Epithelix Sàrl

18, Chemin des Aulx – CH-1228 Plan les Ouates – Genève - Switzerland

+41 (0) 22 794 65 15 / Fax: +41 (0) 22 794 65 17

epithelix@epithelix.com – www.epithelix.com

3D Human epithelial models to study respiratory virus pathogenesis

Samuel Constant, Ph.D., COO

Respidart, 30/11/2018

Cellular Structures of Upper and Lower Airways

3D ALI airway epithelia

MucilAir™: Long shelf life in vitro Airway Tissues

Isolation, amplification & seeding

Air-Liquid interface, differentiation

Basal Cells

Goblet Cells

Ciliated Cells

Mucus

Primary Human Cells Ready-to-use

Fully differentiated

Airway Epithelium

Nose

Trachea

Bronchi

MucilAir™

Shelf-life of 1 year

MucilAir™-HF

Several versions are available:

✓ MucilAir™-HF-Normal

✓ MucilAir™-HF-Asthma

✓ MucilAir™-HF-COPD (Chronic Obstructive Pulmonary Disease)

✓ MucilAir™-HF-CF (Cystic Fibrosis)

✓ MucilAir™-HF-Allergic Rhinitis

MucilAir™-HF: Long shelf-life co-culture model

MucilAir™

Human Fibroblasts (HF)

Electron Microscopy

Photo Courtesy of Charles River Laboratories (www.criver.com)

Mucociliary clearance analysis - MucilAir

Microbeads of 30 microns are seeded onto the apical surface of MucilAir. The particle movement

is then tracked using dedicated software for calculating velocities of beads’ movement.

Mucociliary Clearance Analysis

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Mucociliary clearance analysis - MucilAir-CF

Mucociliary Clearance Analysis

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Comparison of particles velocities between MucilAir™-Normal, MucilAir™-COPD and MucilAir™-CF

The Main Characteristics of MucilAir™

▪ It has a unique shelf-life of one year

▪ It mimics the morphology and functions of the normal

human airway epithelium.✓ Active Ion transport

✓ Metabolic activity

▪ Epithelia from several different pathologies are available

(Asthma, COPD, Cystic Fibrosis, Allergic Rhinitis, etc.)

▪ It is easy to handle and maintain

▪ Serum free medium

▪ Worldwide Shipping

SmallAir™ : Long shelf life in vitro Airway Tissues

Isolation, amplification & seeding

Air-Liquid interface, differentiation

Primary Human Cells Ready-to-use

Fully differentiated

Airway Epithelium

Small Airways SmallAir™

SmallAir™ - Histology - Immunohistochemistry

MucilAir™ SmallAir™

A)

B)

C)

MucilAir™ (Bronchial) SmallAir™ (Small-Airways)

H/E-Alcian Blue

Muc-5Ac(Goblet Cells marker)

CC-10(Club Cells marker)

Huang et al. Establishment and Characterization of an in vitro Human Small Airway Model (SmallAir™). Eur. J. Pharm. Biopharm. 2017

In vitro Antiviral Testing using MucilAir™/SmallAir™

HRV-A 16/41/49/55

HRV-B 14/48

HRV-C 2/6/7/8/11/12/1315/18/23/28/29/47

Rhinovirus

Influenza A (H1N1 and H3N2)

Influenza

EV-68

OC 43

Coronavirus

RSV-ARSV-B

Respiratory Syncytial Virus

Others

Metapneumovirus

Respiratory Enterovirus

Efficient replication of respiratory viruses

Essaidi-Laziosi et al. Propagation of respiratory viruses in human airway epithelia reveals persistent virus-specific signatures JACI. 2017

Faster replication of RSVB in MucilAir™ vs SmallAir™

M u c i lA i rT M

- P o o l R S V B d i lu t i o n

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T i m e p o s t - i n o c u l a t i o n ( h o u r s )

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Respiratory viruses have different cell tropisms

▪ All viruses tested found in ciliated cells

▪ H3N2 found also in goblet and basal cells

Ciliated cells Viruses Cell nuclei

Dynamic Exposure Systems :

Static Exposure Systems :

Gas or smoke

Solids (Tablets)Liquids - Solutions

Nebulizing chambers

Antiviral testing: Exposure systems

Inhibition of virus replication using antivirals

RV-A16 EV-D68

H1N1 H3N2

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Boda et al. Antiviral drug screening by assessing epithelial functions and innate immune responses in human 3D airway epithelium modelAntiviral research. 2018

Antivirals prevent disruption of barrier function

Virus-induced impairement of mucociliary function

All other virusesRV-B, OC43 and non infected

All viruses block mucociliary clearance except rhinovirus B and coronavirus OC43

Antivirals prevent impairment of Mucociliary function

Conclusion

▪ ALI 3D Human Airway Models like MucilAir™ and SmallAir™ are

useful tools to study viral pathogenesis

✓ Isolation and characterization of difficult to growth viruses

✓ Identification of cytotoxic viruses (H3N2 and EV-D68)

✓ Visualization of viral persistence in absence of immune cells

▪ Novel test strategies using ALI 3D Human Airway Models allows

evaluation of broad spectrum of antivirals

Thanks for your attention

▪ Paul Alouani

▪ Sacha Benaoudia

▪ Dr. Bernadett Boda

▪ Rosy Bonfante

▪ Emilie Ferreira

▪ Ina Fureraj

▪ Matia Gojun

▪ Carole Bertinetti

▪ Mireille CaulFuty

▪ Ophélie Verbeke

▪ Laurent Wiszniewski

▪ Dr. Ludovic Wiszniewski

Swiss LabFrench Lab

▪ Dr. Song Huang

▪ Xiao-Yann Huang

▪ Faten Hussein

▪ Marc Lanzillo

▪ Melany Monachino

▪ Rebecca Pimenta

▪ Jimmy Vernaz

▪ Karin Weber

▪ Prof. Caroline Tapparel

▪ Dr. Manel Essaidi-Laziosi