Joselyn Rojas-Quintero 1, Scott A Ochsner 2, Hyun-Sung Lee 6, Christine Cong 1, Alan Waich Cohen 1, Adrianne S Colborg 3, Konstantin Tsoyi 1, Maria C Basil 4, Edward Cantu 5, Ivan O Rosas 1, Neil J McKenna 2, Raúl San-José Estépar 7, Igor Barjaktarevic 8, Andrew A Wilson 9, Francesca Polverino 1,*
Posted: Jan 2, 2025
Abstract
Although the emphysema pathogenesis involves a complex network of gene-environment interactions1, monogenic disease alpha-1-anti-trypsin (AAT) deficiency (AATD) is associated with emphysema. Severe AATD is characterized by the ‘Z’ allele homozygosity in the SERPINA1 gene, where individuals have <20% normal concentrations of AAT protein, predisposing them to liver and lung disease2.
The loss-of-function toxicity based on a protease–antiprotease imbalance has long been the paradigm for the pathogenesis of AATD emphysema. Recent evidence suggests that AAT plays essential roles in B cell regulation, differentiation and activation3. Moreover, AAT augmentation reduces auto-reactive lymphocytic infiltrates in preclinical and clinical autoimmunity4. However, little is known about the contribution of the adaptive immune system to AATD emphysema pathogenesis. Herein, we explored the immune compartment in AATD vs. WT-AAT emphysema using complementary digital spatial profiling (DSP) and immunostaining.
NeoBiotechnologies’ products were used in this study:
Formalin-fixed paraffin-embedded (FFPE) lung sections from 7 ZZ-AATD and 11 wild-type (WT)-AAT patients, who underwent lung transplantation for severe pulmonary emphysema (IRB #H46823, #1811124026, and PROPEL) were used. GeoMx® DSP technology was used to obtain whole transcriptome atlas from four different regions of interest (ROIs): parenchyma, airways, vessels, and lymphoid follicles (LFs) as previously described5 .
Consecutive lung sections were stained by immunofluorescence (IF) for a combination of: AAT protein (ThermoFisher, #711079, Waltham, MA), CD20 (pan-B cell marker, Abcam, #ab219329, Cambridge, MA), CD138 (plasma cell marker, R&D Systems, catalog #AF2780, Minneapolis, MN), CD21 (follicular dendritic cell marker, Abcam, #AB75985-1001), CD10 (transitional B cells, Invitrogen, #PA5-47075, Waltham, MA), CD27 (memory B cell marker, Novus, #NBP2-78837, Centennial, CO), and IgA (mucosal immunoglobulin, NeoBiotechnologies, #3493-MSM1-P1, Union City, CA) to define LF cellular subsets. We also used AAT polymer (HyCultBiotech®, #HM2358, Wayne, PA), and CD68 (macrophage marker, Cell Signaling, #97778S, Danvers, MA) to characterize relevant immune infiltrates and AAT polymer cell source6
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IgA (Immunoglobulin Alpha Heavy Chain) (B-Cell Marker) Antibody [HISA43]
$249.00 – $539.00Catalog Number:3493-MSM1Gene:IGHA1Application:Flow Cytometry, Immunofluorescence, ImmunohistochemistryReactivity:HumanHost:MouseIsotype:IgG1
Publication History:
Published in final edited form as: Eur Respir J. 2025 Jan 2;65(1):2400839. doi: 10.1183/13993003.00839-2024
