Characterization of the Dynamic Behavior of Neutrophils Following Influenza Vaccination

Abstract

Neutrophils are amongst the first cells to respond to inflammation and infection. Although they play a key role in limiting the dissemination of pathogens, the study of their dynamic behavior in immune organs remains elusive. In this work, we characterized in vivo the dynamic behavior of neutrophils in the mouse popliteal lymph node (PLN) after influenza vaccination with UV-inactivated virus. To achieve this, we used an image-based systems biology approach to detect the motility patterns of neutrophils and to associate them to distinct actions. We described a prominent and rapid recruitment of neutrophils to the PLN following vaccination, which was dependent on the secretion of the chemokine CXCL1 and the alarmin molecule IL-1α. In addition, we observed that the initial recruitment occurred mainly via high endothelial venules located in the paracortical and interfollicular regions of the PLN. The analysis of the spatial-temporal patterns of neutrophil migration demonstrated that, in the initial stage, the majority of neutrophils displayed a patrolling behavior, followed by the formation of swarms in the subcapsular sinus of the PLN, which were associated with macrophages in this compartment. Finally, we observed using multiple imaging techniques, that neutrophils phagocytize and transport influenza virus particles. These processes might have important implications in the capacity of these cells to present viral antigens.

The Secretive Life of Neutrophils Revealed by Intravital Microscopy

“…Intravital microscopy showed the entry of neutrophils into the popliteal lymph node (PLN) via multiple hotspots on HEV following influenza vaccination (Pizzagalli et al, 2019). Moreover, neutrophil positive for influenza virus were tracked entering into the PLN after vaccination and showed over a time of 2 h changes in their dynamic motility with a decrease in instantaneous and mean speed, directionality, displacement, and an increase in the arrest coefficient suggesting an increase in cell-to-cell interactions (Pizzagalli et al, 2019). Five distinct neutrophil migratory behaviors have been observed: flowing, arrested, patrolling, directed migration, and swarming (Pizzagalli et al, 2019).…”

“…Moreover, neutrophil positive for influenza virus were tracked entering into the PLN after vaccination and showed over a time of 2 h changes in their dynamic motility with a decrease in instantaneous and mean speed, directionality, displacement, and an increase in the arrest coefficient suggesting an increase in cell-to-cell interactions (Pizzagalli et al, 2019). Five distinct neutrophil migratory behaviors have been observed: flowing, arrested, patrolling, directed migration, and swarming (Pizzagalli et al, 2019). During swarming, neutrophils were observed forming clusters that enlarged over time in areas rich in tissue resident macrophages (Pizzagalli et al, 2019).…”

“…Five distinct neutrophil migratory behaviors have been observed: flowing, arrested, patrolling, directed migration, and swarming (Pizzagalli et al, 2019). During swarming, neutrophils were observed forming clusters that enlarged over time in areas rich in tissue resident macrophages (Pizzagalli et al, 2019). In another study using a model of skin infection, neutrophils were observed migrating to the PLN recruited by tissue resident macrophages in an IL-1β-dependent manner to control the spread of pathogens (Kastenmuller et al, 2012).…”

Neutrophil subtypes shape HIV-specific CD8 T-cell responses after vaccinia virus infection

“…3d, e ). To understand if these different neutrophil subsets have distinct biological functions, we analyzed their migratory patterns based on their directionality and arrest coefficient 25 . Indeed, we found that Nβ neutrophils had distinct migratory pattern compared to Nα subset between 4 and 5 h p.i.…”

“…To understand better the different dynamics of recruitment of both neutrophil subsets from the primary site of infection to secondary lymphoid organs, we analyzed the expression of the surface markers involved in neutrophil migration and adhesion, such as IL-1 receptor (IL-1R), CXCR2, and very-late-antigen (VLA-4) α4β1. IL-1R has a clear role in neutrophil recruitment to the peritoneal cavity 26 and migration to secondary lymphoid organs 25 . CXCR2 regulates neutrophil release from bone marrow 27 and neutrophil migration in tumor microenvironment and during severe sepsis 28 , 29 ; it has been shown that failure of CXCR2 downregulation controls neutrophil mobilization during inflammation in the peritoneum 30 .…”