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. 2003 Sep;133(3):438-47.
doi: 10.1046/j.1365-2249.2003.02228.x.

Characterization of bronchoalveolar lavage T cell subsets in sarcoidosis on the basis of CD57, CD4 and CD8

T Kurumagawa  1 , S Seki, H Kobayashi, Y Koike, S Kanoh, H Hiraide, K Motoyoshi
Affiliations

Characterization of bronchoalveolar lavage T cell subsets in sarcoidosis on the basis of CD57, CD4 and CD8

T Kurumagawa et al. Clin Exp Immunol. 2003 Sep.

Abstract

T cells expressing CD57 (a natural killer cell marker) with interferon-gamma (IFN-gamma) producing capacity increase under various conditions. CD57+ T cells are also present in the bronchoalveolar lavage fluid (BALF) of sarcoidosis, and several phenotypical and functional analyses of these cells have been reported. In the present study, BALF T cells obtained from 52 patients with sarcoidosis were classified further into CD4+CD57+ T cells, CD4+CD57- T cells, CD8+CD57+ T cells and CD8+CD57- T cells and their phenotypes and functional characteristics were assessed. Substantial proportions of these T cell subsets expressed natural killer cell markers CD161 and CD122. The biased expansion of Vbeta2 T cells was observed in both CD4+CD57+ T cells and CD4+CD57- T cells in BALF from most patients, while the expansion of other Vbeta T cells was also observed in some patients. Unexpectedly, the biased expansion of certain Vbeta T cells was also seen in either CD8+CD57+ T cells or CD8+CD57- T cells, while the expanded Vbeta T cells in CD8+ T cells differed substantially among individuals. BALF T cells showed a remarkably lower T cell receptor (TCR) intensity than that of peripheral blood T cells. Both CD8+ T cell subsets in BALF of sarcoidosis expressed the intracellular perforin/granzyme B, while all four subsets expressed intracellular IFN-gamma after in vitro activation, and CD4+ T cells, especially CD4+CD57+ T cells, expressed tumour necrosis factor-alpha. These findings indicate that CD57+ T cells as well as CD57- T cells in the BALF are phenotypically and functionally different from peripheral blood T cells and may play an important role in the Th1 dominant state and inflammation in pulmonary sarcoidosis.

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Figures

Fig. 1
Fig. 1
The surface phenotypes of T cells in BALF and PB. BALF lymphocytes and PB lymphocytes were stained with FITC-anti-CD3 antibody and with either biotin-anti-CD57 antibody developed with PC5-streptavidin, or with PE-anti-CD56 antibody (upper panels). They were also stained with FITC-anti-CD4 antibody and PC5-anti-CD8 antibody (lower panels). The numbers indicate percentage of respective populations in total lymphocytes. A representative case is shown.
Fig. 2
Fig. 2
TCR Vβ expressions in four different T cell subsets of the BALF from patients. The proportions (%) of various Vβ T cells in CD4+CD57+ T cells, CD4+CD57– T cells, CD8+CD57+ T cells and CD8+CD57– T cells were evaluated as described in the Methods section. The analyses were performed in the 20 latest consecutive patients.
Fig. 3
Fig. 3
TCR Vβ expressions in four different T cell subsets of the BALF from the controls. The proportions (%) of various Vβ T cells in CD4+CD57+ T cells, CD4+CD57– T cells, CD8+CD57+ T cells and CD8+CD57– T cells were evaluated as described in the Methods section.
Fig. 4
Fig. 4
(a) TCR intensities of BALF T cells and PB T cells. BALF lymphocytes and PB lymphocytes were stained with PC5-anti-TCRαβ antibody and with PE-anti-CD57 antibody and TCR intensities of CD57+ subsets and CD57– subsets are shown in the histograms. A representative case is shown. (b) The mean fluorescence intensities of TCRαβ were similar between the sarcoidosis patients and normal subjects irrespective of the CD57 expression.
Fig. 5.
Fig. 5.
(a) Intracellular perforin expressions of four T cell subsets in BALF. (b) Intracellular granzyme B expressions of four T cell subsets in the BALF. Intracellular perforin or granzyme B together with CD57 and CD4 or CD8 were stained in freshly isolated BALF lymphocytes as described in the Methods section and either perforin or granzyme B expression was shown in the histograms. The numbers shown in the histograms represent the percentage of positive cells. The proportions of (c) perforin- or (d) granzyme B-positive cells in four T cell subsets in the BALF. The data represent the means ± s.e.m. from eight patients. *P < 0·05, **P < 0·001.
Fig. 6
Fig. 6
Proportion of IFN-γ (a) in acute cases, (b) in chronic cases and TNF-α (c) positive cells in four T cell subsets in the BALF. BALF lymphocytes were stimulated with PMA and ionomycin, and cytokines together with CD57 and CD4 or CD8 were stained as described in the Methods section. The data represent the means ± s.e.m. from four patients of acute cases and six of chronic cases for IFN-γ and from six patients for TNF-α. *P < 0·05, **P < 0·01, ***P < 0·005, ****P < 0·0005.
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