Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: Chemokines and antibodies used in SDS/PAGE and Western blot analysis.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Western Blot, Concentration Assay, Recombinant

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: CCL21 expression in systemic sclerosis with associated pulmonary arterial hypertension from explanted lung tissue. CCL21 immunohistochemistry demonstrated protein expression (black arrows) from (A-C) airway epithelial cells (20x) and (D) alveolar macrophages (20x), which was confirmed by (E) CD68 macrophage marker in a serial cut of the same histopathology block.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Expressing, Immunohistochemistry, Marker, Histopathology, Blocking Assay

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: CCL21 serum levels and correlation analysis in ELISA 1, Luminex and ELISA 2. (A) The mean serum levels of CCL21 in ELISA 1 were higher in SSc patients compared to healthy controls (i) in Luminex (ii) and ELISA 2 (iii). (B) In ELISA 1, CCL21 serum levels were higher in patients diagnosed with PAH compared to patients with no PH (i) while similar in PAH patient and no PH (ii) in Luminex. The mean serum levels of CCL21 in ELISA 2 however were, like in ELISA 1, higher in PAH patients compared to those without PH (iii). (C) The correlation of CCL21 serum levels between ELISA 1 and Luminex (r=0.515, p<0.001) (i) and the correlation of CCL21 serum levels between ELISA 2 and Luminex (r=0.52, p<0.001) (ii) were poor. However, the correlation of CCL21 serum levels between ELISA 2 and ELISA 1 (r=0.788, p<0.001) (iii) were good.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Enzyme-linked Immunosorbent Assay, Luminex

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: ROC analysis of CCl21 levels from ELISA 1, Luminex and ELISA 2. (A) ROC analysis of CCL21 levels from ELISA 1 with AUC=0.843, 95% CI = 0.084-0.883 and p<0.001, (B) ROC analysis of CCL21 levels from Luminex with AUC=0.772, 95% CI = 0.711-0.833 and p<0.001, and (C) ROC analysis of CCL21 levels from ELISA 2 with AUC=0.770, 95% CI = 0.711-0.830 and p<0.001.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Enzyme-linked Immunosorbent Assay, Luminex

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: Peptides detected by MS of CCL21 immunoprecipitated from SSc patient serum.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Immunoprecipitation

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: Multivariable logistic and cox regression analysis showing CCL21 and other variables associated with PAH and predictive of PAH.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques:

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: Serum anti-CCL21 antibody level in healthy controls and patients with different systemic connective tissue diseases. The dots in healthy controls (green), Systemic sclerosis (SSc; pink), primary Sjøgren syndrome (SS; purple), Systemic lupus erythematosus (SLE; blue) and Mixed connective tissue disease (MCTD; orange) shows the level of anti-CCL21 in the different patients. The dotted line represents the cut off value for considering the patients positive or negative for anti-CCL21, in which the patients above the dotted line are positive for anti-CCL21 antibodies. Levels of anti-CCL21 antibodies were significantly higher in patients with SSc, SS and SLE than in healthy controls (p-value).

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques:

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: CCL21 levels in SSc, SS and MCTD of anti-CCL21 antibody positive and negative patients. (A) Mean serum levels of CCL21 in anti-CCL21 antibody positive (green) and anti-CCL21 antibody negative (red) patients in SSc, SS and MCTD. (B) Mean serum levels of CCL21 of anti-CCL21 antibody positive (green) and anti-CCL21 antibody negative (red) patients in SSc patients diagnosed with PAH.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques:

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: Test of CCL21-antibodies using Western blot analysis. The Western blot analysis showed that three of the CCL21 antibodies detected both full length and tailless CCL21 (four left lanes). The custom-made anti-C-terminal tail of CCL21 antibody from Sicgen only detected full length CCL21 (right lane).

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

doi: 10.3389/fimmu.2022.991743

Figure Lengend Snippet: Western blot of SSc serum samples. The four first lanes show the IP product of 1: costume made CCL21, 2: Tailless CCL21, 3: Patient serum 1, and 4: patient serum 2. The last two lanes CCL21 and Tailless CCL21 directly loaded on the gel.

Article Snippet: Additionally, and as expected, we found that the custom-made anti-C-terminal CCL21 antibody from Sicgen detected the full length CCL21 protein, but not tailless CCL21 ( ).

Techniques: Western Blot