Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: Work panel used and monoclonal antibody/dye information.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques:

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: AEA patient's non-activated CD4 + T cells show increased surface expression of CD46, which is dependent on pollen season. Isolated CD4 + T cells from 49 HDs and 58 patients with AEA were analyzed non-activated (NA). The surface expression of CD46 on NA CD4 + T cells was assessed in HDs, AEA in LPP and HPP (A) as well as in HDs vs. AEA based on disease severity in LPP (B) and HPP (C) . Data were measured by flow cytometry and are presented as median fluorescence intensity (MFI) in graphs (A–C) . Horizontal bars represent the median. Next, the surface expression of CD46 was analyzed in HDs and AEA CD4 + T cells under both non-activated (NA) conditions (D) and after stimulation with αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and a high dose of IL-2 (50 U/ml), depending on LPP and HPP (E) . Concentration of sCD46 was assessed in plasma samples using ELISA in 40 HDs and 40 AEA patients in both LPP and HPP (F) . Data are presented as a median +95% CI in graphs (D) , (E) and (F) . Statistical analysis was performed using the unpaired Mann–Whitney U -test for two-group comparisons, and the Kruskal–Wallis test with Dunn's correction for three or more comparisons within a single graph where appropriate; ns (not significant), * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. HDs, healthy donors; AEA, allergic eosinophilic asthma; NA, non-activated; LPP, low pollen period; HPP, high pollen period; MFI, median fluorescence intensity; CI, confidence interval.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Expressing, Isolation, Flow Cytometry, Fluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: AEA patient's CD4 + T cells show increased expression of CD46 after stimulation in low pollen period. CD4 + T cells from 49 HDs and 58 patients with AEA were cultured with a mixture of αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and high dose of IL-2 (50 U/ml) (αCD3/αCD46/IL-2) for 60 h. (A) Surface expression of CD46 on CD4 + T cells was compared in both HDs and AEA patients in dependence of LPP and HPP, as well as between AEA patients in dependence of LPP and HPP (B) . Serum ECP levels were correlated with percentage of CD46 + CD4 + T cells after αCD3/αCD46/IL-2 stimulation among HDs, AEA LPP and AEA HPP (C) . Surface expression of CD46 was measured by flow cytometry and is presented as median fluorescence intensity (MFI). Statistical analysis of CD46 expression was performed using Kruskal–Wallis test with Dunn's correction for multiple comparisons all vs. all. Correlation analysis was performed using non-parametric Spearman correlation coefficient; ns (not significant), * p ≤ 0.05, ** p ≤ 0.01. HDs, healthy donors; AEA, allergic eosinophilic asthma; mAbs, monoclonal antibodies; LPP, low pollen period; HPP, high pollen period; ECP, eosinophilic cationic protein; MFI, median fluorescence intensity.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Expressing, Cell Culture, Flow Cytometry, Fluorescence

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: CD4 + T cells differ in CD46 downregulation after calcitriol co-stimulation in both HDs and AEA patients. CD4 + T cells from 49 HDs and 58 patients with AEA were cultured without stimuli (NA) or with a mixture of αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and high dose of IL-2 (50 U/ml) (αCD3/αCD46) for 60 h alone or with calcitriol (1 × 10 −7 M). The surface expression of CD46 was analyzed before/after calcitriol stimulation in HDs (A) and AEA patients in LPP (B) . Subsequently, two different groups were identified based on CD46 expression after calcitriol stimulation based on following formula: x = MFI CD46 (αCD3/αCD46/IL-2/Calcitriol)—MFI CD46 (αCD3/αCD46/IL-2). When x < 0 = Group CD46D (Decrease) (A1,B1) , when x > 0 = Group CD46I (Increase) (A2,B2) . Representative histograms depicts surface CD46 expression on NA or activated CD4 + T cells in groups CD46D/CD46I in HDs (C) and AEA patients (D) . Statistical analysis was performed using Kruskal–Wallis test with Dunn's correction for multiple comparisons all vs. all; ns (not significant), * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. HDs, healthy donors; AEA, allergic eosinophilic asthma; NA, non-activated; mAbs, monoclonal antibodies; LPP, low pollen period; CD46D, group where CD46 expression on CD4 + T cells decrease after calcitriol stimulation; CD46I, group where CD46 expression on CD4 + T cells increase after calcitriol stimulation.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Cell Culture, Expressing

