Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice

Pioli, KimAnh Trang; Pioli, Peter D.

doi:10.1016/j.xpro.2023.102308

Cited by 3 publications

(2 citation statements)

References 3 publications

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“…To distinguish between circulating neutrophils in the lung microvasculature versus neutrophils that have extravasated into the lung tissue interstitium, we injected CD45.2 i.v. to label all neutrophils contained within the vascular space immediately prior to lung harvest ( 42 ). Although circulating CD45.2 pos Ly6G + neutrophil numbers were similar between WT and Stat1 −/− mice ( Fig.…”

Section: Resultsmentioning

confidence: 99%

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STAT1 Employs Myeloid Cell–Extrinsic Mechanisms to Regulate the Neutrophil Response and Provide Protection against Invasive Klebsiella pneumoniae Lung Infection

Gonzalez-Ferrer,

Peñaloza,

van der Geest

et al. 2024

ImmunoHorizons

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Klebsiella pneumoniae (KP) is an extracellular Gram-negative bacterium that causes infections in the lower respiratory and urinary tracts and the bloodstream. STAT1 is a master transcription factor that acts to maintain T cell quiescence under homeostatic conditions. Although STAT1 helps defend against systemic spread of acute KP intrapulmonary infection, whether STAT1 regulation of T cell homeostasis impacts pulmonary host defense during acute bacterial infection and injury is less clear. Using a clinical KP respiratory isolate and a pneumonia mouse model, we found that STAT1 deficiency led to an early neutrophil-dominant transcriptional profile and neutrophil recruitment in the lung preceding widespread bacterial dissemination and lung injury development. Yet, myeloid cell STAT1 was dispensable for control of KP proliferation and dissemination, because myeloid cell–specific STAT1-deficient (LysMCre/WT;Stat1fl/fl) mice showed bacterial burden in the lung, liver, and kidney similar to that of their wild-type littermates. Surprisingly, IL-17–producing CD4+ T cells infiltrated Stat1−/− murine lungs early during KP infection. The increase in Th17 cells in the lung was not due to preexisting immunity against KP and was consistent with circulating rather than tissue-resident CD4+ T cells. However, blocking global IL-17 signaling with anti–IL-17RC administration led to increased proliferation and dissemination of KP, suggesting that IL-17 provided by other innate immune cells is essential in defense against KP. Contrastingly, depletion of CD4+ T cells reduced Stat1−/− murine lung bacterial burden, indicating that early CD4+ T cell activation in the setting of global STAT1 deficiency is pathogenic. Altogether, our findings suggest that STAT1 employs myeloid cell–extrinsic mechanisms to regulate neutrophil responses and provides protection against invasive KP by restricting nonspecific CD4+ T cell activation and immunopathology in the lung.

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Section: Resultsmentioning

confidence: 99%

“…delivery 3 min prior to lungs being harvested at 24 h postinfection ( 41 ). Following CD45.2 administration, we determined tissue-resident versus circulating cells based on a method that was published previously ( 42 ). CD45.2 pos cells were considered i.v.…”

Section: Methodsmentioning

confidence: 99%

STAT1 Employs Myeloid Cell–Extrinsic Mechanisms to Regulate the Neutrophil Response and Provide Protection against Invasive Klebsiella pneumoniae Lung Infection

Gonzalez-Ferrer,

Peñaloza,

van der Geest

et al. 2024

ImmunoHorizons

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Purification and analysis of kidney-infiltrating leukocytes in a mouse model of lupus nephritis

Amo,

Kole,

Scott

et al. 2024

Methods in Cell Biology

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Comparison of lateral tail vein and retro-orbital venous sinus as routes of inoculation to study Group B streptococcal systemic infection

da Conceição Mendonça,

Lumsdaine,

Burcham

2024

Microbiol Spectr

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Murine models are commonly used to understand pathogen and host determinants of systemic infection. While these models have proven beneficial for uncovering bacterial mechanisms required for progression to invasive disease, it can be challenging to draw comparisons across studies as several different routes of infection are standardly used for these experiments. In this study, one of the leading bacterial meningeal pathogens, Streptococcus agalactiae , or Group B Streptococcus (GBS), was used to compare experimental outcomes of two commonly used routes of hematogenous infection, lateral tail vein injection and retro-orbital venous sinus injection. Here we demonstrate that both routes of infection result in systemic disease and the onset of clinical symptoms of infection. We show that retro-orbital venous sinus injection results in an initial increase in bacterial dissemination to the spleen and brain tissue of GBS-infected mice, while an increased bacterial burden was only detected in brain tissues at a later time point. Despite differences in initial dissemination and brain bacterial burden, we found that the route of infection did not significantly impact bacterial burden in the blood, kidney, spleen, heart, and lung tissues at experimental endpoints; and similarly did not impact animal health scores during infection; cytokine and proinflammatory protein abundance in the brain tissue; or overall animal survival. In summary, these findings suggest that both tail vein injection and retro-orbital venous sinus injection are viable models to study Group B streptococcal systemic infection and result in largely similar disease outcomes within our tested parameters. IMPORTANCE Streptococcus agalactiae or Group B Streptococcus (GBS) is the leading cause of invasive disease in neonates and immunocompromised adults and is implicated in severe cases of sepsis, pneumonia, and meningitis. Established murine models of hematogenous systemic infection allow for better understanding and investigation of bacterial dispersion, pathogenesis, meningeal inflammation, and interaction with the host. Here we compared two routes of infection, intravenous lateral tail vein and retro-orbital venous sinus, demonstrating that similar experimental outcomes can be observed, regardless of the route of infection for GBS, specifically. These findings help to reinforce the utility of different systemic infection models and provide insight into comparisons across different established models and how these models can be applied in microbial research.

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Retro-orbital CD45 antibody labeling to evaluate antibody-secreting cell trafficking in mice

Cited by 3 publications

References 3 publications

STAT1 Employs Myeloid Cell–Extrinsic Mechanisms to Regulate the Neutrophil Response and Provide Protection against Invasive Klebsiella pneumoniae Lung Infection

STAT1 Employs Myeloid Cell–Extrinsic Mechanisms to Regulate the Neutrophil Response and Provide Protection against Invasive Klebsiella pneumoniae Lung Infection

Purification and analysis of kidney-infiltrating leukocytes in a mouse model of lupus nephritis

Comparison of lateral tail vein and retro-orbital venous sinus as routes of inoculation to study Group B streptococcal systemic infection

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