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Analysis of HIV-specific TCR avidity and functional affinity . ( A ) Estimated TCR-pHLA binding strength for individual clonotypes, calculated as the average ratio of bound tetramer (antigen, Ag) UMIs to <t>CD3</t> UMIs. Statistical comparisons were performed using a non-parametric t-test. Each donor contribution is depicted with a different colour matched to the donor. ( B ). Representative FACs plot showing a monoclonal Jurkat TCR stained with a non-specific control tetramer (B*42:01-TL9) and the cognate tetramer (B*58:01-KW11). TCR-engineered T cells were co-cultured with HLA-B*58:01 monoallelic antigen-presenting cells pulsed with graded peptide concentrations (2μM to 0.125μM) at a 1:1 ratio for 18 hours. The histograms illustrate dose-dependent CD69 expression for 726_KW11-specific TCR3 clone. ( C and D ). Representative peptide dose-response curves for ( C ) IW9-specific TCRs and ( D ) KW11-specific TCRs derived from early-treated donor 726 and late-treated donor 309. TCR identifiers correspond to clonotypes with paired α and β-chains confirmed by both Illumina and Oxford nanopore sequencing. ( E and F ) Representative steady-state SPR measurements showing binding interactions between soluble HLA-B*58:01-IW9 pHLA complex and four IW9-specific TCRs, two from the early ART-treated donor 726 ( E ) and two from the late ART-treated donor 309 ( F ). Individual plots indicate donor TCR IDs and corresponding dissociation constants (K D )
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Analysis of HIV-specific TCR avidity and functional affinity . ( A ) Estimated TCR-pHLA binding strength for individual clonotypes, calculated as the average ratio of bound tetramer (antigen, Ag) UMIs to <t>CD3</t> UMIs. Statistical comparisons were performed using a non-parametric t-test. Each donor contribution is depicted with a different colour matched to the donor. ( B ). Representative FACs plot showing a monoclonal Jurkat TCR stained with a non-specific control tetramer (B*42:01-TL9) and the cognate tetramer (B*58:01-KW11). TCR-engineered T cells were co-cultured with HLA-B*58:01 monoallelic antigen-presenting cells pulsed with graded peptide concentrations (2μM to 0.125μM) at a 1:1 ratio for 18 hours. The histograms illustrate dose-dependent CD69 expression for 726_KW11-specific TCR3 clone. ( C and D ). Representative peptide dose-response curves for ( C ) IW9-specific TCRs and ( D ) KW11-specific TCRs derived from early-treated donor 726 and late-treated donor 309. TCR identifiers correspond to clonotypes with paired α and β-chains confirmed by both Illumina and Oxford nanopore sequencing. ( E and F ) Representative steady-state SPR measurements showing binding interactions between soluble HLA-B*58:01-IW9 pHLA complex and four IW9-specific TCRs, two from the early ART-treated donor 726 ( E ) and two from the late ART-treated donor 309 ( F ). Individual plots indicate donor TCR IDs and corresponding dissociation constants (K D )
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Analysis of HIV-specific TCR avidity and functional affinity . ( A ) Estimated TCR-pHLA binding strength for individual clonotypes, calculated as the average ratio of bound tetramer (antigen, Ag) UMIs to <t>CD3</t> UMIs. Statistical comparisons were performed using a non-parametric t-test. Each donor contribution is depicted with a different colour matched to the donor. ( B ). Representative FACs plot showing a monoclonal Jurkat TCR stained with a non-specific control tetramer (B*42:01-TL9) and the cognate tetramer (B*58:01-KW11). TCR-engineered T cells were co-cultured with HLA-B*58:01 monoallelic antigen-presenting cells pulsed with graded peptide concentrations (2μM to 0.125μM) at a 1:1 ratio for 18 hours. The histograms illustrate dose-dependent CD69 expression for 726_KW11-specific TCR3 clone. ( C and D ). Representative peptide dose-response curves for ( C ) IW9-specific TCRs and ( D ) KW11-specific TCRs derived from early-treated donor 726 and late-treated donor 309. TCR identifiers correspond to clonotypes with paired α and β-chains confirmed by both Illumina and Oxford nanopore sequencing. ( E and F ) Representative steady-state SPR measurements showing binding interactions between soluble HLA-B*58:01-IW9 pHLA complex and four IW9-specific TCRs, two from the early ART-treated donor 726 ( E ) and two from the late ART-treated donor 309 ( F ). Individual plots indicate donor TCR IDs and corresponding dissociation constants (K D )
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Analysis of HIV-specific TCR avidity and functional affinity . ( A ) Estimated TCR-pHLA binding strength for individual clonotypes, calculated as the average ratio of bound tetramer (antigen, Ag) UMIs to CD3 UMIs. Statistical comparisons were performed using a non-parametric t-test. Each donor contribution is depicted with a different colour matched to the donor. ( B ). Representative FACs plot showing a monoclonal Jurkat TCR stained with a non-specific control tetramer (B*42:01-TL9) and the cognate tetramer (B*58:01-KW11). TCR-engineered T cells were co-cultured with HLA-B*58:01 monoallelic antigen-presenting cells pulsed with graded peptide concentrations (2μM to 0.125μM) at a 1:1 ratio for 18 hours. The histograms illustrate dose-dependent CD69 expression for 726_KW11-specific TCR3 clone. ( C and D ). Representative peptide dose-response curves for ( C ) IW9-specific TCRs and ( D ) KW11-specific TCRs derived from early-treated donor 726 and late-treated donor 309. TCR identifiers correspond to clonotypes with paired α and β-chains confirmed by both Illumina and Oxford nanopore sequencing. ( E and F ) Representative steady-state SPR measurements showing binding interactions between soluble HLA-B*58:01-IW9 pHLA complex and four IW9-specific TCRs, two from the early ART-treated donor 726 ( E ) and two from the late ART-treated donor 309 ( F ). Individual plots indicate donor TCR IDs and corresponding dissociation constants (K D )

