Distinct activation of primary human BDCA1+ dendritic cells upon interaction with stressed or infected β cells
B. M. Schulte
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands
Search for more papers by this authorE. D. Kers-Rebel
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands
Search for more papers by this authorR. Bottino
Department of Pediatrics, Diabetes Institute, University of Pittsburgh, Pittsburgh, PA, USA
Search for more papers by this authorJ. D. Piganelli
Department of Pediatrics, Diabetes Institute, University of Pittsburgh, Pittsburgh, PA, USA
Search for more papers by this authorJ. M. D. Galama
Department of Medical Microbiology, Radboud University Medical Center, Nijmegen
Search for more papers by this authorM. A. Engelse
Department of Nephrology, Leiden University Medical Center, Leiden
Search for more papers by this authorE. J. P. de Koning
Department of Nephrology, Leiden University Medical Center, Leiden
Department of Endocrinology, Leiden University Medical Center, Leiden
Hubrecht Institute, Utrecht, the Netherlands
Search for more papers by this authorCorresponding Author
G. J. Adema
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands
Correspondence: G. J. Adema, Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA, Nijmegen, the Netherlands. E-mail: [email protected]Search for more papers by this authorB. M. Schulte
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands
Search for more papers by this authorE. D. Kers-Rebel
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands
Search for more papers by this authorR. Bottino
Department of Pediatrics, Diabetes Institute, University of Pittsburgh, Pittsburgh, PA, USA
Search for more papers by this authorJ. D. Piganelli
Department of Pediatrics, Diabetes Institute, University of Pittsburgh, Pittsburgh, PA, USA
Search for more papers by this authorJ. M. D. Galama
Department of Medical Microbiology, Radboud University Medical Center, Nijmegen
Search for more papers by this authorM. A. Engelse
Department of Nephrology, Leiden University Medical Center, Leiden
Search for more papers by this authorE. J. P. de Koning
Department of Nephrology, Leiden University Medical Center, Leiden
Department of Endocrinology, Leiden University Medical Center, Leiden
Hubrecht Institute, Utrecht, the Netherlands
Search for more papers by this authorCorresponding Author
G. J. Adema
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands
Correspondence: G. J. Adema, Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA, Nijmegen, the Netherlands. E-mail: [email protected]Search for more papers by this authorSummary
Derailment of immune responses can lead to autoimmune type 1 diabetes, and this can be accelerated or even induced by local stress caused by inflammation or infection. Dendritic cells (DCs) shape both innate and adaptive immune responses. Here, we report on the responses of naturally occurring human myeloid BDCA1+ DCs towards differentially stressed pancreatic β cells. Our data show that BDCA1+ DCs in human pancreas-draining lymph node (pdLN) suspensions and blood-derived BDCA1+ DCs both effectively engulf β cells, thus mimicking physiological conditions. Upon uptake of enterovirus-infected, but not mock-infected cells, BDCA1+ DCs induced interferon (IFN)-α/β responses, co-stimulatory molecules and proinflammatory cytokines and chemokines. Notably, induction of stress in β cells by ultraviolet irradiation, culture in serum-free medium or cytokine-induced stress did not provoke strong DC activation, despite efficient phagocytosis. DC activation correlated with the amount of virus used to infect β cells and required RNA within virally infected cells. DCs encountering enterovirus-infected β cells, but not those incubated with mock-infected or stressed β cells, suppressed T helper type 2 (Th2) cytokines and variably induced IFN-γ in allogeneic mixed lymphocyte reaction (MLR). Thus, stressed β cells have little effect on human BDCA1+ DC activation and function, while enterovirus-infected β cells impact these cells significantly, which could help to explain their role in development of autoimmune diabetes in individuals at risk.
