Figure 1.

Immunopathology of herpes zoster lesions and systemic virus-specific T-cell immune responses in latently varicella-zoster virus (VZV)-infected adults. (A–D) Sections of formalin-fixed and paraffin-embedded skin biopsy obtained from a herpes zoster patient were analyzed by immunofluorescence ([IF] A, C, and D) or stained with hematoxylin and eosin (B). (A) Dual IF staining for VZV glycoprotein E ([gE] green) and CD123 (plasmacytoid dendritic cell [pDC]; red) reveals dermal pDC adjacent to VZV-infected cells. Inset: magnification of indicated area. (B) Hematoxylin and eosin staining showing polymorphonuclear cell infiltrates in a zoster vesicle. Magnification: ×100; inset: ×400. (C) Dual IF staining for VZV gE (green) and CD68 (macrophages; red) showing high numbers of dermal macrophages adjacent to VZV-infected epidermis. Insets: magnification of the indicated areas showing VZV gE-positive epidermal (pink inset) and dermal (orange inset) macrophages. (D) Dual IF staining for VZV open reading frame (ORF)63 (green) and CD3 (T cells; red) showing dermal T-cell infiltrates in proximity to VZV-infected cells. Pink inset: magnification showing VZV ORF63-positive T cell in epidermis. Orange inset: magnification showing dermal T cells adjacent to VZV ORF63-positive cells. (A, C, and D) Nuclei were counterstained with 4’,6-diamidino-2-phenylindole (blue), and dashed lines indicate epidermal—dermal junction and images are shown at ×200 magnification. (E–F) Bulk in vitro VZV-enriched CD4 (E) and CD8 (F) T-cell lines (TCLs) obtained from different representative latently VZV-infected healthy adults were assayed for reactivity to proteins encoded by individual VZV ORFs (indicated on the x-axis). (E) CD4 T-cell reactivity was determined using [³H]-thymidine incorporation, as described previously (Supplementary Data Ref. 77). Bars and error bars indicate mean ± standard deviation of duplicate measurements. Dashed horizontal line indicates threshold for positivity [(median of all negative control antigens) + (2.33 times median absolute deviation of negative control antigens)]. Both duplicates needed to exceed the threshold for the ORF to be determined as positive. (F) Interferon-γ secretion levels determined by enzyme-linked immunosorbent assay in conditioned medium of CD8 TCL cocultured with Cos-7 cells cotransfected with the responsive human leukocyte antigen (HLA)-I allele (ie, HLA-B*0802) and the individual VZV ORFs, as described previously (Supplementary Data Ref. 76). The VZV ORFs specifically recognized by T cells in the individuals analyzed are indicated.