mouse lung tissue il 11 levels  (R&D Systems)

Journal: iScience

Article Title: Discovery of a novel SHIP1 agonist that promotes degradation of lipid-laden phagocytic cargo by microglia

doi: 10.1016/j.isci.2022.104170

Figure Lengend Snippet: K306 has biological activity consistent with SHIP1 agonistic activity and is a more potent agonist than the pelorol AQX-MN100 (A) IL-6 and (B)TNF-α production by BV2 microglia cells stimulated with LPS for 6 h or 2 h, respectively, as measured from supernatants by ELISA. (Representative results of two independent comparisons of all potential agonists at 5μM). All cells were treated with agonists or vehicle control 1 h before LPS challenge. Bars indicate mean ±SEM. Statistical analysis was performed with one-way ANOVA with Dunnett correction for multiple comparisons versus control (DMSO), ∗∗p < 0.01, ∗p < 0.05). 5′ fluorescence polarization Assay (FP) to measure PtdIns(3,4,5)P 3 5′ phosphatase activity on (C) tSHIP1 with K306 and MN-100 and (D) SHIP1-Enzyme (S1-Enz) and SHIP2-Enzyme (S2-Enz) with K306. EC 50 of K306 and MN-100 on tSHIP1 and the EC 50 of K306 on S1-Enz and S2-Enz were calculated by incubating dilutions of K306 or MN-100 with either enzyme for 20 min at 37°C. PI(3,4)P 2 generated by the SHIP enzymes is then measured using the FP assay (Echelon Biosciences). Control reactions include probe alone (PA), where all the probe is free rotating and thus not polarized and no-enzyme (NE) control, where the probe is not displaced from the detector by enzymatically generated PI(3,4)P 2 and highly polarized. Shown is one of 2 independent experiments with 6 replicate wells/compound concentration. EC 50 was calculated with Prizm version 9.3.1, using Nonlinear regression [Agonist] vs. response (three parameters), using Robust Fitting method, Medium Convergence Criteria, No Weighting and Considering every Y replicate as an individual point.

Article Snippet: TNF-α and IL-6 levels were normalized to percentage of live cells as compared to control (vehicle) determined by the Cell Counting KIT-8 (CCK-8, Dojindo Molecular Technologies Inc), that was diluted 1:10 in media, incubated with the cells for 2h and detected at 460nm with a plate reader (Synergy 2, BioTek). following manufacturer’s recommendations.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Fluorescence, Generated, FP Assay, Concentration Assay

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: (A) Published data by our group showing the total number of larvae found in the lungs in different days of infection summarizing the kinetics of Ascaris larval migration in the experimentally infected mice lungs . (B) Transcriptional profile of Ascaris -infected lungs of mice at day 8 post-infection (peak of larval burden in the lungs). Volcano plot shows the significant upregulated and downregulated genes, with the log2 fold change, in the lungs of Ascaris -infected mice over naïve lungs obtained from a nanostring analysis of 507 genes associated with inflammation. Dotted line indicates the p-value threshold in 0.05. Genes labelled in red are differentially expressed genes (DEG) associated with eosinophil activation pathway. Genes labelled in green are DEG associated with neutrophil activation pathway. Line graph showing the profile of cytokine/chemokine levels (mean ± SEM) in the lung tissue homogenate of C57/BL-6 mice during different time points of pulmonary larval migration, including IL-6 (C), IL-11 (D), G-CSF (E), CXCL-1 (F). Dotted line in each graph represents the baseline levels of each analyte in naïve lung homogenates. P values are represented in the graph in comparison to naïve lungs. *p < 0.05, **p < 0.01. *p < 0.001. Scatter plots showing the parasite burden in the lungs of C57BL6 mice infected with different loads of Ascaris eggs (G) and the respective IL-11 protein levels in the lungs of these mice at day 8p.i (H), followed by the Spearman correlation analysis between number of larvae trafficking in the lungs and the levels of IL-11 (pg/mL) (I). p and r values are indicated in the graphs. Differences between naïve and Ascaris -infected groups were considered statistically significant when P < 0.05. Data are representative of 2 independent experiments with n = 5 for each group per experiment.

