Clonal expansion and TCR-independent differentiation shape the HIV-specific CD8+ effector-memory T-cell repertoire in vivo by Dirk Meyer-Olson, Brenna.

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Clonal expansion and TCR-independent differentiation shape the HIV-specific CD8+ effector-memory T-cell repertoire in vivo by Dirk Meyer-Olson, Brenna C. Simons, Joseph A. Conrad, Rita M. Smith, Louise Barnett, Shelly L. Lorey, Coley B. Duncan, Ramesh Ramalingam, and Spyros A. Kalams Blood Volume 116(3):396-405 July 22, 2010 ©2010 by American Society of Hematology

Expansions of HIV-specific TemRA phenotype cells are present to various degrees in chronic infection. Expansions of HIV-specific TemRA phenotype cells are present to various degrees in chronic infection. (A) Representative staining of CCR7 and CD45RO on CD8+ T cells in 2 representative HIV-seropositive persons. (B) Identification of the HIV-1-tetramer+ cells (red) and their corresponding Tmem phenotype (overlay: red tet+, black CD8+ T cells). (C) Comparison of effector-memory populations (paired t test) on CD8+ T cells and on HIV-1-tetramer+ T cells. Dirk Meyer-Olson et al. Blood 2010;116:396-405 ©2010 by American Society of Hematology

Analysis of effector-memory clonotype composition on HIV-specific effector-memory subsets. Analysis of effector-memory clonotype composition on HIV-specific effector-memory subsets. (A) Purification of B57-KF11 tetramer+ CD8+ T cells based on the expression of CCR7 and CD45RO in subject 10076. (B) Sequence analysis of the purified Tmem populations. (C) Analysis of the CCR7/CD45RO expression on B57-KF11 tetramer+ CD8+ T-cell expression TRBV2. Dirk Meyer-Olson et al. Blood 2010;116:396-405 ©2010 by American Society of Hematology

TCR sequences of directly sorted total tetramer+ CD8+ T cells, or tetramer+ cells sorted by effector-memory phenotype and/or CD57 expression. TCR sequences of directly sorted total tetramer+ CD8+ T cells, or tetramer+ cells sorted by effector-memory phenotype and/or CD57 expression. Dirk Meyer-Olson et al. Blood 2010;116:396-405 ©2010 by American Society of Hematology

Selective expansions of distinct HIV-epitope–specific clonotypes into terminally differentiated effector-memory phenotypes (CCR7−/CD45RA+) and association of B27-KK10 tetramer binding with expression of distinct clonotypes and effector-memory differentiation. Selective expansions of distinct HIV-epitope–specific clonotypes into terminally differentiated effector-memory phenotypes (CCR7−/CD45RA+) and association of B27-KK10 tetramer binding with expression of distinct clonotypes and effector-memory differentiation. (A) Analysis of the TRBV populations representing single clonotypes of B8-FL8 tetramer+ cells by flow cytometry. (B) TRBV usage of distinct clonotypes and CCR7/CD45RO expression on B27-KK10 tetramer cells in subject 10022. (C) Pie charts comparing the relative T-cell memory distribution of B8-FL8+ and B27-KK10+ clonotypes identified by TRBV expression in subject 10022. (D) Frequency of clonotypes within the B27-KK10 tetramer+ population in subject 10022. (E) TRBV usage and CCR7/CD45RO expression on B27-KK10 tetramer dim and bright cells in subject 10022. Dirk Meyer-Olson et al. Blood 2010;116:396-405 ©2010 by American Society of Hematology

Differential CD57 expression on HIV-1-tetramer+ CD8+ T-cell memory subsets. Differential CD57 expression on HIV-1-tetramer+ CD8+ T-cell memory subsets. (A) CD57 expression on 2 HIV-epitope specific CD8+ T-cell responses in subject 10001. (B) Lower expression of CD57 on human leukocyte antigen B8-FL8+ TemRA phenotype cells compared with TemRO phenotype cells in subject 10071. (C) Summary of CD57 expression on different memory populations on total CD8+ T cells and HIV-specific CD8+ T cells. (D) Distribution of CD57 on different memory subsets on TRBV+ and TRBV− HIV-specific CD8+ T cells in subject 10071. Dirk Meyer-Olson et al. Blood 2010;116:396-405 ©2010 by American Society of Hematology

Different expression patterns of CD57 and effector-memory marker on HIV-specific clonotypes. Different expression patterns of CD57 and effector-memory marker on HIV-specific clonotypes. Relative distribution of CD57+ (A) and TemRA-phenotype cells (B) between dominant (most frequent) clonotypes and subdominant (less frequent) clonotypes within HIV-epitope–specific CD8+ T-cell responses. (C) Relationship of CD57+ expression on the same clonotype expressing either TemRO or TemRA phenotype marker. Dirk Meyer-Olson et al. Blood 2010;116:396-405 ©2010 by American Society of Hematology