HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death
Scott J. Brodie, Bruce K. Patterson, Deborah A. Lewinsohn, Kurt Diem, David Spach, Phillip D. Greenberg, Stanley R. Riddell, Lawrence Corey
Scott J. Brodie, Bruce K. Patterson, Deborah A. Lewinsohn, Kurt Diem, David Spach, Phillip D. Greenberg, Stanley R. Riddell, Lawrence Corey
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Article

HIV-specific cytotoxic T lymphocytes traffic to lymph nodes and localize at sites of HIV replication and cell death

Abstract

We have tracked the in vivo migration and have identified in vivo correlates of cytotoxic T-lymphocyte (CTL) activity in HIV-seropositive subjects infused with autologous gene-marked CD8+ HIV-specific CTL. The number of circulating gene-marked CTL ranged from 1.6 to 3.5% shortly after infusion to less than 0.5% 2 weeks later. Gene-marked CTL were present in the lymph node at 4.5- to 11-fold excess and colocalized within parafollicular regions of the lymph node adjacent to cells expressing HIV tat fusion transcripts, a correlate of virus replication. The CTL clones expressed the CCR5 receptor and localized among HIV-infected cells expressing the ligands MIP-1α and MIP-1β, CC-chemokines produced at sites of virus replication. Aggregates of apoptotic cells and cells expressing granzyme-B localized within these same sites. In contrast, lymph node sections from untreated HIV-seropositive subjects, all with significant viral burden (> 50,000 HIV RNA copies/mL plasma), showed no CC-chemokine expression and exhibited only sporadic and randomly distributed cells expressing granzymes and/or apoptotic cells. These studies show that the infused CTL specifically migrate to sites of HIV replication and retain their antigen-specific cytolytic potential. Moreover, these studies provide a methodology that will facilitate studies of both the magnitude and functional phenotype of Ag-specific CD8+ T cells in vivo.

Authors

Scott J. Brodie, Bruce K. Patterson, Deborah A. Lewinsohn, Kurt Diem, David Spach, Phillip D. Greenberg, Stanley R. Riddell, Lawrence Corey

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Figure 6

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Representative histologic sections of inguinal lymph node collected from...
Representative histologic sections of inguinal lymph node collected from patient 2 by excisional biopsy 4 days after the second infusion of neo-modified HIV-specific CTL. Serial sections were analyzed for sites of localization of neo-modified CD8+ CTL (PCR-ISH) (a), cells expressing granzyme-B (anti-GrB mAb) (b), cells expressing HIV tat fusion transcripts (RT-PCR-ISH) (c), cells expressing the CC-chemokine MIP-1β (anti-human MIP-1β mAb) (d), and cells showing DNA fragmentation (TUNEL “apoptosis” assay) (e). Dual stains were used to identify the proportion of CD4+ (anti-CD4 mAb + TUNEL) (f; inset shows higher magnification of dual-stained cells [arrows]) and CD8+ (anti-CD8 mAb + TUNEL) (g) T lymphocytes demonstrating apoptosis. The HIV-specific CTL expressed the cell surface receptors CD8 and CCR5 and could be stimulated to express granzyme-B before their infusion (h: anti-CD8 mAb (left), anti-CCR5 mAb (middle), anti-GrB mAb (right). Antibody labels were detected by immunoperoxidase staining with DAB as the chromogen and apoptotic cells detected by TUNEL, followed by anti-DIG mAb, and developed with BCIP/NBT. f, follicular germinal centers. Bar, 100 μm.