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Flexibility of the HIV-specific T-cell receptor repertoire is a hallmark of HIV-1 infection. Altered differentiation of HIV-specific CD45RO⁺/CCR7⁻ (TemRO) CD8⁺ effector-memory T cells into CD45RA⁺/CCR7⁻ (TemRA) CD8⁺ effector-memory T cells as well as increased expression of the senescence marker CD57 has been frequently observed HIV-1 infection, but the structural relationship between clonal expansion and T-cell differentiation has not been defined. In this study, we demonstrate that HIV-specific clonotypes have differing degrees of TemRA differentiation but always maintain a significant proportion of TemRO-phenotype cells. These data indicate that structural constraints of the TCR/peptide major histocompatibility complex interaction play a central role in the TemRA differentiation of HIV-specific CD8⁺ T cells in chronic HIV-1 infection. Clonotypes with a predominantly TemRA phenotype had a …