Adenosine metabolism inhibits airway inflammation and remodeling in asthma model induced by Aspergillus fumigatus

doi:10.21203/rs.3.rs-2448772/v1

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Adenosine metabolism inhibits airway inflammation and remodeling in asthma model induced by Aspergillus fumigatus

https://doi.org/10.21203/rs.3.rs-2448772/v1

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published 13 Jul, 2023

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Background: Given that severe inflammatory reaction is an important manifestation of asthma, it would be helpful to understand the pathological mechanism of asthma to determine the endogenous anti-inflammatory events of asthma. Recent studies have found that energy metabolites in the body are related to some inflammatory diseases, but little is known about their role in asthma.

Method: In this study, an asthma model was established by intratracheal instillation of Aspergillus fumigatus extract in Ecto-5'-Nucleotidase (CD73) gene–knockout and wild-type mice. Multiple analyses from bronchoalveolar lavage fluid (BALF) were used to determine the levels of cytokines and chemokines. Immunohistochemistry was used to detect subcutaneous fibrosis and inflammatory cell infiltration. Finally, adenosine 5’-(α, β-methylene) diphosphate (APCP), a CD73 inhibitor, was pumped subcutaneously before Aspergillus attack to observe the infiltration of inflammatory cells and subcutaneous fibrosis to clarify its therapeutic effect.

Result: PAS staining showed that CD73 knockout inhibited pulmonary epithelial cell proliferation and bronchial fibrosis induced by Aspergillus extract. The genetic destruction of CD73 significantly reduced the production of Th2 cytokines, interleukin (IL)-4, IL-6, IL-13, chemokine (C-C motif) ligand 5 (CCL5), eosinophil chemokine, neutrophil IL-17, and granulocyte colony-stimulating factor (G-CSF). In addition, exogenous adenosine supplementation increased airway inflammation. Finally, the CD73 inhibitor APCP was administered to reduce inflammation and subcutaneous fibrosis.

Conclusion: Elevated adenosine metabolism plays an inflammatory role in asthma, and CD73 could be a potential therapeutic target for asthma.

adenosine

asthma

CD73

metabolism

Asthma is a chronic inflammatory disease of airways, the main clinical signs of which include wheezing, breathlessness, cough, and chest tightness. Inhaled corticosteroids, leukotriene modifiers, long-acting β2mimetics, theophylline, and anticholinergic drugs are commonly used to relieve the symptoms of asthma. Inhaled corticosteroids (ICS) are still the first‑line treatment for chronic asthma. However, long-term high-dose inhalation of corticosteroids may lead to adverse reactions [1]. Therefore, it is important to elucidate the mechanism of asthma development to determine effective therapeutic targets for asthma.

Asthma is characterized by increased eosinophils, IgE production, the proliferation of airway smooth muscle cells, and mucus overproduction by goblet cells. In the microenvironment of asthma airway inflammation, proinflammatory cytokines such as IL-4, IL-5, IL-13, chemokines such as CXCL1, CCL3, CCL4, MIP-1α, MCP-1 [2–5], and other signals such as adenosine monophosphate [6, 7], leukotrienes (LT), and prostaglandin D₂ (PGD₂) [8, 9] participate in the acute attack of asthma and the regulation of airway inflammation. Mediators such as prostaglandin E₂ (PGE₂) and adenosine play an anti-inflammatory role in airway inflammation [10]. The loss of local pro-/anti-inflammatory balance in the airway leads to airway inflammation in asthma.

Adenosine level is significantly increased in patients with asthma [11, 12]. Metabolic pathways, such as acetate, adenosine, alanine, hippurate, succinate, threonine, and trans-aconitate, are involved in hypoxia, oxidative stress, and inflammation in asthma [13]. Plasma taurine, bile acid, nicotinamide, and adenosine 5' -monophosphate are involved in the activation and immune pathways of asthma inflammation [14]. Endogenous adenosine is produced in inflammatory environments. ecto-5'-nucleotidase (CD73) is a protein located on the plasma membrane, which catalyzes the conversion of extracellular nucleotides to membrane-permeable nucleosides and hydrolyzes AMP to adenosine. It has been suggested that CD73 influences the level of adenosine and ATP to affect airway inflammation and fibrosis. In this study, we examined the role of adenosine and its metabolizing enzyme, CD73, in Aspergillus fumigatus-extract-induced asthma. We found that inhibition of CD73 could relieve airway inflammation and cytokine production, thereby providing a new treatment possibility for asthma patients with elevated adenosine.

