| 1. |
- Ali, Mohamad N., et al.
(författare)
-
Osteopontin Expression in Small Airway Epithelium in Copd is Dependent on Differentiation and Confined to Subsets of Cells
- 2019
-
Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 9
-
Tidskriftsartikel (refereegranskat)abstract
- Osteopontin (OPN) plays a role in inflammation via recruitment of neutrophils and tissue remodeling. In this study, we investigated the distribution of OPN-expressing cells in the airway epithelium of normal lung tissue and that from patients with chronic obstructive pulmonary disease (COPD). OPN was detected on the epithelial cell surface of small airways and in scattered cells within the epithelial cell layer. Staining revealed higher OPN concentrations in tissue showing moderate to severe COPD compared to that in controls. In addition, OPN expression was confined to goblet and club cells, and was absent from ciliated and basal cells as detected via immunohistochemistry. However, OPN expression was up-regulated in submerged basal cells cultures exposed to cigarette smoke (CS) extract. Cell fractioning of air-liquid interface cultures revealed increased OPN production from basal compartment cells compared to that in luminal fraction cells. Furthermore, both constitutive and CS-induced expression of OPN decreased during differentiation. In contrast, cultures stimulated with interleukin (IL)-13 to promote goblet cell hyperplasia showed increased OPN production in response to CS exposure. These results indicate that the cellular composition of the airway epithelium plays an important role in OPN expression and that these levels may reflect disease endotypes in COPD.
|
|
| 2. |
- Erjefält, Jonas S., et al.
(författare)
-
Diffuse alveolar damage patterns reflect the immunological and molecular heterogeneity in fatal COVID-19
- 2022
-
Ingår i: EBioMedicine. - : Elsevier BV. - 2352-3964. ; 83
-
Tidskriftsartikel (refereegranskat)abstract
- Background: Severe COVID-19 lung disease exhibits a high degree of spatial and temporal heterogeneity, with different histological features coexisting within a single individual. It is important to capture the disease complexity to support patient management and treatment strategies. We provide spatially decoded analyses on the immunopathology of diffuse alveolar damage (DAD) patterns and factors that modulate immune and structural changes in fatal COVID-19. Methods: We spatially quantified the immune and structural cells in exudative, intermediate, and advanced DAD through multiplex immunohistochemistry in autopsy lung tissue of 18 COVID-19 patients. Cytokine profiling, viral, bacteria, and fungi detection, and transcriptome analyses were performed. Findings: Spatial DAD progression was associated with expansion of immune cells, macrophages, CD8+ T cells, fibroblasts, and (lymph)angiogenesis. Viral load correlated positively with exudative DAD and negatively with disease/hospital length. In all cases, enteric bacteria were isolated, and Candida parapsilosis in eight cases. Cytokines correlated mainly with macrophages and CD8+T cells. Pro-coagulation and acute repair were enriched pathways in exudative DAD whereas intermediate/advanced DAD had a molecular profile of elevated humoral and innate immune responses and extracellular matrix production. Interpretation: Unraveling the spatial and molecular immunopathology of COVID-19 cases exposes the responses to SARS-CoV-2-induced exudative DAD and subsequent immune-modulatory and remodeling changes in proliferative/advanced DAD that occur side-by-side together with secondary infections in the lungs. These complex features have important implications for disease management and the development of novel treatments. Funding: CNPq, Bill and Melinda Gates Foundation, HC-Convida, FAPESP, Regeneron Pharmaceuticals, and the Swedish Heart & Lung Foundation.
|
|
| 3. |
- Hvidtfeldt, Morten, et al.