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: Infection, Migration, Activation Assay, Comparison

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Immunophenotypic analysis of CD45 + lung myeloid cells on the course of larval migration in the lungs of Ascaris -infected (n = 5) and naïve (n = 5) C57/BL6 mice. Representative flow cytometry dot plots showing the frequency of CD11b + Ly6G + neutrophils at day 8p.i (A), SiglecF + CD11ceosinophils at day 18p.i (B) and CD11b + F4/80 + macrophages at day 18p.i (B) in the lungs, and their associate scatter dot plots with their absolute numbers at day 0, 5, 8 and 18p.i. (D) Flow cytometry dot plot of Ascaris -infected lung cells showing the frequency of Lineage − CD11b + CD24 + neutrophils at day 8p.i of C57/BL6 mice treated with isotype control (n = 5) or anti-Ly6G antibodies (n = 5). IL-6 and IL-11 levels at day 8p.i in the neutrophil-depleted Ascaris -infected lung homogenates in comparison with the isotype control. Data is represented by the geometric mean, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05. Data are representative of 2 independent experiments with n = 5 for each group per experiment.

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: Migration, Infection, Flow Cytometry, Control, Comparison

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: (A) Using publicly available single-nucleus RNA-sequencing data from naïve lung cells , a total of 31 clusters of cells in the lung tissue of C57BL6 mice were identified based on their expression profile using Seurat and annotated fine cell types on the ImmGen mouse database using SingleR. (B) Expression level of IL-11 was investigated among the clusters. The frequency of IL-11 + lung cells deconvoluted by cell type among the four major subsets including epithelial, stromal, endothelial and immune cells (C). Confocal imaging analysis of naïve (panels a and b) and Ascaris -infected lungs (c and d) staining with anti-CD45 cells for immune cells (green), anti-EpCAM for epithelial cells (red), intracellular anti-IL-11 for IL-11 expression (yellow) and DAPI for nuclei (blue).

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: RNA Sequencing, Expressing, Imaging, Infection, Staining

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Fresh-frozen lung tissue section of four naïve and four Ascaris -infected WT mice were hybridized with probes to Il11 and probes to either Epcam or Pecam (probe set 1) (A), or probes to Acta2 and Pdgfra (probe set 3 (B)]. Nuclei were counterstained with DAPI (blue). Scale bar = 20 μm. The frequency of IL-11 expressing Epcam + epithelial cells (C), Pecam + endothelial cells (D), Pdgfrα + fibroblasts (E), Acta2 + smooth muscle cells (F) and Acta2 + Pdgfrα + cells (myofibroblasts) (G) were calculated based in the total number of IL11 + DAPI + cells. Data is represented by the geometric mean, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05.

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: Infection, Expressing

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Recombinant murine IL-11 (2 μg/day) or PBS were daily administered intranasally for 7 days in both naïve (n = 5) and Ascaris -infected (n = 5) C57/BL-6 mice. Parasite burden and cytokine/chemokine profiling were performed in the lungs at day 8p.i (A). IL-11 levels were measured in the lungs homogenates of all four groups (B). Heatmap analysis showing the transformed z-score data of cytokines (C), growth-factors (D) and chemokines (E) levels (pg/mL) in the lung’s homogenate of both naïve and Ascaris -infected C57/BL-6 mice that received either PBS or rIL-11. Scatter plots showing IL-6, CXCL-1, G-CSF and CXCL-5 in the lung homogenate of naïve mice (F) and Ascaris -infected mice (G) that received either PBS or rIL-11. (H) Representative histological analysis of lung section stained with H&E from Ascaris -infected mice at day 8p.i treated daily with PBS (a-c) or treated daily with rIL-11 (d-f). Parameters such as peribronchial (*) and perivascular cellular infiltration (#) were used to calculate the total pulmonary inflammation score (I). Septum thickness (arrows), Ascaris larva (arrowhead) and hemorrhagic areas (h) are also indicated in the panels a and d. Frequency of Ly6G + neutrophils by flow cytometry in all four groups of analysis (J). Data is represented by the geometric mean, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05. Data are representative of 2 independent experiments with n = 5 for each group per experiment.