Animals

C57BL/6, CD73^+/+, and CD73^−/− mice were bred at the Experimental Animal Center of the Beijing Institute of Heart, Lung, and Blood Vessel Diseases. All animal experiments were approved by the animal ethics committee of the Experimental Animal Center of the Beijing Institute of Heart, Lung, and Blood Vessel Diseases.

Asthma model

Female mice (6–8 weeks old) were sensitized by intratracheal instillations of 5 µL A.f. extract (Hollister-Stier Laboratories, Spokane, WA) in a total volume of 20 µL under isoflurane anesthesia every other day for 3 weeks. Littermates that only received saline were the control group [15, 16].

In another set of experiments, mice received subcutaneously implanted adenosine 5’-(α, β-methylene) diphosphate (APCP, CD73 inhibitor, 40 mg/kg) through Alzet pumps (2004, delivery rate: 0.25 µL/h for 4 weeks) under isoflurane anesthesia. Implantation was performed 12 h before the A.f. challenge procedure. Wild-type littermate mice that only received saline were used as the control group. After the APCP pump procedure, adenosine was intratracheally instilled one hour after the A.f. challenge.

Bronchoalveolar lavage fluid and lung histopathology

After the final challenge, the total BALF was collected. The supernatant was stored at − 80°C for subsequent cytokine quantification, and the remaining cells were used for Wright-Giemsa staining. Neutrophils, macrophages, eosinophils, and lymphocytes were quantified via oil microscopy at a magnification of 1,000×, and the number of each cell type was determined after counting a total of 400 cells per high-power field. Lungs were fully infiltrated with 10% formalin for paraffin embedding, and 5-µm-thick sections were stained with hematoxylin and eosin (H&E) and Alcian Blue Periodic acid Schiff (AB-PAS) staining or Masson’s trichrome staining. Epithelium height was determined using the NIS-ELEMENTS quantitative automatic program (Nikon, Japan) as previously described. Morphological evaluation of inflammatory infiltrates was based on myeloperoxidase (MPO; 1:200 dilution, Abcam) as a marker of peribronchial neutrophils, α-smooth muscle actin (SMA; 1:200 dilution, Abcam) as a marker of myofibroblast activation, and transforming growth factor (TGF)-β1 (1:200 dilution, Abcam) as a marker of lung fibrosis, and was performed at light microscopy at a magnification of 400×. Analysis of the percentage of positive cells was determined as the ratio of the positive staining area to the total section area by use of the NIS-ELEMENTS quantitative automatic program (Nikon, Japan).

Measurement of cytokines and chemokines

The supernatants in BALF and cell culture were collected to detect cytokines and chemokines using a Luminex multiplex cytokine assay (Bio-Rad Laboratories, Inc, Berkeley, CA) in line with the manufacturer’s instructions. The data were analyzed using Bio-Plex Manager software (Bio-Rad 200 System). Serum IgE was measured using a commercial enzyme-linked immunosorbent assay (ELISA; R&D Systems) following the manufacturer’s instructions.

RNA extraction and real-time PCR

Total RNA was obtained from the right lung lobe using TRIzol reagent (Invitrogen, Carlsbad, CA) in accordance with the manufacturer’s directions. A total of 2 µg RNA was reverse-transcribed into cDNA with M-MLV and random primers (Promega, Madison, WI). Specific oligonucleotide primers were added to the buffer along with 2 µL of reverse-transcribed cDNA sample. The cDNA was amplified using the following cycling parameters: the mixture was first incubated for 4 min at 94°C, then cycled 40 times at 94°C for 30 s and 60°C for 30 s, and elongated at 72°C for 30 s. An iQ5 system (Bio-Rad Laboratories, Inc, Berkeley, CA) with SYBR Green I (Takara, Shiga, Japan) was used for real-time quantitative PCR analysis. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the internal control. The level of mRNA was normalized to GAPDH expression, and the results were analyzed by the 2^−ΔΔCt method (supplementary table 1).

