Reagents and antibodies for Tfh panel.

Reagents and antibodies for Tfh panel.</p

Age related changes in selected T cell populations.

A, B) A decrease in naïve CD4+ and CD8+ T cells with advancing age (C, D) corresponds with an increase in central memory CD4+ and CD8+ T cells. E) Effector memory CCR4+ Tregs increase with age. Tregs, identified by gating on CD4+CD25+CD127low cells were compared based on four age groups.</p

Racial disparities in the distribution of Tfh cells.

A) Overall Tfh frequencies were similar between African-Americans and Caucasians. However, race related differences were observed in certain Tfh cell subsets including B, C) decreased frequencies of Tfh1 cells in African-Americans at all age ranges, and D, E) increased Tfh17 frequencies in African-Americans, a difference that was observed in all age groups. In C and E, Caucasians are represented by blue bars and African-Americans by orange bars.</p

Quantitative Proteomics of Bronchoalveolar Lavage Fluid in Idiopathic Pulmonary Fibrosis

The proteomic analysis of bronchoalveolar lavage fluid (BALF) can give insight into pulmonary disease pathology and response to therapy. Here, we describe the first gel-free quantitative analysis of BALF in idiopathic pulmonary fibrosis (IPF), a chronic and fatal scarring lung disease. We utilized two-dimensional reversed-phase liquid chromatography and ion-mobility-assisted data-independent acquisition (HDMSE) for quantitation of >1000 proteins in immunodepleted BALF from the right middle and lower lobes of normal controls and patients with IPF. Among the analytes that were increased in IPF were well-described mediators of pulmonary fibrosis (osteopontin, MMP7, CXCL7, CCL18), eosinophil- and neutrophil-derived proteins, and proteins associated with fibroblast foci. For additional discovery and targeted validation, BALF was also screened by multiple reaction monitoring (MRM), using the JPT Cytokine SpikeMix library of >400 stable isotope-labeled peptides. A refined MRM assay confirmed the robust expression of osteopontin, and demonstrated, for the first time, upregulation of the pro-fibrotic cytokine, CCL24, in BALF in IPF. These results show the utility of BALF proteomics for the molecular profiling of fibrotic lung diseases and the targeted quantitation of soluble markers of IPF. More generally, this study addresses critical quality control measures that should be widely applicable to BALF profiling in pulmonary disease

Quantitative Proteomics of Bronchoalveolar Lavage Fluid in Idiopathic Pulmonary Fibrosis

The proteomic analysis of bronchoalveolar lavage fluid (BALF) can give insight into pulmonary disease pathology and response to therapy. Here, we describe the first gel-free quantitative analysis of BALF in idiopathic pulmonary fibrosis (IPF), a chronic and fatal scarring lung disease. We utilized two-dimensional reversed-phase liquid chromatography and ion-mobility-assisted data-independent acquisition (HDMSE) for quantitation of >1000 proteins in immunodepleted BALF from the right middle and lower lobes of normal controls and patients with IPF. Among the analytes that were increased in IPF were well-described mediators of pulmonary fibrosis (osteopontin, MMP7, CXCL7, CCL18), eosinophil- and neutrophil-derived proteins, and proteins associated with fibroblast foci. For additional discovery and targeted validation, BALF was also screened by multiple reaction monitoring (MRM), using the JPT Cytokine SpikeMix library of >400 stable isotope-labeled peptides. A refined MRM assay confirmed the robust expression of osteopontin, and demonstrated, for the first time, upregulation of the pro-fibrotic cytokine, CCL24, in BALF in IPF. These results show the utility of BALF proteomics for the molecular profiling of fibrotic lung diseases and the targeted quantitation of soluble markers of IPF. More generally, this study addresses critical quality control measures that should be widely applicable to BALF profiling in pulmonary disease

Example of web-based data visualization tool.

Researchers have the ability to select reference ranges for flow cytometry panels of interest by age, gender, and race filters. This tool will calculate summary statistics on the selected population and the data may be downloaded for study. Links to cell ontology are present when applicable, and the gating strategies are available. Gating strategies and summary tables are also available.</p

Study population demographics.

Study population demographics.</p

Forest plot summarizing reference ranges for major lymphocyte subsets in the overall population.

References are shown for CD3+ T cells, CD4+ T cells, CD8+ T cells, Tfh cells, overall B cells, monocytes, NK cells, Tregs, and dendritic cells. Data are presented as means with 95% reference intervals calculated by non-parametric bootstrap.</p

Reagents and antibodies for HIPC panels.

Reagents and antibodies for HIPC panels.</p

Racial disparities in the distribution of innate cells.

A) Classical (CD14+CD16-) monocytes were decreased and B) non-classical (CD16+CD14-) were increased in African-Americans compared with Caucasians. C) The increase in CD16+CD14- populations in African-Americans was present in all age groups. D) In each age group, the frequency of CD14+ monocytes was higher in Caucasians compared to African-Americans. E) CD16+CD56- NK cell frequencies were increased in African-Americans compared to Caucasians, and the difference became larger in the older age groups. In C, D, and E, Caucasians are represented by blue bars and African-Americans by orange bars.</p