Journal: bioRxiv

Article Title: Duration of Initial Viremia Modulates Functional Properties of HIV-specific T Cell Receptors

doi: 10.64898/2026.01.29.702605

Figure Lengend Snippet: Analysis of HIV-specific TCR avidity and functional affinity . ( A ) Estimated TCR-pHLA binding strength for individual clonotypes, calculated as the average ratio of bound tetramer (antigen, Ag) UMIs to CD3 UMIs. Statistical comparisons were performed using a non-parametric t-test. Each donor contribution is depicted with a different colour matched to the donor. ( B ). Representative FACs plot showing a monoclonal Jurkat TCR stained with a non-specific control tetramer (B*42:01-TL9) and the cognate tetramer (B*58:01-KW11). TCR-engineered T cells were co-cultured with HLA-B*58:01 monoallelic antigen-presenting cells pulsed with graded peptide concentrations (2μM to 0.125μM) at a 1:1 ratio for 18 hours. The histograms illustrate dose-dependent CD69 expression for 726_KW11-specific TCR3 clone. ( C and D ). Representative peptide dose-response curves for ( C ) IW9-specific TCRs and ( D ) KW11-specific TCRs derived from early-treated donor 726 and late-treated donor 309. TCR identifiers correspond to clonotypes with paired α and β-chains confirmed by both Illumina and Oxford nanopore sequencing. ( E and F ) Representative steady-state SPR measurements showing binding interactions between soluble HLA-B*58:01-IW9 pHLA complex and four IW9-specific TCRs, two from the early ART-treated donor 726 ( E ) and two from the late ART-treated donor 309 ( F ). Individual plots indicate donor TCR IDs and corresponding dissociation constants (K D )

Article Snippet: The sorted mCherry-positive cells were cultured in R10 media for 7 days, after which CD3 + cells were isolated using the REAlease CD3 Microbead Kit, (Miltenyl Biotec).

Techniques: Functional Assay, Binding Assay, Staining, Control, Cell Culture, Expressing, Derivative Assay, Nanopore Sequencing

( A ) IW9 and ( B ) KW11-specific TCRs. The EC 50 for CD69 expression profile from peptide dose-response assays for each clonotype is on the y axis, and the corresponding tetramer/CD3 UMI ratio on the x-axis. Red dots indicate early ART-treated donors, and blue dots indicate late ART-treated donors.

Journal: bioRxiv

Article Title: Duration of Initial Viremia Modulates Functional Properties of HIV-specific T Cell Receptors

doi: 10.64898/2026.01.29.702605

Figure Lengend Snippet: ( A ) IW9 and ( B ) KW11-specific TCRs. The EC 50 for CD69 expression profile from peptide dose-response assays for each clonotype is on the y axis, and the corresponding tetramer/CD3 UMI ratio on the x-axis. Red dots indicate early ART-treated donors, and blue dots indicate late ART-treated donors.

Article Snippet: The sorted mCherry-positive cells were cultured in R10 media for 7 days, after which CD3 + cells were isolated using the REAlease CD3 Microbead Kit, (Miltenyl Biotec).

Techniques: Expressing