Progranulin prevents regulatory NK cell cytotoxicity against antiviral T cells
Anfei Huang, Prashant V. Shinde, Jun Huang, Tina Senff, Haifeng C. Xu, Cassandra Margotta, Dieter Häussinger, Thomas E. Willnow, Jinping Zhang, Aleksandra A. Pandyra, Jörg Timm, Sascha Weggen, Karl S. Lang, Philipp A. Lang
Anfei Huang, Prashant V. Shinde, Jun Huang, Tina Senff, Haifeng C. Xu, Cassandra Margotta, Dieter Häussinger, Thomas E. Willnow, Jinping Zhang, Aleksandra A. Pandyra, Jörg Timm, Sascha Weggen, Karl S. Lang, Philipp A. Lang
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Research Article Immunology Infectious disease

Progranulin prevents regulatory NK cell cytotoxicity against antiviral T cells

Abstract

`NK cell–mediated regulation of antigen-specific T cells can contribute to and exacerbate chronic viral infection, but the protective mechanisms against NK cell–mediated attack on T cell immunity are poorly understood. Here, we show that progranulin (PGRN) can reduce NK cell cytotoxicity through reduction of NK cell expansion, granzyme B transcription, and NK cell–mediated lysis of target cells. Following infection with the lymphocytic choriomeningitis virus (LCMV), PGRN levels increased — a phenomenon dependent on the presence of macrophages and type I IFN signaling. Absence of PGRN in mice (Grn–/–) resulted in enhanced NK cell activity, increased NK cell–mediated killing of antiviral T cells, reduced antiviral T cell immunity, and increased viral burden, culminating in increased liver immunopathology. Depletion of NK cells restored antiviral immunity and alleviated pathology during infection in Grn–/– mice. In turn, PGRN treatment improved antiviral T cell immunity. Taken together, we identified PGRN as a critical factor capable of reducing NK cell–mediated attack of antiviral T cells.

Authors

Anfei Huang, Prashant V. Shinde, Jun Huang, Tina Senff, Haifeng C. Xu, Cassandra Margotta, Dieter Häussinger, Thomas E. Willnow, Jinping Zhang, Aleksandra A. Pandyra, Jörg Timm, Sascha Weggen, Karl S. Lang, Philipp A. Lang

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Figure 1

Progranulin limits NK cell–mediated cytotoxicity.

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Progranulin limits NK cell–mediated cytotoxicity. (A) Isolated NK cells ...
(A) Isolated NK cells were cultured with 1000 IU/ml IL-2 and indicated doses of progranulin (PGRN) for 4 days. The division of these NK cells were determined by flow cytometry (n = 3). (B) The cytotoxic activity of PGRN (100 μg/ml) treated NK cells to RMA/S cell was measured at indicated effector/target ratios (n = 4–5). (C) Granzyme B (Gzmb) and perforin (Prf1) mRNA levels from PGRN-treated (100 μg/ml) NK cells and controls were determined by qPCR (n = 6). (D) Granzyme B protein concentrations were measured in the supernatants (n = 12). (E) Mean fluorescence intensity (MFI) of granzyme B was determined in NK cells in presence of RMA/S cells with or without PGRN treatment (n = 16). (F) The expanded NK cells were treated with PGRN overnight. Gzm B MFI in NK cell was measured by flow cytometry (n = 12). (G) Splenocytes from naive WT mice were incubated with 1000 IU/ml IL-2 and/or 100 IU/ml IL-15 with or without PGRN (25 μg/ml) for 16 hours. Gzm B expression was measured by flow cytometry. The left panel represents the Gzm B+ NK cell frequency, and the right panel represents the MFI of Gzm B (n = 5). (H and I) Splenocytes from naive WT mice were incubated with 1000 U/ml IL-2, 20 ng/ml IL-12, and 5 ng/ml IL-18 with or without PGRN (25 μg/ml) for 6 hours. Gzm B (H, n = 10) and IFN-γ (I, n = 10) was determined in NK cells by flow cytometry. The upper panels represent the frequency, whereas the lower panels indicate the MFI. Data in B–I show mean ± SEM. P values calculated by 2-way ANOVA (B, C, and G–I); D, E, and F by Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.