Cystic fibrosis cases missed by newborn bloodspot screening—towards a consistent definition and data acquisition

Repeated European surveys of newborn bloodspot screening (NBS) have shown varied strategies for collecting missed cases, and information on data collection differs among countries/regions, hampering data comparison. The ECFS Neonatal Screening Working Group defined missed cases by NBS as either false negatives, protocol-related, concerning analytical issues, or non-protocol-related, concerning pre- and post-analytical issues. A questionnaire has been designed and sent to all key workers identified in each NBS programme to assess the feasibility of collecting data on missed cases, the stage of the NBS programme when the system failed, and individual patient data on each missed case.</p

Airway leukocyte number following LESB65 infection.

<p>Average B lymphocyte, CD4+ve T cell, CD8+ve T cell and neutrophil count shown as mean and standard deviation per mg of tissue at 24, 48, and 72 hours post infection [N = 5 each time point]. * = p<0.05, ** = p<0.01.</p

BAFF expression in BAL fluid.

<p>Expression was measured by ELISA in BAL fluid from <i>P. aeruginosa</i> positive [n = 6] and negative [n = 14] CF patients and healthy controls [n = 7]. BAFF expression was significantly elevated [p<0.01] in both the PA infected group and non PA infected group in comparison to the control group.</p

Expression of BAFF, CXCL13, CCL19 and CCL21 in mouse lung tissue.

<p>Each cytokine was measured at days 1, 2, 3 and 7 days after infection with 10⁶ CFU of <i>P. aeruginosa</i> LESB65 by ELISA. [n = 5]. BAFF was significantly expressed at days 1 [p<0.001], 2 [p<0.01], 3 [p<0.001], and 7 [p<0.05] in comparison to day 0 control. CXCL13 at days 1, 2, 3, [all p<0.001] and 7 [p<0.01]. CCL19 at days 1 [p<0.01], 2 [p<0.001], and 7 [p<0.001]. CCL21 at days 2 [p<0.001], 3 [p<0.01] and 7 [p<0.001]. * = p<0.05, ** = p<0.01, *** = p<0.001.</p

Kinetics of airway infection by <i>P. aeruginosa</i> LESB65.

<p>Mice were challenged [intranasal] with 10⁶ CFU. Values shown as an average Log CFU/mg of lung tissue homogenised, n = 5 mice at each time point. Time zero samples were taken at 15 minutes post infection to confirm entry into the lung and determine levels immediately post infection.</p

FACS analysis of lung inflammatory cells following <i>P. aeruginosa</i> LESB65 infection of mice.

<p>Representative FACS plots showing immune cell staining of airway cells. Each column shows plots from an individual mouse at each time point. All plots are lung CD45+ cells and gates were drawn relative to isotype control antibody staining for each marker. Top row shows B cells [CD45+, CD19+], middle row shows CD4 T cells [CD45+, CD3+, CD4+] and CD8 T cells [CD45+, CD3+, CD8+], bottom row shows neutrophils [CD4+, Gr-1hi, F4/80-] and monocytes/macrophages [CD45+, F4/80+, Gr-1int/low]. Numbers on gates are percentages of total CD45+ cells.</p