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: Recombinant, Infection, Transformation Assay, Staining, Flow Cytometry

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Both mouse bronchial EpCAM + epithelial cells (Cell Biologics, Inc) and mouse bronchial fibroblasts MM14.Lu (ATCC) were cultured at 1 × 10 5 cells/well and stimulated in the absence (media) or presence of 100 ng/mL of recombinant murine cytokines for 24 h at 37 °C in 5 % CO 2 . After incubation, IL-11 levels were measured in the culture supernatant (A). Both cell types were also stimulated in absence or in the presence of recombinant mouse TGF-b1 at different concentrations (1, 10, 100, 1000 ng/mL) in the same culture conditions as above, for IL-11 quantification in the culture supernatant (B). Finally, both mouse bronchial fibroblasts and epithelial cells were stimulated in the same conditions with recombinant mouse IL-11 in a dose–response manner (0, 0.2, 2, 20 and 200 ng/mL) for the quantification of CXCL-1 (C), IL-6 (D) and G-CSF (E) levels in the culture supernatant. Scatter plots represent the geometric mean of four replicates, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05. Data are representative of 2 independent in vitro experiments with 3–4 replicates each.

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: Cell Culture, Recombinant, Incubation, In Vitro

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Ascaris larval migration from the pulmonary circulation into the lung parenchyma and airways induces mechanical and inflammatory injury to endothelial and epithelial barriers. This tissue damage triggers the rapid upregulation of IL-11 within the lung microenvironment. Based on our data, EpCAM + epithelial cells, PECAM + endothelial cells, and peribronchial stromal populations, including fibroblasts, myofibroblasts, pericytes and airway smooth muscle cells, represent the major sources of IL-11 in helminth-infected lungs. Fibroblasts, and likely differentiated myofibroblasts, respond to TGF-β signaling by inducing IL-11 expression, consistent with established profibrotic pathways. In contrast, the mechanisms by which larval stages directly stimulate epithelial, endothelial, and smooth muscle cells remain unclear. Here, we hypothesize that these structural cell populations act as early responders to tissue damage, rapidly upregulating IL-11 and positioning it as an alarmin-like mediator that amplifies downstream inflammatory responses during acute infection. IL-11 signaling within epithelial and stromal compartments promotes the production of neutrophil-associated mediators, including CXCL-1, G-CSF, and IL-6, leading to an early and robust recruitment of neutrophils from the circulation into lung tissue. Recruited neutrophils, themselves a major source of IL-6, further contribute to sustained pulmonary inflammation. Notably, IL-11-producing cell populations express high levels of IL-11Rα1, supporting a model in which IL-11 signaling is reinforced through autocrine and paracrine feedback loops within damaged lung niches during helminth infection. Figure created with BioRender.

Article Snippet: The mouse lung tissue IL-11 levels were measured by using a Duoset ELISA Kit (R&D Systems, Minneapolis, Minn).

Techniques: Migration, Infection, Expressing

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: (A) Published data by our group showing the total number of larvae found in the lungs in different days of infection summarizing the kinetics of Ascaris larval migration in the experimentally infected mice lungs . (B) Transcriptional profile of Ascaris -infected lungs of mice at day 8 post-infection (peak of larval burden in the lungs). Volcano plot shows the significant upregulated and downregulated genes, with the log2 fold change, in the lungs of Ascaris -infected mice over naïve lungs obtained from a nanostring analysis of 507 genes associated with inflammation. Dotted line indicates the p-value threshold in 0.05. Genes labelled in red are differentially expressed genes (DEG) associated with eosinophil activation pathway. Genes labelled in green are DEG associated with neutrophil activation pathway. Line graph showing the profile of cytokine/chemokine levels (mean ± SEM) in the lung tissue homogenate of C57/BL-6 mice during different time points of pulmonary larval migration, including IL-6 (C), IL-11 (D), G-CSF (E), CXCL-1 (F). Dotted line in each graph represents the baseline levels of each analyte in naïve lung homogenates. P values are represented in the graph in comparison to naïve lungs. *p < 0.05, **p < 0.01. *p < 0.001. Scatter plots showing the parasite burden in the lungs of C57BL6 mice infected with different loads of Ascaris eggs (G) and the respective IL-11 protein levels in the lungs of these mice at day 8p.i (H), followed by the Spearman correlation analysis between number of larvae trafficking in the lungs and the levels of IL-11 (pg/mL) (I). p and r values are indicated in the graphs. Differences between naïve and Ascaris -infected groups were considered statistically significant when P < 0.05. Data are representative of 2 independent experiments with n = 5 for each group per experiment.

