Figure 4
Figure 4. In vivo injection of NET-loaded mDCs induces autoantibody production. Level of anti–MPO-ANCA (A), PR3-ANCA (B), ss-DNA (C), and ds-DNA (D) Ab in mice immunized with DCs loaded with NET contents in the presence or absence of DNAse, as well as in mice immunized with mDCs cocultured with apoptotic or necrotic PMNs. The induction of MPO and PR3-ANCA as well as of anti–ds-DNA autoantibodies in mice immunized with NET-loaded DCs was significantly reduced by the use of DNAse in the coculture. Immunization of mice with DCs cocultured with apoptotic PMNs induced PR3 and MPO-ANCA development but less efficiently than DCs loaded with NETotic PMNs. On the contrary, DC/apoptotic PMN immunization generated the highest titer of anti–ss-DNA Ab. Other controls were obtained by immunizing mice with DCs or PMNs alone. Each dot represents a single mouse. Median values are given. *P < .05 (1-way ANOVA with posttest Dunn correction). **P < .01 (1-way ANOVA with posttest Dunn correction). ***P < .001. Fas mutant lpr-lpr mice were used as positive control for autoantibodies.

In vivo injection of NET-loaded mDCs induces autoantibody production. Level of anti–MPO-ANCA (A), PR3-ANCA (B), ss-DNA (C), and ds-DNA (D) Ab in mice immunized with DCs loaded with NET contents in the presence or absence of DNAse, as well as in mice immunized with mDCs cocultured with apoptotic or necrotic PMNs. The induction of MPO and PR3-ANCA as well as of anti–ds-DNA autoantibodies in mice immunized with NET-loaded DCs was significantly reduced by the use of DNAse in the coculture. Immunization of mice with DCs cocultured with apoptotic PMNs induced PR3 and MPO-ANCA development but less efficiently than DCs loaded with NETotic PMNs. On the contrary, DC/apoptotic PMN immunization generated the highest titer of anti–ss-DNA Ab. Other controls were obtained by immunizing mice with DCs or PMNs alone. Each dot represents a single mouse. Median values are given. *P < .05 (1-way ANOVA with posttest Dunn correction). **P < .01 (1-way ANOVA with posttest Dunn correction). ***P < .001. Fas mutant lpr-lpr mice were used as positive control for autoantibodies.

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