human ccl21  (R&D Systems)

Journal: Scientific Reports

Article Title: Immune regulation and lymphangiogenesis by lymphatic endothelial cells in the decidua in severe preeclampsia

doi: 10.1038/s41598-026-35667-3

Figure Lengend Snippet: Reduced CCL21 expression and impaired dendritic cell (DC) migration in PE-dLECs. ( A ) PE-dLECs showed significantly reduced expression of CCL21 at both the mRNA and protein levels, as assessed using RT-qPCR and western blot, respectively. ( B ) DC trafficking through LECs was evaluated in three steps: migration, adhesion, and transmigration. PE-dLECs exhibited reduced activity in all three steps. ( C ) Decreased DC migration observed with conditioned media from PE-dLECs was restored by the addition of recombinant CCL21. Data are presented as mean ± SEM. Statistical significance was assessed using one-way ANOVA followed by Tukey’s test. All experiments represent three independent experiments, each performed in duplicate. **, p < 0.01 vs. N-dLEC. N-dLECs, dLECs from normal pregnancies; PE-dLECs, dLECs from preeclamptic pregnancies; DC, dendritic cell; dLECs, decidua lymphatic endothelial cells; PE, preeclampsia; CCL 21, chemokine (C-C motif) ligand 21; RT-qPCR, quantitative reverse-transcription PCR.

Article Snippet: Conditioned media from PE or normal pregnancies collected after 24 h of culture in EBM with 1% FBS were added to the lower chamber, with or without recombinant human CCL21 (250 ng/mL; R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Migration, Quantitative RT-PCR, Western Blot, Transmigration Assay, Activity Assay, Recombinant, Reverse Transcription

Journal: Scientific Reports

Article Title: Immune regulation and lymphangiogenesis by lymphatic endothelial cells in the decidua in severe preeclampsia

doi: 10.1038/s41598-026-35667-3

Figure Lengend Snippet: Proposed model of immune and lymphatic dysfunction at the maternal-fetal interface in preeclampsia. This schematic illustrates the functional and genetic differences observed in PE-dLECs. In PE-dLECs, reduced expression of lymphangiogenesis-related genes leads to impaired lymphatic vessel formation and functional deficits. Decreased CCL21 expression disrupts DC trafficking, while reduced NO levels impair the suppression of cytotoxic T cells. Finally, these changes may contribute to immune imbalance and the pathogenesis of preeclampsia. LV, lymphatic vessel; BV, blood vessel; M, amnion chorion; D, decidua; dLECs, decidual lymphatic endothelial cells; PE, pre-eclampsia; NO, nitric oxide; CCL 21, chemokine (C-C motif) ligand 21; DC, dendritic cell.

Article Snippet: Conditioned media from PE or normal pregnancies collected after 24 h of culture in EBM with 1% FBS were added to the lower chamber, with or without recombinant human CCL21 (250 ng/mL; R&D Systems, Minneapolis, MN, USA).

Techniques: Functional Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Biomaterial-assisted local oxygenation safeguards the prostimulatory phenotype and functions of human dendritic cells in hypoxia

