Increased Neutrophil H 2 O 2 Production and Enhanced Pulmonary Clearance of Klebsiella pneumoniae in G6PD A- Mice

Benjamin E Zuchelkowski; Hernán F Peñaloza; Zeyu Xiong; Ling Wang; Eugenia Cifuentes-Pagano; Elizabeth Rochon; Minying Yang; Sebastien Gingras; Mark T Gladwin; Janet S Lee

doi:10.21203/rs.3.rs-3931558/v1

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Increased Neutrophil H 2 O 2 Production and Enhanced Pulmonary Clearance of Klebsiella pneumoniae in G6PD A- Mice

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Increased Neutrophil H 2 O 2 Production and Enhanced Pulmonary Clearance of Klebsiella pneumoniae in G6PD A- Mice

Benjamin E Zuchelkowski, Hernán F Peñaloza, Zeyu Xiong, Ling Wang, Eugenia Cifuentes-Pagano, Elizabeth Rochon, Minying Yang, Sebastien Gingras, Mark T Gladwin and Janet S Lee

Research square

03/11/2024

DOI: 10.21203/rs.3.rs-3931558/v1

PMCID: PMC10980108

PMID: 38559268

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https://doi.org/10.21203/rs.3.rs-3931558/v1View

Preprint (Author's original)This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

Abstract

The X-linked A- variant (rs1050828, Val68Met) in G6PDX accounts for glucose-6-phosphate (G6PD) deficiency in approximately 11% of African American males. This common, hypomorphic variant may impact pulmonary host defense and phagocyte function during pneumonia by altering levels of reactive oxygen species produced by host leukocytes. We used CRISPR-Cas9 technology to generate novel mouse strain with "humanized" G6PD A- variant containing non-synonymous Val68Met single nucleotide polymorphism. Male hemizygous or littermate wild-type (WT) controls were inoculated intratracheally with K. pneumoniae (KP2 serotype, ATCC 43816 strain,103 CFU inoculum). We examined leukocyte recruitment, organ bacterial burden, bone marrow neutrophil and macrophage (BMDM) phagocytic capacity, and hydrogen peroxide (H2O2) production. Unexpectedly, G6PD-deficient mice showed decreased lung bacterial burden (p=0.05) compared to controls 24-h post-infection. Extrapulmonary dissemination and bacteremia were significantly reduced in G6PD-deficient mice 48-h post-infection. Bronchoalveolar lavage fluid (BALF) IL-10 levels were elevated in G6PD-deficient mice (p=0.03) compared to controls at 24-h but were lower at 48-h (p=0.03). G6PD A- BMDMs show mildly decreased in vitro phagocytosis of pHrodo-labeled KP2 (p=0.03). Baseline, but not stimulated, H2O2 production by G6PD A- neutrophils was greater compared to WT neutrophils. G6PD A- variant demonstrate higher basal neutrophil H2O2 production and are protected against acute Klebsiella intrapulmonary infection.

Details

Title: Subtitle: Increased Neutrophil H 2 O 2 Production and Enhanced Pulmonary Clearance of Klebsiella pneumoniae in G6PD A- Mice
Creators: Benjamin E Zuchelkowski - University of Pittsburgh School of Medicine
Hernán F Peñaloza - University of Pittsburgh School of Medicine
Zeyu Xiong - Washington University in St. Louis
Ling Wang - University of Iowa
Eugenia Cifuentes-Pagano - Pittsburgh Heart, Lung, Blood and Vascular Medicine Institute
Elizabeth Rochon - University of Maryland, Baltimore
Minying Yang - Pittsburgh Heart, Lung, Blood and Vascular Medicine Institute
Sebastien Gingras - University of Pittsburgh School of Medicine
Mark T Gladwin - University of Maryland, Baltimore
Janet S Lee - Washington University in St. Louis School of Medicine
Resource Type: Preprint
Publication Details: Research square
DOI: 10.21203/rs.3.rs-3931558/v1
PMID: 38559268
PMCID: PMC10980108
Grant note: R01 HL168775 / NHLBI NIH HHS R01 HL125886 / NHLBI NIH HHS R01 HL098032 / NHLBI NIH HHS UH3 HL143192 / NHLBI NIH HHS
Language: English
Date posted: 03/11/2024
Academic Unit: Orthopedics and Rehabilitation
Record Identifier: 9984580301202771

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