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: Infection, Migration, Activation Assay, Comparison

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Immunophenotypic analysis of CD45 + lung myeloid cells on the course of larval migration in the lungs of Ascaris -infected (n = 5) and naïve (n = 5) C57/BL6 mice. Representative flow cytometry dot plots showing the frequency of CD11b + Ly6G + neutrophils at day 8p.i (A), SiglecF + CD11ceosinophils at day 18p.i (B) and CD11b + F4/80 + macrophages at day 18p.i (B) in the lungs, and their associate scatter dot plots with their absolute numbers at day 0, 5, 8 and 18p.i. (D) Flow cytometry dot plot of Ascaris -infected lung cells showing the frequency of Lineage − CD11b + CD24 + neutrophils at day 8p.i of C57/BL6 mice treated with isotype control (n = 5) or anti-Ly6G antibodies (n = 5). IL-6 and IL-11 levels at day 8p.i in the neutrophil-depleted Ascaris -infected lung homogenates in comparison with the isotype control. Data is represented by the geometric mean, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05. Data are representative of 2 independent experiments with n = 5 for each group per experiment.

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: Migration, Infection, Flow Cytometry, Control, Comparison

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: (A) Using publicly available single-nucleus RNA-sequencing data from naïve lung cells , a total of 31 clusters of cells in the lung tissue of C57BL6 mice were identified based on their expression profile using Seurat and annotated fine cell types on the ImmGen mouse database using SingleR. (B) Expression level of IL-11 was investigated among the clusters. The frequency of IL-11 + lung cells deconvoluted by cell type among the four major subsets including epithelial, stromal, endothelial and immune cells (C). Confocal imaging analysis of naïve (panels a and b) and Ascaris -infected lungs (c and d) staining with anti-CD45 cells for immune cells (green), anti-EpCAM for epithelial cells (red), intracellular anti-IL-11 for IL-11 expression (yellow) and DAPI for nuclei (blue).

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: RNA Sequencing, Expressing, Imaging, Infection, Staining

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Fresh-frozen lung tissue section of four naïve and four Ascaris -infected WT mice were hybridized with probes to Il11 and probes to either Epcam or Pecam (probe set 1) (A), or probes to Acta2 and Pdgfra (probe set 3 (B)]. Nuclei were counterstained with DAPI (blue). Scale bar = 20 μm. The frequency of IL-11 expressing Epcam + epithelial cells (C), Pecam + endothelial cells (D), Pdgfrα + fibroblasts (E), Acta2 + smooth muscle cells (F) and Acta2 + Pdgfrα + cells (myofibroblasts) (G) were calculated based in the total number of IL11 + DAPI + cells. Data is represented by the geometric mean, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05.

