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. 2009 Aug 31;206(9):1929–1940. doi: 10.1084/jem.20090896

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© 2009 Agius et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jem.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Cellular infiltrate at the site of cutaneous DTH response is reduced in the old. Young and old volunteers were injected with 0.02 ml Candin skin antigen test and 5-mm punch biopsies were performed on days 1, 3, or 7 after injection. (A) Hematoxylin and eosin staining of skin sections from one young and one old individual on day 3 after Candin injection showing perivascular infiltrates (black arrow). The results are representative stains of three young and three old subjects that were tested. Day 3 biopsies were stained for CD3 (B) and CD4 (C) using an indirect immunoperoxidase immunohistochemical technique. The results shown are representative of five young and four old subjects and the quantification of this data are included in D. (D) The number of CD3⁺ and CD4⁺ T cells within perivascular infiltrates was determined by counting the number of positive cells within the five largest perivascular infiltrates per skin section. Mean ± SEM of 4–7 individuals per time point is shown (*, P = 0.01; **, P = 0.007; Mann-Whitney test; n = 20 old, 22 young with 4–7 volunteers per time point). (E) The percentage IFN-γ–producing antigen specific CD4⁺ T lymphocytes in old blisters is significantly reduced compared with young (n = 10 young, 5 old; **, P = 0.001, for PPD specific cells and n = 6 young, 10 old; *, P = 0.04 for C. albicans). Each symbol represents the result of a single subject. Horizontal bars represent the median. Bars, 100 µm.