PD-L1+ Neutrophils mediate Susceptibility during Systemic Inflammatory Response in Non-Alcoholic Fatty Liver Disease
Curation statements for this article:-
Curated by eLife
eLife Assessment
This important study highlights the key role of NK cells and PD-L1+ neutrophils in worsening sepsis responses in the context of of MASH (metabolic dysfunction-associated steatohepatitis). While the data are solid, the overall evidence for the role of neutrophils in mediating this effect, which is based on a choline-deficient high-fat diet model of various knockouts or selective ablation of immune cell types, remains incomplete. The study will be of interest to researchers in immunopathological disease mechanisms.
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (eLife)
Abstract
Non-alcoholic fatty liver disease (NAFLD) is a pathological condition affecting individuals worldwide. Patients with NAFLD are more susceptible to systemic inflammation, such as sepsis, which accelerates liver damage. However, the immunological mechanisms that trigger the hyper-inflammatory responses in individuals with NAFLD remain unknown. In the present study, we reported that short-term HFCD (Choline Deficient High Fat Diet)-fed mice, which did not show significative signs of hepatic damage and inflammation in the first two weeks, are more susceptible to mortality after lipopolysaccharide (LPS) challenge. Mechanistically, endotoxemic mice show an excessive accumulation of NK-producing IFN-γ cells in liver tissue triggering the recruitment and polarization of a distinct neutrophil subset, characterized by high PD-L1 expression and massive TNFα production. Remarkably, genetic inhibition of IFN-γ or pharmacological blockade of PD-L1 effectively modulated the excessive recruitment of these neutrophils to the liver and TNFα release, thereby preventing hepatic damage and reducing the severity of host mortality. Thus, these results support the design of novel effective strategies to control hyperinflammatory responses in patients with HFCDs and consequently prevent hepatic damage and mortality..
Article activity feed
-
eLife Assessment
This important study highlights the key role of NK cells and PD-L1+ neutrophils in worsening sepsis responses in the context of of MASH (metabolic dysfunction-associated steatohepatitis). While the data are solid, the overall evidence for the role of neutrophils in mediating this effect, which is based on a choline-deficient high-fat diet model of various knockouts or selective ablation of immune cell types, remains incomplete. The study will be of interest to researchers in immunopathological disease mechanisms.
-
Reviewer #1 (Public review):
Summary:
By using an established NAFLD model, choline-deficient high-fat diet, Barros et al show that LPS challenge causes excessive IFN-γ production by hepatic NK cells which further induces recruitment and polarization of a PD-L1 positive neutrophil subset leading to massive TNFα production and increased host mortality. Genetic inhibition of IFN-γ or pharmacological blockade of PD-L1 decreases recruitment of these neutrophils and TNFα release, consequently preventing liver damage and decreasing host death.
Since NAFLD is often accompanied by chronic, low-grade inflammation, it can lead to an overactive but dysfunctional immune response and increase the body's overall susceptibility to infections, therefore this is a very important research question.
Weaknesses:
I have quite a lot of concerns with this …
Reviewer #1 (Public review):
Summary:
By using an established NAFLD model, choline-deficient high-fat diet, Barros et al show that LPS challenge causes excessive IFN-γ production by hepatic NK cells which further induces recruitment and polarization of a PD-L1 positive neutrophil subset leading to massive TNFα production and increased host mortality. Genetic inhibition of IFN-γ or pharmacological blockade of PD-L1 decreases recruitment of these neutrophils and TNFα release, consequently preventing liver damage and decreasing host death.
Since NAFLD is often accompanied by chronic, low-grade inflammation, it can lead to an overactive but dysfunctional immune response and increase the body's overall susceptibility to infections, therefore this is a very important research question.
Weaknesses:
I have quite a lot of concerns with this manuscript. One of those is that the authors did not indeed show that the seen effect is really due to NAFLD itself. The role of choline is already known in the context of sepsis since its deficiency (which can be observed in about two weeks through deterioration of liver structure and function) leads to body organ dysfunction both in humans and animals. Nolan and Vilayat in 1968 showed that the hepatic injury and mortality due to endotoxinaemic shock induced by intraperitoneal injection of LPS was significantly increased in adult female Holtzman rats fed on a choline-deficient diet. Therefore, in order to really show that the effect is mediated due to NAFLD some other diet model must be used (e.g. high-fat, high-fructose, and high-cholesterol diet).
-
Reviewer #2 (Public review):
Summary:
This is an extremely interesting mouse study, trying to understand how sepsis is tolerated during obesity/NAFLD. The researchers combine a well-established model of NASH (Choline-deficiency with High Fat Diet) with a sepsis model (IP injection of 10mg/kg LPS), leading to dramatic mortality in mice. Using this model, they characterize the complex contributions of immune cells. Specifically, they find that NK-cells and Neutrophils contribute the most to mortality in this model due to IFNG and PD-L1+ Neutrophils.