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: CD46D and CD46I groups of HDs and AEA patients differ in CD46 expression on both non-activated and stimulated CD4 + T cells. CD4 + T cells from 49 HDs and 58 patients with AEA were cultured without stimuli (NA) or with a mixture of αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and high dose of IL-2 (50 U/ml) (αCD3/αCD46/IL-2) and with calcitriol (1 × 10 −7 M) (αCD3/αCD46/IL-2/Cal). Surface expression of CD46 was measured in both groups CD46D/CD46I from HDs (A) , as well as from AEA patients (B) . Subsequently, percentage of CD46 downregulation after αCD3/αCD46/IL-2/Calcitriol stimulation was assessed in the group CD46D from HDs and AEA patients (C) . Identically, percentage of CD46 upregulation after αCD3/αCD46/IL-2/Calcitriol stimulation was assessed in the CD46I group from HDs and AEA patients (D) . The surface expression of CD46 was measured by flow cytometry and is presented as median fluorescence intensity (MFI) in graphs (A) and (B) . Graphs (C) and (D) are depicted as median + 95% CI. Statistical analysis was performed using the non-parametric Mann–Whitney U test; ns (not significant), * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. HDs (healthy donors), AEA, allergic eosinophilic asthma; NA, non-activated; mAbs, monoclonal antibodies; CD46D, group where CD46 expression on CD4 + T cells decrease after calcitriol stimulation; CD46I, group where CD46 expression on CD4 + T cells increase after calcitriol stimulation; MFI, median fluorescence intensity; CI, confidence interval.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Expressing, Cell Culture, Flow Cytometry, Fluorescence, MANN-WHITNEY

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: Increased CD46 expression on CD4 + T cells from both HDs and AEA patients is independent of serum 25-OH vitamin D concentration. To clarify the relationship between the surface expression of CD46 on NA CD4 + T cells and serum levels of 25-OH vitamin D, these parameters were correlated in both HDs ( n = 49) (A) and AEA patients in LPP ( n = 58) (B) , as well as in HPP (C) . Next, serum levels of 25-OH vitamin D were compared between HDs and AEA (D) as well as in dependence on asthma severity (E) . The serum concentration of 25-OH vitamin D was also compared after dividing HDs and AEA patients into CD46D/CD46I groups (F) . Correlation analysis was performed using the non-parametric Spearman correlation coefficient, comparison of serum 25-OH vitamin D levels between HDs and AEA was performed using the non-parametric Mann–Whitney U -test, while the Kruskal–Wallis test with Dunn's correction was used for multiple comparisons all vs. all; ns (not significant), * p ≤ 0.05. HDs, healthy donors; AEA, allergic eosinophilic asthma; NA, non-activated; CD46D, group where CD46 expression on CD4 + T cells decrease after calcitriol co-stimulation; CD46I, group where CD46 expression on CD4 + T cells increase after calcitriol co-stimulation.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Expressing, Concentration Assay, Comparison, MANN-WHITNEY

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: AEA patients in low pollen period show increased CD25 expression on stimulated CD4 + T cells, which is further promoted by calcitriol. CD4 + T cells from 49 HDs and 58 patients with AEA were cultured with a mixture of αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and high dose of IL-2 (50 U/ml) (αCD3/αCD46/IL-2) or with calcitriol (1 × 10 −7 M) (αCD3/αCD46/IL-2/Cal) for 60 h. (A) Surface expression of CD25 (IL-2 receptor α chain) and proliferation (B) were analyzed by flow cytometry before/after calcitriol stimulation with αCD3/αCD46/IL-2 in HDs and AEA patients in LPP and HPP groups using the non-parametric Mann–Whitney U -test. The data are presented as median intensity fluorescence (MFI), where horizontal bars indicate the median. Based on the CD46 dynamics after calcitriol stimulation, HDs and AEA patients were divided into groups CD46D/CD46I and surface expression of CD25 was analyzed separately. (C) To better understand the effect of calcitriol on CD25 expression in CD4 + T cells between HDs and AEA patients, we expressed the results as the percentage of upregulation after stimulation. Next, sIL-2RA levels were assessed in cell culture SNs, with HDs and AEA LPP patients divided into CD46D/CD46I groups (E) and are presented as a median +95% CI. Data from panels (C) and (E) were analyzed using the Wilcoxon-sign rank test and data from graphs (D) and (F) using the Kruskal–Wallis test with Dunn's correction; ns (not significant), * p ≤ 0.05. HDs, healthy donors; AEA, allergic eosinophilic asthma; mAbs, monoclonal antibodies; LPP, low pollen period; sIL-2RA, soluble IL-2 receptor α chain; CD46D, group where CD46 expression on CD4 + T cells decrease after calcitriol stimulation; CD46I, group where CD46 expression on CD4 + T cells increase after calcitriol stimulation; MFI, median fluorescence intensity; CI, confidence interval.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Expressing, Cell Culture, Flow Cytometry, MANN-WHITNEY, Fluorescence