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: Infection, Expressing

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Recombinant murine IL-11 (2 μg/day) or PBS were daily administered intranasally for 7 days in both naïve (n = 5) and Ascaris -infected (n = 5) C57/BL-6 mice. Parasite burden and cytokine/chemokine profiling were performed in the lungs at day 8p.i (A). IL-11 levels were measured in the lungs homogenates of all four groups (B). Heatmap analysis showing the transformed z-score data of cytokines (C), growth-factors (D) and chemokines (E) levels (pg/mL) in the lung’s homogenate of both naïve and Ascaris -infected C57/BL-6 mice that received either PBS or rIL-11. Scatter plots showing IL-6, CXCL-1, G-CSF and CXCL-5 in the lung homogenate of naïve mice (F) and Ascaris -infected mice (G) that received either PBS or rIL-11. (H) Representative histological analysis of lung section stained with H&E from Ascaris -infected mice at day 8p.i treated daily with PBS (a-c) or treated daily with rIL-11 (d-f). Parameters such as peribronchial (*) and perivascular cellular infiltration (#) were used to calculate the total pulmonary inflammation score (I). Septum thickness (arrows), Ascaris larva (arrowhead) and hemorrhagic areas (h) are also indicated in the panels a and d. Frequency of Ly6G + neutrophils by flow cytometry in all four groups of analysis (J). Data is represented by the geometric mean, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05. Data are representative of 2 independent experiments with n = 5 for each group per experiment.

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: Recombinant, Infection, Transformation Assay, Staining, Flow Cytometry

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Both mouse bronchial EpCAM + epithelial cells (Cell Biologics, Inc) and mouse bronchial fibroblasts MM14.Lu (ATCC) were cultured at 1 × 10 5 cells/well and stimulated in the absence (media) or presence of 100 ng/mL of recombinant murine cytokines for 24 h at 37 °C in 5 % CO 2 . After incubation, IL-11 levels were measured in the culture supernatant (A). Both cell types were also stimulated in absence or in the presence of recombinant mouse TGF-b1 at different concentrations (1, 10, 100, 1000 ng/mL) in the same culture conditions as above, for IL-11 quantification in the culture supernatant (B). Finally, both mouse bronchial fibroblasts and epithelial cells were stimulated in the same conditions with recombinant mouse IL-11 in a dose–response manner (0, 0.2, 2, 20 and 200 ng/mL) for the quantification of CXCL-1 (C), IL-6 (D) and G-CSF (E) levels in the culture supernatant. Scatter plots represent the geometric mean of four replicates, and the p-values are indicated in each graph. Differences between the groups were considered statistically significant when P < 0.05. Data are representative of 2 independent in vitro experiments with 3–4 replicates each.

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: Cell Culture, Recombinant, Incubation, In Vitro

Journal: Mucosal immunology

Article Title: IL-11 acts as an alarmin-like pro-inflammatory mediator regulating mucosal responses during helminth infection

doi: 10.1016/j.mucimm.2026.01.005

Figure Lengend Snippet: Ascaris larval migration from the pulmonary circulation into the lung parenchyma and airways induces mechanical and inflammatory injury to endothelial and epithelial barriers. This tissue damage triggers the rapid upregulation of IL-11 within the lung microenvironment. Based on our data, EpCAM + epithelial cells, PECAM + endothelial cells, and peribronchial stromal populations, including fibroblasts, myofibroblasts, pericytes and airway smooth muscle cells, represent the major sources of IL-11 in helminth-infected lungs. Fibroblasts, and likely differentiated myofibroblasts, respond to TGF-β signaling by inducing IL-11 expression, consistent with established profibrotic pathways. In contrast, the mechanisms by which larval stages directly stimulate epithelial, endothelial, and smooth muscle cells remain unclear. Here, we hypothesize that these structural cell populations act as early responders to tissue damage, rapidly upregulating IL-11 and positioning it as an alarmin-like mediator that amplifies downstream inflammatory responses during acute infection. IL-11 signaling within epithelial and stromal compartments promotes the production of neutrophil-associated mediators, including CXCL-1, G-CSF, and IL-6, leading to an early and robust recruitment of neutrophils from the circulation into lung tissue. Recruited neutrophils, themselves a major source of IL-6, further contribute to sustained pulmonary inflammation. Notably, IL-11-producing cell populations express high levels of IL-11Rα1, supporting a model in which IL-11 signaling is reinforced through autocrine and paracrine feedback loops within damaged lung niches during helminth infection. Figure created with BioRender.

Article Snippet: After incubation, mouse IL-11 levels were measured in the culture supernatant by using the Duoset ELISA Kit (R&D Systems).

Techniques: Migration, Infection, Expressing