mouse 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: iScience

Article Title: TLR ligand sensing by lymph node FRCs directs intranodal lymphocyte accumulation to promote immune responses

doi: 10.1016/j.isci.2025.113734

Figure Lengend Snippet: LPS directs intranodal lymphocyte accumulation via increased CCL19/CCL21 expression (A–C) Lymphocyte numbers in the popliteal LNs of WT (A), Tlr4 −/− (B) or Myd88 −/− (C) mice 6 h after s.c. injection of PBS or LPS. n = 12 WT mice/group, 9–10 Tlr4 −/− mice/group, 6–8 Myd88 −/− mice/group. Unpaired t test. Numbers at the top of the graphs indicate fold changes in cellularity as compared to PBS injections. (D) Representative FACS plots, number of de novo accumulating lymphocytes, and competitive B and T cell ratios between CD45.1 + WT and CD45.2 + Tlr4 −/− lymphocytes recovered from the popliteal lymph nodes of CD45.1 + CD45.2 + WT recipients 6 h after i.v cell injection, and PBS or LPS s.c administration. n = 7 mice/group. two-way ANOVA with Sidak’s multiple comparisons post-test (left) and unpaired t test (right). (E) Number of de novo accumulating CD45.1 + WT B cells and T cells recovered from the popliteal lymph nodes of WT: Tlr4 −/− chimeric mice 6 h after i.v cell injection and LPS s.c administration. n = 9 mice/group. one-way ANOVA with Tukey’s multiple comparisons post-test. (F) Representative immunofluorescence images and quantification of intranodal PNAd, CCL21, CXCL13 and VCAM1 expression on lymph node stromal cells, 6 h after PBS or LPS administration to WT mice. Signals are depicted in a divergent scale of red to yellow. Scale bar, 200 μm; insets = 50 μm. n = 4 mice/group. Two-way ANOVA with Sidak’s multiple comparisons post-test. Numbers at the top of the graphs indicate fold changes in MFI expression as compared to PBS injections. (G) Intranodal CCL19, CCL21, and CXCL13 expression as quantified by ELISA, 6 h after PBS or LPS administration to WT, Tlr4 −/− or Myd88 −/− mice. n = 3–4 mice/group. Two-way ANOVA with Sidak’s multiple comparisons post-test. (H) Representative FACS plots and number of de novo accumulating CD45.1 + WT and CD45.2 + Ccr7 −/− lymphocytes recovered from the popliteal lymph nodes of CD45.1 + CD45.2 + WT recipients 6 h after i.v cell injection, and PBS or LPS s.c administration. n = 5 mice/group. Two-way ANOVA with Sidak’s multiple comparisons post-test. (I) Representative FACS plots and number of de novo accumulating CD45.1 + WT lymphocytes recovered from the popliteal lymph nodes of CD45.2 + WT recipients 6 h after i.v donor cell injection, and PBS, recombinant CCL19 or recombinant CCL21 s.c administration. n = 5 mice/group. one-way ANOVA with Dunn’s multiple comparisons post-test. Data represent the mean ± SD with superimposed individual data points. n.s, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.

Article Snippet: In some experiments, lymphocyte migration to and from lymph nodes was manipulated with anti-CD62L antibodies (BioXcell – clone Mel14; 100ug/mouse i.p ) or FTY720 (Cayman Chemicals; 10ug/mouse i.p ): in others experiments, 0.25ug of recombinant CCL19 or recombinant CCL21 proteins (R&D Systems) were injected subcutaneously in the footpad.