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: Stimulated CD4 + T cells from CD46D group produce significantly more IFN-γ and IL-10 in HDs and react to calcitriol co-stimulation with downregulation of IFN-γ and upregulation of IL-10. CD4 + T cells from 49 HDs were cultured with a mixture of αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and high dose of IL-2 (50 U/ml) (αCD3/αCD46/IL-2) or with calcitriol (1 × 10 −7 M) (αCD3/αCD46/IL-2/Cal) for 60 h. The concentrations of IFN-γ (A) and IL-10 (B) were assessed using ELISA and analyzed in accordance with the division of HDs into CD46D/CD46I groups. The results are presented as the median +95% CI. Similarly, the percentages of IFN-γ + CD4 + T cells (C) and IL-10 + CD4 + T cells (D) obtained from flow cytometry were analyzed, with the horizontal bar representing the median. Data were analyzed using the Kruskal–Wallis test with Dunn's correction for multiple comparisons all vs. all. (E) The timeline showing IFN-γ and IL-10 production by αCD3/αCD46/IL-2-stimulated CD4 + T cells at 0, 12, 36, 60 and 84 h of incubation is based on data from three healthy donors and is presented as the median + 95% confidence interval. (F) Representative dot-plots depict differences in IFN-γ and IL-10 production by activated CD4 + T cells respecting the division into the groups CD46D/CD46I. The data from flow cytometry were obtained from CD4 + T cells treated with Brefeldin A during the final 4 h of stimulation and were gated from 90.000 CD4 + T cells. The numbers in the top right corner show percentage of CD4 + T cells in each quadrant. The data are representatives of 107 individual samples performed in 18 series. HDs, healthy donors; mAbs, monoclonal antibodies; CD46D, group where CD46 expression on CD4 + T cells decrease after calcitriol stimulation; CD46I, group where CD46 expression on CD4 + T cells increase after calcitriol stimulation.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Incubation, Expressing

Journal: Frontiers in Allergy

Article Title: Low CD46 expression on activated CD4 + T cells predict improved Th1 cell reactivity to calcitriol in majority of patients with allergic eosinophilic asthma and healthy donors

doi: 10.3389/falgy.2024.1462579

Figure Lengend Snippet: AEA patient's CD4 + T cells from CD46D group show increased production of IFN-γ after stimulation, which can be downregulated by calcitriol and simultaneously produce more IL-10. CD4 + T cells from 49 HDs and 58 AEA patients were cultured with a mixture of αCD3 (10 μg/ml), αCD46 (5 μg/ml) mAbs and high dose of IL-2 (50 U/ml) (αCD3/ α CD46/IL-2) or with calcitriol (1 × 10 −7 M) (αCD3/αCD46/IL-2/Cal) for 60 h. Concentrations of IFN-γ and IL-10 were measured in cell culture SNs using ELISA. IFN-γ production was compared between AEA LPP and AEA HPP groups (A) as well as in groups CD46D (B) and CD46I (C) . An identical analysis was performed with production of IL-10 (D–F) . To simplify the evaluation of the calcitriol's effect in both HDs and AEA patients, with respect to the CD46D/CD46I group division, results are presented as the percentage of downregulation in IFN-γ (G) and percentage of upregulation in IL-10 (H) after αCD3/αCD46/IL-2/Cal stimulation. Results are presented as the median + 95% CI. Paired data were analyzed using Wilcoxon matched-pairs signed rank test (black color) and unpaired data were analyzed unpaired Mann–Whitney U -test (grey color); ns (not significant), * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. HDs, healthy donors; AEA, allergic eosinophilic asthma; mAbs, monoclonal antibodies; SNs, supernatant; CD46D, group where CD46 expression on CD4 + T cells decrease after calcitriol stimulation; CD46I, group where CD46 expression on CD4 + T cells increase after calcitriol stimulation; CI, confidence interval.

