 3 donors). Similar RAG/ γH2AX resolution was observed following repeated (dose escalation) DOS treatment in one-week cultures. In a - d , a paired Student’s t-test. Data are shown as mean ± s.e.m. *** P < 0.001; **** P < 0.0001; ns, not significant.
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cd27  (Sino Biological)
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a , RAG1/RAG2 expression in HIV-infected human CD4⁺ T cells following DOS treatment (single dose, 4h), showing rapid induction and complete resolution within 72 hours of treatment. MFI, mean fluorescence intensity (RAG1, n = 6 donors; RAG2, n = 5 donors). In some experiments, the T cells were pre-treated with MG-132 (1mM) prior to DOS treatment ( top right ). b , Reversible DNA-damage signalling assessed by γH2AX foci formation following rejuvenation driven HIV eradication, with resolution over time (96h) and no evidence of persistent genomic stress ( n = 6 donors). c , Stem-like T cell induction among cured T cells (CD4 + CD45RA + <t>CD27</t> + CD28 + CCR7 + , CD62L + , CD95 + ) after a one-week culture and repeated dose treatment throughout (dose escalation). Data are from n = 6 donors. d , Cell viability (Annexin V) show no increase in cell death following DOS exposure, indicating that HIV clearance is not attributable to cytotoxicity or clonal selection ( n = 3 donors). Similar RAG/ γH2AX resolution was observed following repeated (dose escalation) DOS treatment in one-week cultures. In a - d , a paired Student’s t-test. Data are shown as mean ± s.e.m. *** P < 0.001; **** P < 0.0001; ns, not significant.
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a , RAG1/RAG2 expression in HIV-infected human CD4⁺ T cells following DOS treatment (single dose, 4h), showing rapid induction and complete resolution within 72 hours of treatment. MFI, mean fluorescence intensity (RAG1, n = 6 donors; RAG2, n = 5 donors). In some experiments, the T cells were pre-treated with MG-132 (1mM) prior to DOS treatment ( top right ). b , Reversible DNA-damage signalling assessed by γH2AX foci formation following rejuvenation driven HIV eradication, with resolution over time (96h) and no evidence of persistent genomic stress ( n = 6 donors). c , Stem-like T cell induction among cured T cells (CD4 + CD45RA + CD27 + CD28 + CCR7 + , CD62L + , CD95 + ) after a one-week culture and repeated dose treatment throughout (dose escalation). Data are from n = 6 donors. d , Cell viability (Annexin V) show no increase in cell death following DOS exposure, indicating that HIV clearance is not attributable to cytotoxicity or clonal selection ( n = 3 donors). Similar RAG/ γH2AX resolution was observed following repeated (dose escalation) DOS treatment in one-week cultures. In a - d , a paired Student’s t-test. Data are shown as mean ± s.e.m. *** P < 0.001; **** P < 0.0001; ns, not significant.

Journal: bioRxiv

Article Title: Awakening intracellular immunity for functional HIV cure

doi: 10.64898/2026.01.04.697570

Figure Lengend Snippet: a , RAG1/RAG2 expression in HIV-infected human CD4⁺ T cells following DOS treatment (single dose, 4h), showing rapid induction and complete resolution within 72 hours of treatment. MFI, mean fluorescence intensity (RAG1, n = 6 donors; RAG2, n = 5 donors). In some experiments, the T cells were pre-treated with MG-132 (1mM) prior to DOS treatment ( top right ). b , Reversible DNA-damage signalling assessed by γH2AX foci formation following rejuvenation driven HIV eradication, with resolution over time (96h) and no evidence of persistent genomic stress ( n = 6 donors). c , Stem-like T cell induction among cured T cells (CD4 + CD45RA + CD27 + CD28 + CCR7 + , CD62L + , CD95 + ) after a one-week culture and repeated dose treatment throughout (dose escalation). Data are from n = 6 donors. d , Cell viability (Annexin V) show no increase in cell death following DOS exposure, indicating that HIV clearance is not attributable to cytotoxicity or clonal selection ( n = 3 donors). Similar RAG/ γH2AX resolution was observed following repeated (dose escalation) DOS treatment in one-week cultures. In a - d , a paired Student’s t-test. Data are shown as mean ± s.e.m. *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: In some experiments, cells were evaluated for H2AX (Cell Signaling, 2595S; 1:100) and stem like reprogramming with antibodies to CD4 (PerCP5.5, BioLegend, 557956; 1:100), CCR7 (Brillant Violet 421, Biolegend, 100540; 1:100); CD27 (PerCP-Vio 700, Miltenyi; 130-120-037; 1:100); CD28 (PerCP/Cy5.5, BioLegend, 302922; 1:100); CD45RA (BV510, BioLegend, 304142; 1:100); CD62L (PE-Vio 770; Miltenyi; 130-113-621; 1:100) and CD95 (FITC, Miltenyi, 130-122-950; 1:100).

Techniques: Expressing, Infection, Fluorescence, Selection