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. 2001 Jan;75(1):544-7.
doi: 10.1128/JVI.75.1.544-547.2001.

Induction of polyomavirus-specific CD8(+) T lymphocytes by distinct dendritic cell subpopulations

D R Drake 3rd  1 , M L Shawver, A Hadley, E Butz, C Maliszewski, A E Lukacher
Affiliations

Induction of polyomavirus-specific CD8(+) T lymphocytes by distinct dendritic cell subpopulations

D R Drake 3rd et al. J Virol. 2001 Jan.

Abstract

Dendritic cells are pivotal antigen-presenting cells for generating adaptive T-cell responses. Here, we show that dendritic cells belonging to either the myeloid-related or lymphoid-related subset are permissive for infection by mouse polyomavirus and, when loaded with a peptide corresponding to the immunodominant anti-polyomavirus CD8(+) T-cell epitope or infected by polyomavirus, are each capable of driving expansion of primary polyomavirus-specific CD8(+) T-cell responses in vivo.

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Figures

FIG. 1
FIG. 1
Cell surface phenotypic analysis of LDC and MDC. (A) CD11c and CD11b expression by Nycodenz gradient-enriched, FL-expanded spleen cells. Cells in the left- and right-gated regions differentially express CD8α. (B) Plots represent cell number versus log fluorescence intensity of CD11c+ cells in the left-gated region (LDC) and in the right-gated region (MDC). Thick lines represent staining with FITC-conjugated MAbs against the indicated molecules; thin lines represent staining by FITC-conjugated isotype control MAbs.
FIG. 2
FIG. 2
MDC and LDC are permissive for polyomavirus infection. Whole cell protein lysates (50 μg/lane) from uninfected or infected (MOI of 2), FACS-sorted CD11c+ CD11b− LDC or CD11c+ CD11bhigh MDC were electrophoresed on a 10% reducing sodium dodecyl sulfate-polyacrylamide gel, transferred to nitrocellulose membranes, and immunoblotted with the anti-MT protein MAb F4 (22) (top) or rabbit anti-VP1 antiserum (33) (bottom).
FIG. 3
FIG. 3
MT389-397 peptide-pulsed or polyomavirus-infected DC of either subset induce antipolyomavirus CD8+ T cells in vivo. Uninfected, MT389-397 peptide-pulsed (10 μM), or infected LDC and MDC were injected s.c. in hind footpads. Six days later, spleen cells were stimulated directly ex vivo with MT389-397 peptide (10 μM), the non-polyomavirus Dk-binding Gag88-96 peptide (10 μM) (8), or no peptide for 6 h in the presence of brefeldin A and then stained for surface CD8 and intracellular IFN-γ. Plots are gated on CD8+ cells, and values represent the percentage of cells in the indicated region. No IFN-γ+ CD8+ cells were detected in the absence of peptide or presence of Gag88-96 peptide (data not shown). Results are representative of two experiments using two mice per group.
FIG. 4
FIG. 4
LDC and MDC migrate to draining lymph nodes after s.c. inoculation. Nycodenz gradient-enriched, FL-expanded splenic DC (2.5 × 106) were labeled with CMFDA and injected s.c. into each hind footpad of syngeneic mice; 24 h later, single-cell suspensions of popliteal lymph nodes were stained for surface CD11c and CD8α expression and analyzed by flow cytometry. Plots are gated on CMFDA+ and CMFDA− cells, and quadrants are assigned based on staining for CD11c and CD8α expression in popliteal lymph nodes from naïve mice (data not shown). Values represent the percentage of cells in the indicated region. Axes are log scale.
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