(författare)
-
Airway hyperresponsiveness reflects corticosteroid-sensitive mast cell involvement across asthma phenotypes
- 2023
-
Ingår i: Journal of Allergy and Clinical Immunology. - : Elsevier BV. - 0091-6749. ; 152:1, s. 4-116
-
Tidskriftsartikel (refereegranskat)abstract
- Background: Airway hyperresponsiveness is a hallmark of asthma across asthma phenotypes. Airway hyperresponsiveness to mannitol specifically relates to mast cell infiltration of the airways, suggesting inhaled corticosteroids to be effective in reducing the response to mannitol, despite low levels of type 2 inflammation. Objective: We sought to investigate the relationship between airway hyperresponsiveness and infiltrating mast cells, and the response to inhaled corticosteroid treatment. Methods: In 50 corticosteroid-free patients with airway hyperresponsiveness to mannitol, mucosal cryobiopsies were obtained before and after 6 weeks of daily treatment with 1600 μg of budesonide. Patients were stratified according to baseline fractional exhaled nitric oxide (FENO) with a cutoff of 25 parts per billion. Results: Airway hyperresponsiveness was comparable at baseline and improved equally with treatment in both patients with FENO-high and FENO-low asthma: doubling dose, 3.98 (95% CI, 2.49-6.38; P < .001) and 3.85 (95% CI, 2.51-5.91; P < .001), respectively. However, phenotypes and distribution of mast cells differed between the 2 groups. In patients with FENO-high asthma, airway hyperresponsiveness correlated with the density of chymase-high mast cells infiltrating the epithelial layer (ρ, −0.42; P = .04), and in those with FENO-low asthma, it correlated with the density in the airway smooth muscle (ρ, −0.51; P = .02). The improvement in airway hyperresponsiveness after inhaled corticosteroid treatment correlated with a reduction in mast cells, as well as in airway thymic stromal lymphopoietin and IL-33. Conclusions: Airway hyperresponsiveness to mannitol is related to mast cell infiltration across asthma phenotypes, correlating with epithelial mast cells in patients with FENO-high asthma and with airway smooth muscle mast cells in patients with FENO-low asthma. Treatment with inhaled corticosteroids was effective in reducing airway hyperresponsiveness in both groups.
|
|
| 4. |
- Jogdand, Prajakta, et al.
(författare)
-
Eosinophils, basophils, and type 2 immune microenvironments in COPD-affected lung tissue
- 2020
-
Ingår i: European Respiratory Journal. - : European Respiratory Society (ERS). - 0903-1936 .- 1399-3003. ; 55:4
-
Tidskriftsartikel (refereegranskat)abstract
- Although elevated blood or sputum eosinophils are present in many patients with chronic obstructive pulmonary disease (COPD), uncertainties remain regarding the anatomical distribution pattern of lung-infiltrating eosinophils. Basophils have remained virtually unexplored in COPD. This study mapped tissue-infiltrating eosinophils, basophils, and eosinophil-promoting immune mechanisms in COPD-affected lungs. Surgical lung tissue and biopsies from major anatomical compartments were obtained from COPD patients with severity grades GOLD I-IV; never-smokers/smokers served as controls. Automated immunohistochemistry and in-situ hybridization identified immune cells, the type 2 immunity marker GATA3, and eotaxins (CCL11, CCL24). Eosinophils and basophils were present in all anatomical compartments of COPD-affected lungs and increased significantly in very severe COPD. The eosinophilia was strikingly patchy, and focal eosinophil-rich microenvironments were spatially linked with GATA3+ cells, including Th2 lymphocytes and type 2 innate lymphoid cells. A similarly localised and IL-33/ST2-dependent eosinophilia was demonstrated in influenza-infected mice. Both mice and patients displayed spatially confined eotaxin signatures with CCL11+ fibroblasts and CCL24+ macrophages. In addition to identifying tissue basophilia as a novel feature of advanced COPD, the identification of spatially confined eosinophil-rich type 2 microenvironments represents a novel type of heterogeneity in the immunopathology of COPD that will likely have implications for personalised treatment.
|
|
| 5. |
- Schmid, Johannes M., et al.
(författare)
-
Basophil sensitivity reflects long-term clinical outcome of subcutaneous immunotherapy in grass pollen-allergic patients
- 2021
-
Ingår i: Allergy: European Journal of Allergy and Clinical Immunology. - : Wiley. - 0105-4538. ; 76:5, s. 1528-1538
-
Tidskriftsartikel (refereegranskat)abstract
- Background: Allergic rhinoconjunctivitis is a public health problem. Allergen Immunotherapy is an effective and safe treatment, that modifies the natural course of allergic disease and induces long-term tolerance. Objective: To correlate basophil and antibody biomarkers of subcutaneous immunotherapy to clinical outcomes and cellular changes in target tissue. Methods: Adults suffering from allergic rhinoconjunctivitis due to grass pollen allergy were randomized to receive subcutaneous immunotherapy (n = 18) or to an open control group (n = 6). Patients reported daily symptom and medication scores and weekly rhinitis related quality of life scores during four pollen seasons. Biomarkers were measured every 3 months for three years treatment and every 6 months in the follow-up year. Nasal and cutaneous allergen challenge tests were performed annually. Leukocyte subsets were assessed in nasal mucosa biopsies at baseline and after treatment. Results: Subcutaneous immunotherapy led to a 447-fold decrease in basophil sensitivity during the first treatment year. This remained 100-fold lower than baseline during the 3 year-treatment period and 10-fold lower during the follow-up year (n = 18, P =.03). Decrease in basophil sensitivity after three weeks of treatment predicted long-term improvement in seasonal combined symptom and medication scores (ῥ=−0.69, P =.0027) during three years of treatment. AUC of IgE-blocking factor correlated to nasal allergen challenge (ῥ = 0.63, P =.0012) and SPT (ῥ = 0.45, P =.03). Plasma cell numbers in the nasal mucosa increased during treatment (P =.02). Conclusion: Decrease in basophil sensitivity after three weeks of subcutaneous allergen immunotherapy predicted the clinical outcome of this treatment.
