scholarly journals Novel Human Lung Tissue Model for the Study of SARS-CoV-2 Entry, Inflammation and New Therapeutics

2021 ◽  
Author(s):  
Judith Grau-Expósito ◽  
David Perea ◽  
Marina Suppi ◽  
Núria Massana ◽  
Ander Vergara ◽  
...  
Keyword(s):  
Lung Tissue ◽  
Viral Entry ◽  
Human Lung ◽  
Inflammatory Responses ◽  
Alveolar Type ◽  
List Type ◽  
Human Lung Tissue ◽  
Main Cell ◽  
Anti Inflammatory ◽  
Conventional Systems

AbstractThe development of physiological models that reproduce SARS-CoV-2 infection in primary human cells will be instrumental to identify host-pathogen interactions and potential therapeutics. Here, using cell suspensions from primary human lung tissues (HLT), we have developed a platform for the identification of viral targets and the expression of viral entry factors, as well as for the screening of viral entry inhibitors and anti-inflammatory compounds. We show that the HLT model preserves its main cell populations, maintains the expression of proteins required for SARS-CoV-2 infection, and identifies alveolar type II (AT-II) cells as the most susceptible cell targets for SARS-CoV-2 in the human lung. Antiviral testing of 39 drug candidates revealed a highly reproducible system, and provided the identification of new compounds missed by conventional systems such as VeroE6. Using this model, we also show that interferons do not modulate ACE2 expression, and that stimulation of local inflammatory responses can be modulated by different compounds with antiviral activity. Overall, we present a novel and relevant physiological model for the study of SARS-CoV-2.SynopsisEx vivo physiological systems for the study of SARS-CoV-2-host interactions are scarce. Here, we establish a novel model using primary human lung tissue (HLT) for the analysis of cell tropism and identification of therapeutics.The HLT model preserves main cell subpopulations, including alveolar type-2 cells, and expression of SARS-CoV-2 entry factors ACE2, CD147, and TMPRSS2.The HLT model is readily susceptible to SARS-CoV-2 entry.Antiviral testing in the HLT model allows the identification of new candidates missed by conventional systems.Local inflammation is supported in the HLT model and offers the identification of relevant anti-inflammatory compounds for SARS-CoV-2 infection.

scholarly journals Evaluation of SARS-CoV-2 entry, inflammation and new therapeutics in human lung tissue cells

2022 ◽  
Vol 18 (1) ◽  
pp. e1010171
Author(s):  
Judith Grau-Expósito ◽  
David Perea ◽  
Marina Suppi ◽  
Núria Massana ◽  
Ander Vergara ◽  
...  
Keyword(s):  
Viral Entry ◽  
Human Lung ◽  
Inflammatory Responses ◽  
Alveolar Type ◽  
Human Lung Tissue ◽  
Physiological Models ◽  
Structural Cell ◽  
Drug Candidates ◽  
Entry Inhibitors

The development of physiological models that reproduce SARS-CoV-2 infection in primary human cells will be instrumental to identify host-pathogen interactions and potential therapeutics. Here, using cell suspensions directly from primary human lung tissues (HLT), we have developed a rapid platform for the identification of viral targets and the expression of viral entry factors, as well as for the screening of viral entry inhibitors and anti-inflammatory compounds. The direct use of HLT cells, without long-term cell culture and in vitro differentiation approaches, preserves main immune and structural cell populations, including the most susceptible cell targets for SARS-CoV-2; alveolar type II (AT-II) cells, while maintaining the expression of proteins involved in viral infection, such as ACE2, TMPRSS2, CD147 and AXL. Further, antiviral testing of 39 drug candidates reveals a highly reproducible method, suitable for different SARS-CoV-2 variants, and provides the identification of new compounds missed by conventional systems, such as VeroE6. Using this method, we also show that interferons do not modulate ACE2 expression, and that stimulation of local inflammatory responses can be modulated by different compounds with antiviral activity. Overall, we present a relevant and rapid method for the study of SARS-CoV-2.

scholarly journals IL-33 Mediates Inflammatory Responses in Human Lung Tissue Cells