Determination of bronchial hyperresponsiveness

The mice were anesthetized, and their airway resistance to different doses of methacholine (0–100 mg/mL) was measured using an invasive plethysmograph (Buxco, London, U.K.). Airway resistance was recorded at 10-s intervals for 5 min, and the average values were represented as cm⸱H₂O/mL/s.

Neutrophil cell culture

Neutrophils were isolated from bone marrow using RPMI 1640 medium supplemented with 10% FBS. Adenosine and ATP ratios co-cultured with neutrophils in the presence or absence of an A2A receptor antagonist (ZM241385, Sigma) was measured by high-performance liquid chromatography (HPLC) and mass spectrum as described previously.

Statistical analysis

Data are presented as mean ± standard error of the mean (SEM). Statistical differences between the two groups were assessed by one-way analysis of variance (ANOVA) followed by Scheffe’s post-hoc test for selected pairs. For all statistical analyses, P < 0.05 was considered statistically significant. GraphPad Prism 5 (GraphPad Software Inc., San Diego, CA) was used.

CD73 deficiency suppresses peribronchial fibrosis

Chronic airway inflammation causes pathological lung fibrosis. To determine the role of CD73 deficiency in lung fibrosis induced by the A.f. challenge, Masson’s trichrome and immunohistochemical staining were used to determine the extent of peribronchial fibrosis. The lung fibrotic area was reduced in CD73-deficiency mice compared with wild-type mice (Fig. 1A). Myofibroblast activation in the lungs of CD73-deficiency mice was also reduced (Fig. 1B). Furthermore, the level of profibrotic mediator TGF-β1 in the lungs of CD73-knockout mice was reduced compared with that in wild-type mice (Fig. 1C). These results indicate that CD73 deficiency suppresses peribronchial fibrosis.

CD73 deficiency suppresses airway inflammation

To investigate whether CD73 deficiency has a role in A.f.-induced airway inflammation, we used CD73-knockout mice to evaluate airway immune response. H&E staining and Giemsa staining were performed to determine immune cell infiltration. Inflammatory cell infiltration in lungs and BALF were suppressed in CD73-deficiency mice compared with wild-type mice (Figs. 2A&2B). In accordance with these results, IgE and Th2 cytokines were reduced in CD73-deficiency mice compared with wild-type mice (Figs. 2C&2D).

Goblet cell hyperplasia and mucus production were evaluated by PAS staining (Fig. 2E). The epithelium height of CD73-deficiency mice was lower than that of wild-type mice (Fig. 2F). Mucus production was further examined by the mRNA of the mucus glycoproteins Muc1 and Muc5AC. CD73-deficiency mice showed a reduction in mucus production compared with wild-type mice (Fig. 2G). The adenosine to ATP ratio was reduced in CD73-deficiency airway inflammation (Fig. 2H). These results demonstrate that CD73 deficiency suppresses airway inflammation.

Reduced number of neutrophils in CD73-deficiency mice

Neutrophils play an important role in innate immune defense. As the first line of circulating leukocytes, neutrophils migrate toward damaged/infected tissue and lead to increased inflammatory response. Excessive activation of neutrophils results in chronic inflammation. Therefore, we examined whether neutrophils were suppressed in CD73 deficiency. Neutrophil infiltration in BALF and lungs was examined by Giemsa staining and MPO immunohistochemistry staining (Figs. 3A&3C). CD73-deficiency mice showed a significant reduction in the number of neutrophils compared with wild-type mice (Fig. 3B). To further demonstrate whether CD73 deficiency limits the production of neutrophils in airway inflammation, key mediators that regulate neutrophils production, such as granulocyte colony-stimulating factor (G-CSF) and IL-17, were evaluated. G-CSF and IL-17 levels were significantly reduced in CD73-deficiency mice compared with wild-type mice (Fig. 3D). These data suggest that CD73 deficiency reduces the influx of neutrophils to BALF and lungs.

Adoptive transfer of A.f.-treated CD73-deficiency neutrophils limits inflammatory mediators

To investigate whether the decreased airway inflammation was mediated by neutrophils, neutrophils from CD73-deficiency mice or wild-type mice treated with A.f. were adoptively transferred to wild-type mice subsequently challenged with A.f. to induce airway inflammation. Neutrophils deficient in CD73 limited inflammation both in the lungs and BALF (Figs. 4A&4B). Apart from the reduction in Th2 cytokines (Fig. 4C), chemokine levels were reduced in CD73-deficiency neutrophils compared with wild-type neutrophils (Fig. 4D). These data demonstrate that neutrophils from CD73-deficiency mice downregulate airway inflammation.