Article Snippet: CD46 , AF700 , MEM-258 , A7-342-T100 , Exbio.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Expressing

Journal:

Article Title: Treatment of murine lupus with cDNA encoding IFN-?R/Fc

doi:

Figure Lengend Snippet: IgG deposits and expression of MHC class II, ICAM-1, and MCP-1 proteins in kidney tissues from MRL-Faslpr mice treated from 1 month of age with blank (VR1255) or active (VR1255-IFNγR/Fc) plasmid with electroporation. Active plasmid–treated mice had substantially reduced glomerular IgG deposits, as well as decreased levels of MHC class II, ICAM-1, and MCP-1. Similar reductions were observed in mice treated from 4 months of age with active plasmid (data not shown). IgG deposits were detected with FITC–anti-IgG antibody (×45), and the immunoperoxidase procedure for detection of MHC class II, ICAM-1, and MCP-1 was performed as described in Figure ​Figure55 (×25). Photomicrographs are representative samples from four mice per group.

Article Snippet: Subsequently, sections were incubated with various primary antibodies, including anti–IgG-FITC (Vector Labs), anti-CD3-biotin (PharMingen), anti–F4/80-biotin (Caltag), anti–ICAM-1 (PharMingen), anti–MHC class II (PharMingen), and anti–MCP-1 (Santa Cruz Biotechnology, Santa Cruz, California, USA).

Techniques: Expressing, Plasmid Preparation, Electroporation

Journal:

Article Title: Protection against inflammation- and autoantibody-caused fetal loss by the chemokine decoy receptor D6

doi: 10.1073/pnas.0607514104

Figure Lengend Snippet: Chemokines in the LPS model of fetal loss. (A–E) Serum chemokine concentrations after LPS treatment in WT and D6−/− male mice. WT (open symbols) and D6−/− (filled symbols) mice were injected i.p. with 1.35 mg/kg LPS. At the indicated time points, circulating chemokine concentrations were measured by ELISA. Data are from seven mice for each time point. (F–J) Serum chemokine concentrations. WT (open columns) and D6−/− (filled columns) mice at day 10 of pregnancy were injected i.p. with 0.4 mg/kg LPS. Circulating chemokine concentrations were measured at 8 h postinjection by ELISA. Data are from nine WT and eight D6−/− mice. (K–O) Chemokine levels in placenta. WT (open columns) and D6−/− (filled columns) mice at day 10 of pregnancy were injected i.p. with 0.4 mg/kg LPS. Chemokine concentrations (expressed as nanograms of chemokine per milligram of total proteins of the lysates) were measured at 8 h postinjection by ELISA. Data are from nine WT and eight D6−/− mice. Results are reported as mean ± SEM. (A, F, and K) CCL22. (B, G, and L) CCL2. (C, H, and M) CCL11. (D, I, and N) CCL3. (E, J, and O) CXCL2.

Article Snippet: To block inflammatory chemokines, animals were treated with a mixture of goat antibodies to the mouse CC chemokines CCL3L1 (catalog no. AB450NA), CCL4 (catalog no. AB451NA), CCL2 (catalog no. AB479NA) and rat mAb anti-mouse CCL5 (catalog no. MAB478) purchased lyophilized from R&D Systems, resuspended in PBS, and mixed.

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Whole-lung murine IL-13, CCL6, and CCL21 levels in C.B-17SCID/bg mice that received normal, NSIP, or IPF/UIP fibroblasts. Lung samples were removed at days 35 and 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice, and all soluble proteins were measured by specific ELISA. Data shown are mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 compared with appropriate the C.B-17SCID/bg group that received normal fibroblasts. ττP ≤ 0.01, τττP ≤ 0.001 compared with whole-lung cytokine and chemokine levels at the day 35 time point in the C.B-17SCID/bg groups with either IPF/UIP or NSIP fibroblasts The whole-lung cytokine and chemokine levels in control C.B-17SCID/bg group that did not receive fibroblasts were as follows: IL-13, 0.18 ± 0.014 ng/mg protein; CCL6, 0.37 ± 0.05 ng/mg protein; and CCL21, 2.0 ± 0.1 ng/mg protein.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: SuperArray Analysis of Human CCR7 and CCL21 and TaqMan Analysis of Human CCR7 in Whole-Lung Samples from C.B-17SCID/bg Mice at Day 35 after i.v. Human Fibroblast Injection