2010 ◽  
Vol 185 (10) ◽  
pp. 5743-5750 ◽  
Author(s):  
Akiko Yagami ◽  
Kanami Orihara ◽  
Hideaki Morita ◽  
Kyoko Futamura ◽  
Noriko Hashimoto ◽  
...  
Keyword(s):  
Lung Tissue ◽  
Human Lung ◽  
Inflammatory Responses ◽  
Human Lung Tissue ◽  
Tissue Cells

A modified method for isolating viable alveolar type II cells from human lung tissue

1990 ◽  
Vol 132 (1) ◽  
pp. 145-146 ◽  
Author(s):  
Frans J. Van Overveld ◽  
Wilfried A. De Backer ◽  
Paul A. Vermeire
Keyword(s):  
Lung Tissue ◽  
Human Lung ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Human Lung Tissue ◽  
Alveolar Type Ii Cells ◽  
Modified Method

IL-36 induces cytokine IL-6 and chemokine CXCL8 expression in human lung tissue cells: Implications for pulmonary inflammatory responses

Cytokine ◽  
2017 ◽  
Vol 99 ◽  
pp. 114-123 ◽  
Author(s):  
Juan Zhang ◽  
Yibing Yin ◽  
Xue Lin ◽  
Xingxing Yan ◽  
Yun Xia ◽  
...  
Keyword(s):  
Lung Tissue ◽  
Human Lung ◽  
Inflammatory Responses ◽  
Human Lung Tissue ◽  
Tissue Cells

scholarly journals Cryobanking of human distal lung epithelial cells for preservation of their phenotypic and functional characteristics.

2021 ◽  
Author(s):  
Bindu Konda ◽  
Apoorva Mulay ◽  
Changfu Yao ◽  
Edo Israely ◽  
Stephen Beil ◽  
...  
Keyword(s):  
Single Cell ◽  
Lung Tissue ◽  
Human Lung ◽  
Alveolar Type ◽  
Basal Cells ◽  
Human Lung Tissue ◽  
Downstream Analysis ◽  
Long Term Preservation

The epithelium lining airspaces of the human lung is maintained by regional stem cells including basal cells of pseudostratified airways and alveolar type 2 pneumocytes (AT2) of the alveolar gas-exchange region. Despite effective methods for long-term preservation of airway basal cells, methods for efficient preservation of functional epithelial cell types of the distal gas-exchange region are lacking. Here we detail a method for cryobanking of epithelial cells from either mouse or human lung tissue for preservation of their phenotypic and functional characteristics. Flow cytometric profiling, epithelial organoid-forming efficiency, and single cell transcriptomic analysis, were used to compare cells recovered from cryopreserved tissue with those of freshly dissociated tissue. Alveolar type 2 cells within single cell suspensions of enzymatically digested cryobanked distal lung tissue retained expression of the pan-epithelial marker CD326 and the AT2 cell surface antigen recognized by monoclonal antibody HTII-280, allowing antibody-mediated enrichment and downstream analysis. Isolated AT2 cells from cryobanked tissue were comparable with those of freshly dissociated tissue both in their single cell transcriptome and their capacity for in vitro organoid formation in 3D cultures. We conclude that the cryobanking method described herein allows long-term preservation of distal human lung tissue for downstream analysis of lung cell function and molecular phenotype, and is ideally suited for creation of an easily accessible tissue resource for the research community.

scholarly journals Lung Immune Tone Regulation by the Gut-Lung Immune Axis: Short-chain Fatty Acid Receptors FFAR2 and FFAR3, and IL-1β Expression Profiling in Mouse and Human Lung

2020 ◽  
Author(s):  
Qing Liu ◽  
Xiaoli Tian ◽  
Daisuke Maruyama ◽  
Mehrdad Arjomandi ◽  
Arun Prakash
Keyword(s):  
Lung Tissue ◽  
Gut Microbiome ◽  
Human Lung ◽  
Expression Patterns ◽  
Metabolic Programming ◽  
Short Chain ◽  
Alveolar Type ◽  
Human Lung Tissue