Abnormal adenosine to ATP ratio impairs neutrophil function

It has been reported that A2aR activation suppresses neutrophil function [17], while CD73 is a key enzyme in the hydrolysis of ATP to adenosine. The expression of A2aR mRNA was evaluated in CD73-knockout mice and wild-type mice after the A.f. challenge. As shown in Fig. 5A, A2aR mRNA was increased in CD73-knockout mice compared with wild-type mice. These data suggest that A2aR is involved in the neutrophil-related reduction of airway inflammation in CD73-deficiency mice. Thus, we hypothesized that a low ratio of adenosine to ATP promotes A2aR activation in neutrophils. Expression of A2aR and production of chemokines by neutrophils were evaluated on different adenosine to ATP ratios with or without ZM241385 (ZM, an adenosine A2A receptor antagonist).

As shown in Fig. 5B, the expression of A2aR was enhanced with a decreasing ratio of adenosine to ATP. In addition, the adenosine A2A receptor antagonist ZM inhibited A2aR mRNA expression. Next, we evaluated the production of chemokines by neutrophils. As shown in Fig. 5C, the ZM group demonstrated inhibited chemokine secretion compared with the adenosine and ATP groups. It can be suggested that blocking adenosine A2A receptor shows a suppressive effect on chemokines production. These data suggest that lower adenosine to ATP ratio impairs neutrophil chemotaxis through activated adenosine A2A receptors.

CD73 reduces airway inflammation

To further confirm whether the therapeutic application of a CD73 inhibitor suppresses chronic airway inflammation, APCP, a specific inhibitor of CD73, was subcutaneously implanted prior to the Aspergillus extract challenge (Fig. 6A). Peribronchial and perivascular inflammation as well as goblet cell hyperplasia were inhibited in APCP-treated mice (Fig. 6B). Immune cell infiltration in BALF was reduced in APCP-treated mice (Fig. 6C). In accordance with these results, the Th2 cytokines in BALF were reduced in APCP-treated mice (Fig. 6D), accompanied by a decrease in GATA3 mRNA expression in the lungs (Fig. 6F). There was a decreased airway resistance to methacholine in APCP-treated mice compared with the control mice (Fig. 6E).

To evaluate whether exogenous adenosine supplementation may abolish APCP-dependent reduced airway inflammation, adenosine was administered after APCP implantation. We found that adenosine increased airway inflammation and airway resistance. In addition, GATA3 increased after adenosine supplementation (Fig. 6F). Overall, these data indicate a therapeutic role of CD73 inhibitor in chronic airway inflammation.

The present study demonstrated the protective role of CD73 deficiency in a model of asthma induced by Aspergillus fumigatus. Our data showed the suppression of airway inflammation when CD73 was inhibited or deficient, suggesting that inhibition of CD73 could be a potential therapy for asthma.

Recent reports have indicated that allergens such as A. fumigatus may lead to asthma [18, 19]. Increased asthma symptoms in children and adults are indeed associated with increased levels of Aspergillus [20]. Intratracheal delivery of A. fumigatus conidia into the airways of C57BL/6 mice elicits neutrophilic asthma, with strong Th17 response, eosinophil-related inflammation, and significant IgE production, but very mild airway hyperresponsiveness [21]. In addition, the characteristics of asthma vary according to the genetic background of mice. Namely, inflammatory cells and cytokines are increased in C57BL/6 mice, while airway hyperresponsiveness is increased in BALB/c mice [22]. Compared with saline-treated mice, we found that the serum IgE, BALF neutrophils, and mucus production of C57BL/6 wild-type mice treated with Aspergillus extract increased significantly. It has been reported that A.f.-induced asthma inflammation is characterized by Th2 cytokine production, goblet cell hyperplasia, and airway hyperreactivity in C57BL/6 mice [23]. The expression of cytokines such as IL-6 is increased in A.f.-induced airway inflammation [24]. Our data confirmed that A.f. treatment upregulated the expression of IL-6. These data indicate that an A.f.-induced Th2 airway inflammation model has been established.