|
|
| 6. |
- Siddhuraj, Premkumar, et al.
(författare)
-
Dynamically upregulated mast cell CPA3 patterns in chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis
- 2022
-
Ingår i: Frontiers in Immunology. - : Frontiers Media SA. - 1664-3224. ; 13
-
Tidskriftsartikel (refereegranskat)abstract
- Background: The mast cell-specific metalloprotease CPA3 has been given important roles in lung tissue homeostasis and disease pathogenesis. However, the dynamics and spatial distribution of mast cell CPA3 expression in lung diseases remain unknown. Methods: Using a histology-based approach for quantitative spatial decoding of mRNA and protein single cell, this study investigates the dynamics of CPA3 expression across mast cells residing in lungs from control subjects and patients with severe chronic obstructive pulmonary disease (COPD) or idiopathic lung fibrosis (IPF). Results: Mast cells in COPD lungs had an anatomically widespread increase of CPA3 mRNA (bronchioles p < 0.001, pulmonary vessels p < 0.01, and alveolar parenchyma p < 0.01) compared to controls, while granule-stored CPA3 protein was unaltered. IPF lungs had a significant upregulation of both mast cell density, CPA3 mRNA (p < 0.001) and protein (p < 0.05), in the fibrotic alveolar tissue. Spatial expression maps revealed altered mast cell mRNA/protein quotients in lung areas subjected to disease-relevant histopathological alterations. Elevated CPA3 mRNA also correlated to lung tissue eosinophils, CD3 T cells, and declined lung function. Single-cell RNA sequencing of bronchial mast cells confirmed CPA3 as a top expressed gene with potential links to both inflammatory and protective markers. Conclusion: This study shows that lung tissue mast cell populations in COPD and IPF lungs have spatially complex and markedly upregulated CPA3 expression profiles that correlate with immunopathological alterations and lung function. Given the proposed roles of CPA3 in tissue homeostasis, remodeling, and inflammation, these alterations are likely to have clinical consequences.
|
|
| 7. |
- Siddhuraj, Premkumar, et al.
(författare)
-
Histology-based blood leukocyte profiling reveals parallel Th2 and Th17 signatures in seasonal allergic rhinitis
- 2022
-
Ingår i: Acta Oto-Laryngologica. - : Informa UK Limited. - 1651-2251 .- 0001-6489. ; 142:9-12, s. 696-704
-
Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Allergic rhinitis (AR), a common condition in the westernized world, is suggested to be more immunologically complex than the archetypical 'Th2' inflammation. New approaches are needed to decode this complexity.AIMS/OBJECTIVES: In this study, we explored a novel histology-based analysis for circulating blood leukocyte profiling in 16 patients with seasonal AR outside and during the pollen season.MATERIAL AND METHODS: Leukocytes were purified with minimal ex-vivo artefacts, embedded into agarose-paraffin pellets for immunohistochemistry-based immune cell profiling. Blood leukocyte mapping was performed.RESULTS: Samples collected during the pollen season had statistically increased eosinophils, neutrophils, monocytes, and CD8+ T-lymphocytes compared to the off-season baseline. In contrast, no change was observed for CD20+ B-lymphocytes and CD3+ T-lymphocytes. Subclassification of CD4+ T-helper cells demonstrated a parallel and significant expansion of Th2 and Th17-cells during the pollen season, while Th1-cells remained unchanged. Whereas absolute basophils numbers were unaltered, the basophil markers GATA2 and CPA3 increased during the pollen season.CONCLUSIONS AND SIGNIFICANCE: This study introduces a novel and applicable method for systemic immune cell screening and provides further evidence of complex and parallel Th2 and Th17-immune signatures in seasonal AR. It also forwards GATA2 and CPA3 as potential biomarkers for ongoing allergic inflammation.
|
|
| 8. |
- Siddhuraj, Premkumar, et al.
