Page 311 - Haematologica April 2020

HSC-GT for herPAP
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Figure 4. Significant improvement of disease-related parameters twelve weeks after Lv.Csf2rb hematopoietic stem cell-based gene therapy (HSC-GT). (A) Representative picture of bronchoalveolar lavage fluid (BALF) presenting clear and see-through fluid in untreated wild-type (WT) and Csf2rb-/- mice transplanted with healthy CD45.1 or Lv.Csf2rb-transduced cells, while Csf2rb-/- mice either untreated or transplanted with Lv.GFP-transduced cells present a hereditary pulmonary alve- olar proteinosis (herPAP) characteristic milky and turbid fluid. (B) BALF turbidity measured as optical density at 600 nm (OD600). Lv.Csf2rb n=5, Lv.GFP n=3, others n=4 in two independent experiments. (C) Total protein concentration (μg/mL) in BALF. Lv.Csf2rb n=5, Lv.GFP n=3, others n=4 in two independent experiments. Cholesterol (μg/mL) (D), granulocyte/macrophage-colony-stimulating factor (GM-CSF) (pg/mL) (E), macrophage colony-stimulating factor (M-CSF) (pg/mL) (F), and monocyte chemoattractant protein-1 (MCP-1) (pg/mL) (G) levels in BALF. Lv.Csf2rb n=5, CD45.1 n=4, others n=3 in two independent experiments. (H) Quantification of Periodic Acid-Schiff (PAS) positive (PAS+) areas in lung slices. Lv.Csf2rb, Lv.GFP and WT n=3, CD45.1 n=2, Csf2rb-/- n= 4. Lines indicate mean±standard error of mean. (I) Representative computed tomography scans during inspiratory (Insp.) and expiratory (Exsp.) phase. (J and K) Quantification of mean lung density in Hounsfield units (HU) measured in computed tomography scans in expiratory (J) and inspiratory (K) phase. Lv.Csf2rb n=6, WT, Csf2rb-/- and Lv.GFP n=3, CD45.1 n=4 in two independent experiments. Lines indicate means. All statistical analysis was carried out by one-way ANOVA with Dunnett’s multiple comparison testing com- paring all samples to untreated Csf2rb-/- mice; ns: not significant; *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.
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