It has been reported that chronic inflammation involves all types of airway cells, including increased mucus secretion by goblet cells, increased production of cytokines and chemokines, and increased thickness of the airway muscle wall [25]. Recent studies have shown that adenosine plays an important role in the development and progression of airway inflammation. The effect of adenosine on inflammation in asthma depends on adenosine receptors [26]. Adenosine activates adenosine receptors on immune cells, affects their function, and plays an important role in asthma. CD73 is a transmembrane protein that plays an important pathophysiological role in conversion of AMP to adenosine. Recent studies have demonstrated that CD73 has an important role in pulmonary disease depend on types of injury [27]. It is unclear whether CD73 deficiency plays an important role in airway inflammation induced by A.f. We used CD73-knockout mice to characterize the role of CD73 in the A.f.-induced airway inflammatory process. We showed that A.f.-challenged CD73-knockout mice displayed a phenotype of reduced immune cell infiltration and Th2 immune response accompanied by decreased mucus gland metaplasia. A.f. treatment of CD73-knockout mice led to the reduced production of the Th2 cytokines, IL-4, IL-6, and IL-13. It can be suggested that CD73 deficiency protects from A.f.-induced airway inflammation response. These findings suggest that CD73 deficiency plays a protective role against A.f.-induced airway inflammation.

Recent reports have shown that neutrophils play an important role in killing pathogens and removing cellular debris, which causes inflammation [28]. Excessive activation of neutrophils leads to prolonged inflammation and aggravation of asthma [29]. Neutrophil elastase limits allergic airway inflammation and hyperresponsiveness [30]. However, it is unclear whether CD73 in neutrophils plays an important role in inflammation. Therefore, we utilized the adoptive transfer of CD73-deficient neutrophils to wild-type mice and challenged them with A.f. extract. We found that CD73 in neutrophils was able to reduce inflammatory cell infiltration and Th2 cytokines production. It has been reported that the ability of neutrophils to kill pathogens is enhanced in lung infection [31]. However, chemotactic activity and activation of neutrophils are crucial for inflammation response. Our data demonstrate that the lack of CD73 in neutrophils treated with A.f. reduces chemokines secretion, which is related to the migration of immune cells to the damaged sites. This indicates that CD73 in neutrophils mediates the increase of inflammation through chemotaxis upregulation. In summary, the lack of CD73 in neutrophils protects from excessive activation of neutrophils to reduce airway inflammation.

We found that the expression of the A2A receptor was increased during the A.f. challenge of CD73-deficiency mice. In addition, we found that neutrophils treated with different ratios of adenosine and ATP upregulated the expression of the A2A receptor in response to Aspergillus. It has been suggested that abnormal adenosine metabolism activates the A2A receptor. In A2AR knockout mice induced by LPS, inflammation increases with the increase of neutrophil infiltration into BALF [32]. To prove whether the activation of the A2A receptor affects neutrophil chemotaxis, we incubated neutrophils with A2A receptor antagonist ZM241385 under different adenosine to ATP ratios. The data showed that the blockage of the A2A receptor suppressed the expression of chemokines in vitro. Therefore, abnormal adenosine metabolism activates the A2A receptor on neutrophils, thereby leading to the release of mediators and cytokines, which induces airway inflammation.

It is well-known that chronic airway inflammation develops into fibrosis after prolonged exposure to allergy. The link between CD73 and tissue fibrosis has widely been studied in many diseases, but the mechanism is still unclear. For example, in transverse aortic constriction–induced heart failure, CD73 plays anti-inflammatory and antifibrotic effects by activating the adenosine A2a receptor [33]. After unilateral ischemia–reperfusion injury or folic acid treatment, perivascular cell CD73 inhibits the transformation of perirenal interstitial myofibroblasts, inhibits inflammation, and prevents progressive fibrosis [34]. However, in a CCL₄-induced hepatic fibrosis model, the mRNA expression levels of Collα1, Col3α1, and TGF-β1 in wild-type mice were much higher than those in CD73 knockout mice. Inhibiting the production of adenosine or blocking adenosine receptors may help to prevent hepatic fibrosis [35]. In bleomycin-challenged dermal fibrosis, adenosine promotes dermal fibrosis through adenosine receptor activation, and CD73 inhibitors can treat dermal fibrosis diseases, such as scleroderma [36]. The role of CD73 in pulmonary fibrosis needs further study. The bleomycin-induced pulmonary fibrosis is a widely used model, and CD73 knockout mice show increased inflammation and fibrosis of the lung. Intranasal instillations of exogenous nucleotidase decrease inflammation and fibrosis [37]. In a model of radiation-induced lung injury, pulmonary fibrosis decreased after CD73-antibody treatment [38]. Our study showed that peribrochial fibrosis was reduced in CD73 knockout mice after being challenged with Aspergillus extract. Compared with wild-type mice, the activation of myofibroblasts and TGF-β1 in the lungs was reduced in CD7-deficiency mice. These results indicate that CD73 promotes peribronchial fibrosis.