doi: 10.3389/fimmu.2023.1278397

Figure Lengend Snippet: O 2 -cryogels mitigate the exposure of MDDCs to hypoxia. (A) Graphical abstract depicting local oxygenation from O 2 -cryogels preserves the proinflammatory function of DCs. Immature DCs identify and internalize pathogenic antigens via PRRs, initiating antigen processing and presentation via MHC molecules. This process is accompanied by DC maturation and upregulation of activation and co-stimulatory molecules, including CD40, CD86, and CCR7, as well as the secretion of proinflammatory cytokines and chemokines, such as IL-12p70, TNF-α, and MIP-1α. Following this, DCs migrate to draining lymph nodes in response to the chemokine gradient of CCL21, signaling through CCR7 receptor. Here, they prime naive T cells, triggering their activation and proliferation, and thereby mounting an effective immune response. However, the activity of DCs can be substantially compromised under hypoxic conditions found in physiological tissues, solid tumors, and sites of inflammation. Local O 2 supply via O 2 -cryogels can significantly prevent hypoxia-induced inhibition of DC functions, including antigen uptake, maturation, migration, as well as T-cell activation and proliferation, and restore their proinflammatory activity. (B) Profile of O 2 concentration measured in media with and without MDDCs and O 2 -cryogels. MDDCs (1.5x10 5 cells/well) were cultured under hypoxic conditions (1% O 2 ) for 48 h in the presence and absence of O 2 -cryogels, and O 2 tension in the media was monitored using contactless sensor spots. Controls included medium alone, or cells cultured in medium in the presence and absence of O 2 -cryogels. Data are representative of three independent experiments and presented as mean of n = 4 replicates. (C) Time it takes for the MDDCs to become hypoxic in cell culture media in the presence and absence of O 2 -cryogels. Data are representative of three independent experiments and presented as mean ± SD of n = 4 replicates. Statistical analysis was performed using one-way ANOVA and Tukey’s post hoc test using GraphPad software; *P < 0.05, ****P < 0.0001.

Article Snippet: The lower wells were filled with 150 μL media containing 1000 ng/mL recombinant human CCL21 (R&D systems).

Techniques: Activation Assay, Activity Assay, Inhibition, Migration, Concentration Assay, Cell Culture, Software

Journal: Frontiers in Immunology

Article Title: Biomaterial-assisted local oxygenation safeguards the prostimulatory phenotype and functions of human dendritic cells in hypoxia

doi: 10.3389/fimmu.2023.1278397

Figure Lengend Snippet: O 2 -cryogels preserve antigen uptake and chemotaxis function of MDDCs in hypoxia via O 2 and HA-related mechanisms. (A) Schematic of antigen uptake by DCs occurring optimally in normoxia vs. inhibited in hypoxia vs. protected by O 2 -cryogels in hypoxia. MDDCs were preconditioned in either normoxia or hypoxia in cryogel-free medium or medium containing various cryogels (O 2 -cryogel, HAGM cryogel, PEG cryogel) for 24 (h) Then, the cells were exposed to pHrodo Green-labeled OVA and dextran for 1 h, washed, stained for surface markers and MFI in the FITC channel was analyzed using flow cytometry. pHrodo Green exhibits fluorescence in the low pH environment of the lysosome, serving as an indicator of phagocytosis. (B) OVA uptake by MDDCs preconditioned with various cryogels in normoxic and hypoxic conditions. (C) Dextran uptake by MDDCs preconditioned with various cryogels in normoxic and hypoxic conditions. (D) Schematic of migration of DCs occurring optimally in normoxia vs. inhibited in hypoxia vs. maintained by O 2 -cryogels in hypoxia. MDDCs were preconditioned in either normoxia or hypoxia in cryogel-free medium or medium containing various cryogels (O 2 -cryogel, HAGM cryogel, PEG cryogel) for 24h. Then, a transwell-based system was employed to assess the migratory activity of MDDCs. Medium-containing chemokine CCL21 was added in the lower chamber, whereas the preconditioned cells were added on the top chamber. Chemotaxis was allowed to take place for 3 h in hypoxic and normoxic conditions. Subsequently, the migrated cells in the lower chamber were lysed using Lysis Buffer/Cyquant® GR Dye, and the number of migrated cells was measured using fluorescence-based readout. (E) Chemotaxis of MDDCs induced by CCL21 in normoxic and hypoxic conditions post preconditioning with various cryogel formulations. Data are representative of three independent experiments and presented as mean ± SD of n = 4–5 replicates. Statistical analysis was performed using one-way ANOVA and Tukey’s post hoc test using GraphPad software; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: The lower wells were filled with 150 μL media containing 1000 ng/mL recombinant human CCL21 (R&D systems).

Techniques: Chemotaxis Assay, Labeling, Staining, Flow Cytometry, Fluorescence, Migration, Activity Assay, Lysis, CyQUANT Assay, Software