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Comparative Study
. 1999 Feb;73(2):1573-9.
doi: 10.1128/JVI.73.2.1573-1579.1999.

Comparative analysis of cytotoxic T lymphocytes in lymph nodes and peripheral blood of simian immunodeficiency virus-infected rhesus monkeys

M J Kuroda  1 , J E Schmitz, W A Charini, C E Nickerson, C I Lord, M A Forman, N L Letvin
Affiliations
Comparative Study

Comparative analysis of cytotoxic T lymphocytes in lymph nodes and peripheral blood of simian immunodeficiency virus-infected rhesus monkeys

M J Kuroda et al. J Virol. 1999 Feb.

Abstract

Most studies of human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T lymphocytes (CTL) have been confined to the evaluation of these effector cells in the peripheral blood. What has not been clear is the extent to which CTL activity in the blood actually reflects this effector cell function in the lymph nodes, the major sites of HIV-1 replication. To determine the concordance between CTL activity in lymph nodes and peripheral blood lymphocytes (PBL), CTL specific for simian immunodeficiency virus of macaques (SIVmac) have been characterized in lymph nodes of infected, genetically selected rhesus monkeys by using both Gag peptide-specific functional CTL assays and tetrameric peptide-major histocompatibility complex (MHC) class I molecule complex staining techniques. In studies of six chronically SIVmac-infected rhesus monkeys, Gag epitope-specific functional lytic activity and specific tetrameric peptide-MHC class I staining were readily demonstrated in lymph node T lymphocytes. Although the numbers of tetramer-binding cells in some animals differed from those documented in their PBL, the numbers of tetramer-binding cells from these two different compartments were not statistically different. Phenotypic characterization of the tetramer-binding CD8(+) lymph node T lymphocytes of the infected monkeys demonstrated a high level of expression of the activation-associated adhesion molecules CD11a and CD49d, the Fas molecule CD95, and MHC class II-DR. These studies documented a low expression of the naive T-cell marker CD45RA and the adhesion molecule CD62L. This phenotypic profile of the tetramer-binding lymph node CD8(+) T cells was similar to that of tetramer-binding CD8(+) T cells from PBL. These observations suggest that characterization of AIDS virus-specific CTL activity by sampling of cells in the peripheral blood should provide a reasonable estimation of CTL in an individual's secondary lymphoid tissue.

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Figures

FIG. 1
FIG. 1
Tetrameric Mamu-A*01/p11C, C-M complex binds to CD8αβ+ T cells from the peripheral blood and lymph nodes of SIVmac-infected, Mamu-A*01+ rhesus monkeys. PBL and lymph node lymphocytes (LN) from six Mamu-A*01+ SIVmac-infected monkeys (GL9, 3KI, 403, 138, 575, and 154) were assessed. Flow cytometric analysis was performed with gated CD8αβ+ CD3+ T cells stained with PE-coupled tetrameric Mamu-A*01/p11C, C-M complex.
FIG. 2
FIG. 2
Tetrameric Mamu-A*01/p11C, C-M complex binds to in vitro expanded T cells from a lymph node of a SIVmac-infected, Mamu-A*01+ rhesus monkey. A whole-blood specimen (PBL) and a single-cell suspension of lymph node lymphocytes (LN) from a Mamu-A*01+ SIVmac-infected rhesus monkey (GL9) were stained with PE-coupled tetrameric Mamu-A*01/p11C, C-M complex and analyzed by flow cytometry with gating on CD8αβ+ CD3+ T cells. Cells were also expanded for 12 days in IL-2-containing medium, after stimulation either with ConA or with p11C, C-M peptide. The cells were again similarly stained and analyzed by flow cytometry.
FIG. 3
FIG. 3
Phenotypic characterization of tetrameric Mamu-A*01/p11C, C-M complex-binding CD8αβ+ T cells. A whole-blood specimen (PBL) and a single-cell suspension of lymph node lymphocytes (LN) from the Mamu-A*01+, SIVmac-infected rhesus monkey 138 were stained with Alexa 488-coupled tetrameric Mamu-A*01/p11C, C-M complex and four different PE-coupled MAbs (anti-CD28, anti-CD45RA, anti-CD62L, and anti-HLA-DR). Percentages of cells in the different quadrants are indicated.
FIG. 4
FIG. 4
Phenotypic characterization of tetrameric Mamu-A*01/p11C, C-M complex binding CD8αβ+ T cells. A whole-blood specimen (PBL) and a single-cell suspension of lymph node lymphocytes (LN) from the Mamu-A*01+, SIVmac-infected rhesus monkey 138 were stained with Alexa 488-coupled tetrameric Mamu-A*01/p11C, C-M complex and three different PE-coupled MAbs (anti-CD11a, anti-CD49d, and anti-CD95).
FIG. 5
FIG. 5
Tetrameric Mamu-A*01/p11C, C-M complex binds to CD8αβ+ and CD8αα T cells from a whole-blood specimen (PBL) and a single-cell suspension of lymph node lymphocytes (LN) of a SIVmac-infected, Mamu-A*01+ rhesus monkey. Cells from monkey 3KI were gated on CD8α+ and CD3+ T cells and were analyzed for binding of anti-CD8αβ+ and tetrameric Mamu-A*01/p11C, C-M complex.
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