Furthermore, to confirm the therapeutic usefulness of blocking CD73 in asthma, we used APCP, a CD73 inhibitor, to treat mice with A.f. challenge. We found that not only immune cell infiltration and mucus overproduction but also AHR was suppressed in APCP-treated mice with A.f. challenge. In addition, exogenous supplementation of adenosine aggravated airway inflammation and AHR.

In conclusion, CD73 deficiency exerts a protective effect against excessive neutrophil infiltration through the upregulated expression of the A2A receptor in Aspergillus extract–induced asthma. These results suggest that inhibition of CD73 could be a potential novel therapy for asthma.

AB-PAS, Alcian Blue Periodic acid Schiff; A.f., Aspergillus fumigatus; APCP, adenosine 5’-(α, β-methylene) diphosphate; BALF, bronchoalveolar lavage fluid; CCL5, chemokine (C-C motif) ligand 5; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; G-CSF, granulocyte colony-stimulating factor; H&E, hematoxylin and eosin; PGD2, prostaglandin D2, PGE2, prostaglandin E2

Ethics declarations

The study is reported in accordance with ARRIVE guidelines. All the methods were carried out in accordance with relevant guidelines and regulations. Animal experiments were ethically approved by the Experimental Animal Welfare and Ethics Committee of Beijing Anzhen Hospital Affiliated to the Capital Medical University. This study was performed according to the to the National Institutes of Health Guide for Care and Use of Laboratory Animals following the principial of the 3Rs (Refinement, Reduction and replacement), pain minimization and approved euthanasia protocols.

Consent for publication

Not applicable

Authorship contribution statement

T.L. designed the work, performed experiments and wrote the original manuscript; Y.L. performed mouse experiments and collected data; Z.Z. preformed methodology and collected data; L.J. performed review & editing; J.Z. preformed methodology; Z.C. performed review & editing; H.S. performed review & editing; C.P. supervised the study and substantively revised the paper. J.D. supervised the study. All authors read and approved the final manuscript.

Acknowledgments

We thank Dr. Yang Liu and Professor Aijuan Qu (Department of Pathophysiology, Capital Medical University) for help with airway resistance test; Dr. Fan Meng and Professor Zhihua Gao (Zhejiang University Interdisciplinary Institute of Neuroscience and Technology) for CD73 knockout mice; Professor Zhang jing for advice with APCP in Alzert pump. We thank LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Declaration of Competing Interest

The authors declare that there is no conflict of interest regarding the publication of this paper.

Funding

This work was supported by a Grant from the National Natural Science Foundation of China (82170409).

Availability of Data and Materials

The datasets used and/or analysed during the current study available from the corresponding author on reasonable request.

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No competing interests reported.

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published 13 Jul, 2023

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Editorial decision: Major revision
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Adenosine metabolism inhibits airway inflammation and remodeling in asthma model induced by Aspergillus fumigatus

Status:

Journal Publication

Version 1

Abstract

Figures

Background

Methods

Animals

Asthma model

Bronchoalveolar lavage fluid and lung histopathology

Measurement of cytokines and chemokines

RNA extraction and real-time PCR

Determination of bronchial hyperresponsiveness

Neutrophil cell culture

Statistical analysis

Results

CD73 deficiency suppresses peribronchial fibrosis

CD73 deficiency suppresses airway inflammation

Reduced number of neutrophils in CD73-deficiency mice

Adoptive transfer of A.f.-treated CD73-deficiency neutrophils limits inflammatory mediators

Abnormal adenosine to ATP ratio impairs neutrophil function

CD73 reduces airway inflammation

Discussion

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

Status:

Journal Publication

Version 1