Techniques: Expressing, Injection, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Recombinant

Journal: iScience

Article Title: TLR ligand sensing by lymph node FRCs directs intranodal lymphocyte accumulation to promote immune responses

doi: 10.1016/j.isci.2025.113734

Figure Lengend Snippet: TLR4 expression in FRCs supports T cell responses (A) Representative histograms and frequency of TdTomato + cells in Salsa6f fl/+ and Salsa6f fl/+ x Pdgfrb-Cre. ert2 mice, 7 days after the last tamoxifen injection. n = 4–6 mice/group. (B) Representative immunofluorescence images of Salsa6f fl/+ x Pdgfrb-Cre. ert2 lymph nodes, 7 days after the last tamoxifen injection. Arrows in the top inset highlight examples of TdTomato + T cell zone FRCs, arrows in the middle inset highlight TdTomato + CD34 + capsular cells and, arrows in the bottom inset depict TdTomato + Itga7 + perivascular cells. Scale bars, 200 μm. n = 4 mice. (C) Tlr4 mRNA expression in lymph node stromal cells sorted from the nodes of Tlr4 fl/fl and Tlr4 fl/fl x Pdgfrb-Cre. ert2 mice. n = 3 mice/group. Two-way ANOVA with Fisher’s multiple comparisons post-test. (D) Quantification of GCaMP median fluorescence intensity prior to and after ex vivo LPS stimulation of single cells suspensions of Salsa6f fl/+ x Pdgfrb-Cre. ert2 lymph nodes. n = 4 mice; two-way ANOVA with Fisher’s LSD test. (E) Endogenous lymphocyte numbers in non-LPS exposed auricular LNs of tamoxifen-treated Tlr4 fl/fl and Tlr4 fl/fl x Pdgfrb-Cre. ert2 mice. n = 6–8 mice/group. Unpaired t test. (F) Number of de novo accumulating CD45.1 + WT lymphocytes recovered from the non-LPS exposed auricular lymph nodes of tamoxifen-treated Tlr4 fl/fl and Tlr4 fl/fl x Pdgfrb-Cre. ert2 recipients 6 h after i.v cell injection. n = 6–8 mice/group. Unpaired t test. (G) Number of de novo accumulating CD45.1 + WT lymphocytes recovered from the LPS exposed popliteal lymph nodes of tamoxifen-treated Tlr4 fl/fl and Tlr4 fl/fl x Pdgfrb-Cre. ert2 recipients 6 h after i.v cell injection, and LPS s.c administration. n = 6–8 mice/group. Unpaired t test. Numbers at the top of the graphs indicate fold changes in intranodal cellularity. (H) Intranodal CCL19, CCL21, and CXCL13 expression as quantified by ELISA, 6 h after PBS or LPS administration to tamoxifen-treated Tlr4 fl/fl or Tlr4 fl/fl x Pdgfrb-Cre. ert2 mice. n = 4 mice/group. Two-way ANOVA with Sidak’s multiple comparisons post-test. (I) Representative immunofluorescence images and quantification of intranodal CCL21 on lymph node stroma cells in the popliteal lymph nodes of tamoxifen-treated Tlr4 fl/fl and Tlr4 fl/fl x Pdgfrb-Cre. ert2 mice 6 h after LPS administration. CCL21 signal is depicted in a divergent scale of red to yellow. Scale bars, 100 μm. n = 3 mice/group. two-way ANOVA with Sidak’s multiple comparisons post-test. (J and K) Representative division profiles (CTV dilution), number of OT-I (J) and OT-II (K) TCR transgenic cells and their proliferation metrics following recovery from the popliteal lymph nodes of tamoxifen-treated Tlr4 fl/fl and Tlr4 fl/fl x Pdgfrb-Cre. ert2 mice 3 days after i.v T cell transfer and OVA + LPS s.c administration. n = 7–11 mice/group. Unpaired t test. Numbers at the top of the graphs indicate fold changes in OT-I or OT-II cellularity. Data represent the mean ± SD with superimposed individual data points. n.s, not significant; ∗, p < 0.05; ∗∗, p < 0.01.

Article Snippet: In some experiments, lymphocyte migration to and from lymph nodes was manipulated with anti-CD62L antibodies (BioXcell – clone Mel14; 100ug/mouse i.p ) or FTY720 (Cayman Chemicals; 10ug/mouse i.p ): in others experiments, 0.25ug of recombinant CCL19 or recombinant CCL21 proteins (R&D Systems) were injected subcutaneously in the footpad.

Techniques: Expressing, Injection, Immunofluorescence, Fluorescence, Ex Vivo, Enzyme-linked Immunosorbent Assay, Transgenic Assay