Strengths:
The biggest strength of the manuscript is how clear the primary phenotypes/endpoints of their model are. Within 6 hours of LPS injection, there is a stark elevation of liver inflammation and damage, which is exacerbated by a High Fat/CholineDeficient diet (HFCD). And after 1 day, …
Reviewer #2 (Public review):
Summary:
This is an extremely interesting mouse study, trying to understand how sepsis is tolerated during obesity/NAFLD. The researchers combine a well-established model of NASH (Choline-deficiency with High Fat Diet) with a sepsis model (IP injection of 10mg/kg LPS), leading to dramatic mortality in mice. Using this model, they characterize the complex contributions of immune cells. Specifically, they find that NK-cells and Neutrophils contribute the most to mortality in this model due to IFNG and PD-L1+ Neutrophils.
Strengths:
The biggest strength of the manuscript is how clear the primary phenotypes/endpoints of their model are. Within 6 hours of LPS injection, there is a stark elevation of liver inflammation and damage, which is exacerbated by a High Fat/CholineDeficient diet (HFCD). And after 1 day, almost all of the mice die. Using these endpoints, the authors were able to identify which cells were critical for mortality in the model and the specific mediators involved.
Weaknesses:
A few key details regarding the experimental design and interpretation are missing.
Most important is the choice of a high-fat diet with choline deficiency. I believe this model was chosen because the experiments are shorter and typically result in a liver inflammatory phenotype with not as clear of an adipose/obesity phenotype. I actually think it is typically considered a NASH (Non-alcoholic Steatohepatitis) model. I don't think the manuscript includes any data regarding the physiology of these mice that you would expect in an obesity model: body weight, liver weight, blood glucose, etc.
You should include a description in the methods for how the survival studies were conducted. Were the mice just checked on once a day for death, or were there other endpoints for euthanasia, like severe weight loss?
The measurement of IFNG and TNF in tissue throughout the manuscript seemed inconsistent. For example, IFNG in Figure 3A is 0.05pg/g for Chow+LPS, and 0.15pg.g for HFCD+LPS. But in Figure 4H, Chow+LPS is 0.18pg/g and HFCD+LPS is 0.18pg/g, so there is no effect of HFCD in the IgG controls. Also, in Figure 4I and 4J, the TNF values are dramatically different for the controls (0.1 vs 1pg/g).
You can't conclude that CD4+ and CD8+ T cells or monocytes don't play a role in liver damage from your data, because you did not measure liver damage, only mortality. I understand using mortality as an endpoint, but without ALT/AST measurements or histology, it's hard to say what exactly happened in the livers.
I'm not sure the authors can conclude that neutrophils expressing PD-L1 live longer in the hepatic environment from an in vitro experiment. I think this is an interesting result in terms of crosstalk between these two cell types, but I'm not sure that in vivo the neutrophils would live longer.
-
Reviewer #3 (Public review):
Summary:
The authors investigated how non-alcoholic fatty liver disease (NAFLD) influences liver damage during endotoxemia (a condition characterized by elevated endotoxins, like lipopolysaccharide or LPS, in the bloodstream) using a mouse model. Mice with NAFLD were given a moderate dose of LPS, which intensified liver inflammation and mortality compared to controls. The study concludes that targeting neutrophil activity and TNF-α signaling could be a promising approach to reducing excessive inflammation and liver injury in NAFLD patients experiencing endotoxemia. This can have important implications for the treatment but I think the manuscript requires revisions.
Strengths:
(1) The study presents both in vivo and ex vivo assay and results to support their hypothesis.
(2) Several cell types and their …
Reviewer #3 (Public review):
Summary:
The authors investigated how non-alcoholic fatty liver disease (NAFLD) influences liver damage during endotoxemia (a condition characterized by elevated endotoxins, like lipopolysaccharide or LPS, in the bloodstream) using a mouse model. Mice with NAFLD were given a moderate dose of LPS, which intensified liver inflammation and mortality compared to controls. The study concludes that targeting neutrophil activity and TNF-α signaling could be a promising approach to reducing excessive inflammation and liver injury in NAFLD patients experiencing endotoxemia. This can have important implications for the treatment but I think the manuscript requires revisions.
Strengths:
(1) The study presents both in vivo and ex vivo assay and results to support their hypothesis.
(2) Several cell types and their interaction with each other have been analyzed.
(3) The authors made use of the publicly available databases.
Weaknesses:
(1) Some figures contradict each other.
(2) Some of the cause-and-effect presentations need additional experiments and different approaches to be proven correct.
(3) Candidate/mechanism selection strategies are not very clear.
-
-
-