Supporting Information
Additional Supporting information may be found in the online version of this article at the publisher's web-site:
| Filename | Description |
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| cei12779-sup-0001-suppinfo1.ppt2.4 MB |
Fig. S1. Efficient uptake of mock-, virus-infected and stressed β cells by blood-derived cell antigen 1 (BDCA1+) dendritic cells (DCs). (a) Min6 cells were mock- or Coxsackie B virus (CVB)-infected or exposed to indicated stress and Min6 viability was determined prior to co-culture with BDCA1+ DCs using viability dye. (b) Blood-derived BDCA1+ DCs were co-cultured with PKH67-labelled mock- or CVB-infected Min6 cells, stressed Min6 cells, were stimulated with poly I:C or left unstimulated, and uptake was determined as in Fig. 1c. Fig. S2. Blood-derived cell antigen 1 (BDCA1+) dendritic cells (DCs) that encounter Coxsackie B virus (CVB)-infected, but not mock-infected Min6 cells induce type I interferon (IFN) responses. (a) DCs were co-cultured with mock- or CVB-infected Min6 cells, stimulated with poly I:C or left unstimulated (medium) and after overnight culture supernatant was harvested and assessed for IFN-α. (b) DCs were stimulated as in (a) or exposed to CVB3 and after 6 h RNA was harvested and mRNA expression was analysed. (c) DCs were stimulated as in (b) and protein expression was analysed after overnight culture. Average ± standard error of the mean (s.e.m.) three (a), seven (b) experiments or representative of three experiments (d). * P < 0·5; **P < 0·01; ***P < 0·001 as determined by one-way analysis of variance (anova) and post-hoc Tukey analysis. Fig. S3. Phenotypical dendritic cell (DC) maturation and production of proinflammatory cytokines and chemokines by blood-derived cell antigen 1 (BDCA1+) DCs that engulf Coxsackie B virus (CVB)-infected, but not mock-infected Min6 cells. (a) DCs cultured as in Fig. S2b were analysed for indicated cell surface markers after overnight culture. (b,c) Supernatant of cells cultured in (a) is analysed for indicated cytokines and chemokines. Whisker plot for more than 16 experiments (a) or column scatterplot from nine different donors (b,c). Corresponding symbols represent the same donor in within a figure (b,c). *P < 0·5; **P < 0·01; ***P < 0·001 as determined by one-way analysis of variance (anova) and post-hoc Tukey analysis. Fig. S4. Blood-derived cell antigen 1 (BDCA1+) dendritic cells (DCs) stimulated with Coxsackie B virus (CVB)-infected, but not mock-infected Min6 cells, induce T cells with T helper type 1 (Th1) phenotype while suppressing Th2 responses. Supernatant from mixed lymphocyte reaction (MLR) cultures using indicated stimuli was analysed for cytokine production 48 h after start of MLR. Shown is average ± standard error of the mean (s.e.m.) of five different donors. **P < 0·01 as determined by one-way analysis of variance (anova) and post-hoc Tukey analysis. Fig. S5. Induction of interferon (IFN)-stimulated genes in Coxsackie B virus (CVB)-infected human islets of Langerhans. Human islets of Langerhans were mock- or CVB-infected and protein expression was analysed after 48 h. hIsl/M = mock-infected human islets of Langerhans; hIsl/CVB = CVB-infected human islets of Langerhans. Fig. S6. Cytokine and chemokine production within one blood-derived cell antigen 1 (BDCA1+) dendritic cell (DC) donor upon co-culture with Min6 cells or frozen and thawed lysate of islets of Langerhans. DCs from one donor were cultured as in Fig. 3a or co-cultured with frozen and thawed lysate of mock- or Coxsackie B virus (CVB)-infected human islets of Langerhans. Cytokines and chemokines were analysed as for Fig. 3b,c. Fig. S7. Cytokine and chemokine production upon co-culture of blood-derived cell antigen 1 (BDCA1+) dendritic cells (DCs) with frozen and thawed lysates of mock- or Coxsackie B virus (CVB)-infected human islets of Langerhans. DCs were cultured and analysed as in Fig. S6. |
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