(författare)
-
Lung Mast Cells Have a High Constitutive Expression of Carboxypeptidase A3 mRNA That Is Independent from Granule-Stored CPA3
- 2021
-
Ingår i: Cells. - : MDPI AG. - 2073-4409. ; 10:2
-
Tidskriftsartikel (refereegranskat)abstract
- The mast cell granule metalloprotease CPA3 is proposed to have important tissue homeostatic functions. However, the basal CPA3 mRNA and protein expression among mast cell populations has remained poorly investigated. Using a novel histology-based methodology that yields quantitative data on mRNA and protein expression at a single-cell level, the present study maps CPA3 mRNA and protein throughout the MCT and MCTC populations in healthy skin, gut and lung tissues. MCTC cells had both a higher frequency of CPA3 protein-containing cells and a higher protein-staining intensity than the MCT population. Among the tissues, skin MCs had highest CPA3 protein intensity. The expression pattern at the mRNA level was reversed. Lung mast cells had the highest mean CPA3 mRNA staining. Intriguingly, the large alveolar MCT population, that lack CPA3 protein, had uniquely high CPA3 mRNA intensity. A broader multi-tissue RNA analysis confirmed the uniquely high CPA3 mRNA quantities in the lung and corroborated the dissociation between chymase and CPA3 at the mRNA level. Taken together, our novel data suggest a hitherto underestimated contribution of mucosal-like MCT to baseline CPA3 mRNA production. The functional consequence of this high constitutive expression now reveals an important area for further research.
|
|
| 9. |
- Siddhuraj, Premkumar
(författare)
-
Lung Microenvironments Influence on Mast Cell Plasticity and Spatial Distribution of Immune cells
- 2021
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Inflammatory airway diseases and lower respiratory tract infections are leading causes of severe illness, disability and death worldwide. Much of today´s hypotheses about the underlying inflammatory processes and pathogenic contribution by immune cells come from in vitro experiments or animal models. As a consequence, our knowledge of the actual immunopathological events in real patient tissues is surprisingly limited. To grasp the true nature of the disease it is therefore important to assess the phenotypes and interplay of immune cells under disease-relevant in vivo conditions. The overarching aim of the present study is to investigate how lung microenvironments dictate immune cell phenotypes, plasticity and their spatial coordination in health and airway diseases. Focus is on mast cells, a key player in immune surveillance and tissue homeostasis, and cells associated with type 2 immunity. The methodological focus is on innovative computer integrated histology-based approaches that have been applied on human lung explant models, humanized mouse models (NSG-SGM3) and clinically relevant lung tissue samples from patients with COPD, CF, IPF, and COVID-19. Individuals with no history of chronic lung diseases served as control.Papers 1 and 2 profoundly alter the view on mast cell protease heterogeneity by showing that, rather than the prevailing division of mast cells into binary MCt and MCtc protease profiles, both the mRNA expression and protein granule storage of tryptase, chymase and carboxypeptidase A3 are under a fine-tuned regulation by the local tissue microenvironment. These studies also provide strong indications that individual proteases may be selectively released already at healthy base-line conditions and then further up-regulated in COPD, IPF, CF, and COVID-19. Paper 3 represents the first extensive exploration of eosinophils, basophils, and the type 2 immunity surrogate marker GATA3 across multiple anatomical sites in COPD-affected lungs. Computerized quantification and spatial analysis revealed a surprisingly patchy eosinophil distribution that co-localized with GATA3, basophil, and ILC2-containing type 2 microenvironments. Importantly, the occurrence of eosinophils, basophils and patchy type 2 microenvironments correlated with disease severity. In Paper 4 a cutting-edge humanized mouse model (NSG-SGM3) was used to show that sophisticated highly spatially organized human leukocyte tissue patterns spontaneously develop in mouse lung and lymphoid tissues. Although some of the patterns had a striking resemblance to human COPD, confounding species cross reactivity events must be taken into account before establishing humanized immune system mouse models for any human disease. In a nutshell, the present thesis reveals several new important aspects of mast cell protease plasticity, type 2 immunity, and leukocyte pattern formation and how these features are dynamically controlled by the local tissue microenvironment in health and disease. As such, the thesis also provides a powerful example of the need for “direct” explorations of patient tissues in order to understand the true complexity of inflammatory diseases and infections.
|
|