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Injection

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Representative Mason trichrome-stained histological sections from C.B-17SCID/bg mice that received normal (A–C), NSIP (D–F), or IPF/UIP (G–I) fibroblasts. No interstitial remodeling was apparent in C.B-17SCID/bg mice that received normal fibroblasts, but vascular anomalies were observed in this group (B), and the IgG (A), anti-CCL21 monoclonal antibody (B), and anti-CCR7 monoclonal antibody (C) therapies did not alter the lung histological appearance in this group. Pulmonary remodeling was apparent in C.B-17SCID/bg mice that received NSIP fibroblasts, and this pattern was not altered by IgG (D), whereas the anti-CCL21 antibody (E) or anti-CCR7 antibody (F) therapies markedly reduced the interstitial remodeling in whole-lung samples. Interstitial pulmonary fibrosis was apparent in C.B-17SCID/bg mice that received IPF/UIP fibroblasts, and this pattern was not altered by IgG (G), whereas the anti-CCL21 antibody (H) or anti-CCR7 antibody (I) therapies markedly reduced the interstitial remodeling in whole-lung samples. Monoclonal antibody therapies began at day 35 and continued to day 63, and lung samples were removed at day 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice. Original magnification, ×400.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Staining

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Quantitative TaqMan PCR analysis of extracellular matrix-associated genes MMP-2 (top) and MMP-19 (bottom) in C.B-17SCID/bg mice that received normal, NSIP, or IPF/UIP fibroblasts. Changes in gene expression are expressed as mean ± SEM of the fold increase in transcript expression above a group of C.B-17SCID/bg mice that received PBS, PKH26, and one of IgG, anti-CCL21 antibody, and anti-CCR7 antibody. *P ≤ 0.05, ***P ≤ 0.001 compared with the appropriate C.B-17SCID/bg group that received human fibroblasts and IgG treatment.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Expressing

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Whole-lung hydroxyproline levels in C.B-17SCID/bg mice that received no fibroblasts (ie, control) or received normal, NSIP, or IPF/UIP fibroblasts. All groups of mice received either IgG or monoclonal antibody therapy. IgG, anti-CCL21, and anti-CCR7 monoclonal antibody therapies began in separate groups of C.B-17SCID/bg mice at day 35 and continued to day 63. Lung samples were removed at day 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice. Data shown are mean ± SEM. *P ≤ 0.05 compared with the control C.B-17SCID/bg group, which did not receive any human fibroblasts; τττP ≤ 0.001 compared with the appropriate C.B-17SCID/bg group that received human fibroblasts and IgG treatment; τP ≤ 0.05 compared with the appropriate C.B-17SCID/bg group that received human fibroblasts and IgC treatment.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Control

Journal:

Article Title: Therapeutic Targeting of CC Ligand 21 or CC Chemokine Receptor 7 Abrogates Pulmonary Fibrosis Induced by the Adoptive Transfer of Human Pulmonary Fibroblasts to Immunodeficient Mice

doi: 10.2353/ajpath.2007.060649

Figure Lengend Snippet: Whole-lung murine IL-13, CCL6, and CCL21 levels in C.B-17SCID/bg mice that received normal, NSIP, or IPF/UIP fibroblasts and IgG or monoclonal antibody therapies. Anti-CCL21 and anti-CCR7 monoclonal antibody therapies began at day 35 and continued to day 63. Lung samples were removed at day 63 after the adoptive i.v. transfer of human fibroblasts into C.B-17SCID/bg mice. Specific ELISA was used to measure all soluble proteins. Data shown are mean ± SEM. *P ≤ 0.05, **P ≤ 0.01 compared with indicated protein levels measured in whole-lung samples from C.B-17SCID/bg mice that received human fibroblasts.

Article Snippet: Thirty-five days later, all groups of five C.B-17SCID/bg mice received mouse IgG, mouse anti-human CCL21 monoclonal antibody, or mouse anti-human CCR7 monoclonal antibody (all at 10 μg/ml; R&D Systems, Minneapolis, MN) every other day from days 35 to 63.

Techniques: Enzyme-linked Immunosorbent Assay