ABSTRACTMicrobial metabolites produced by the gut microbiome, such as short-chain fatty acids (SCFA), can influence both local intestinal and distant lung physiology and response to injury. However, how lung immune activity is regulated by SCFAs is unknown. We examined fresh human lung tissue and observed the presence of SCFAs with large inter-individual and even intra-lobe variability. In vitro, SCFAs were capable of modifying the metabolic programming in both resting and LPS-exposed alveolar macrophages (AM). Additionally, since we hypothesized that lung immune tone could be defined through priming of the inflammasome (aka signal 1), we interrogated naïve mouse lungs for pro-IL-1β message and localized its presence within the alveolar space in situ, specifically in AM subsets, and in close proximity to alveolar type 2 epithelial (AT2) cells. We established that metabolically active gut microbiota, that produce SCFAs, can transmit LPS and SCFAs to the lung (potential sources of signal 1), and thereby could regulate lung immune tone and metabolic programming. To understand how murine lung cells sensed and upregulated IL-1β in response to gut-microbiome factors, we determined that in vitro, AM and AT2 cells expressed SCFA receptors, FFAR2, FFAR3, and IL-1β but with different expression patterns and LPS-inducibility. Finally, we observed that IL-1β, FFAR2 and FFAR3 were expressed both in isolated human AM and AT2 cells ex-vivo, but in fresh human lung sections in situ, only AM expressed IL-1β at rest and after LPS challenge. Together, this translational study using mouse and human lung tissue and cells supports an important role for the gut microbiome and SCFAs in regulating lung immune tone.

Mast Cell Heterogeneity in Human Lung Tissue

1989 ◽  
Vol 77 (3) ◽  
pp. 297-304 ◽  
Author(s):  
F. J. Van Overveld ◽  
L. A. M. J. Houben ◽  
F. E. M. Schmitz du Moulin ◽  
P. L. B. Bruijnzeel ◽  
J. A. M. Raaijmakers ◽  
...  
Keyword(s):  
Mast Cell ◽  
Mast Cells ◽  
Alcian Blue ◽  
Lung Tissue ◽  
Human Lung ◽  
Immunoglobulin E ◽  
Prostaglandin D2 ◽  
Leukotriene C4 ◽  
Human Lung Tissue ◽  
No Release

1. In this study mast cells were found to comprise 2.1% of total cells recovered by enzymatic digestion of human lung tissue. 2. This mast cell population consisted of 79% formalin-sensitive, Alcian Blue-positive mast cells and 21% formalin-insensitive, Alcian Blue-positive mast cells. 3. By the use of centrifugal elutriation and subsequent Percoll gradient centrifugation, separate mixed cell populations could be obtained in which the mast cell constituents were either of the formalin-sensitive or -insensitive type. 4. Cell suspensions in which formalin-sensitive cells comprised 97% of mast cells contained approximately 1.34 pg of histamine per mast cell, whereas in preparations in which mast cells were 84% formalin-resistant the histamine content was approximately 4.17 pg of histamine per mast cell. 5. The histamine release upon anti-immunoglobulin E challenge of formalin-sensitive mast cells was greater than the release by formalin-insensitive mast cells. 6. After challenge with opsonized zymosan, only formalin-sensitive mast cells were able to release histamine. 7. Leukotriene C4 release was observed when formalin-sensitive mast cells were challenged with antiimmunoglobulin E. Formalin-insensitive mast cells showed no release of leukotriene C4. 8. Prostaglandin D2 release was observed when formalin-insensitive mast cells were challenged with antiimmunoglobulin E. Formalin-sensitive mast cells showed no release of prostaglandin D2.

The Glucocorticosteroid, Budesonide, Partially Blocks Histamine Release from Human Lung Tissue in Vitro

Allergy ◽  
1986 ◽  
Vol 41 (5) ◽  
pp. 319-326 ◽  
Author(s):  
H. Bergstrand ◽  
B. Lundquist ◽  
B.-Å. Petersson
Keyword(s):  
Histamine Release ◽  
Lung Tissue ◽  
Human Lung ◽  
Human Lung Tissue
Sign in / Sign up

Export Citation Format

Share Document