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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 <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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Human Miltenyi Biotec 130 050 101 Methoculttm Stemcell H4435 Fugene Hd Transfection Reagent Promega E2312 Kapa Rna Hyperprep Kit With Riboerase Hmr, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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