Tropism and Pathogenesis of Influenza Virus and Coronavirus in Human Brain Explant Culture

NCT05375006 | Unknown DMC

• 1 other identifier: 2022.120
• Study Type: observational
• Target: 80
• Locations: 1 country
• Sites: 1

Brief Summary

Background: Influenza and coronavirus have been repeatedly causing pandemic recently. Like the Influenza A/H7N9 virus has caused five epidemics in China since its first detection in East China in 2013. In 2017, the previously low pathogenic avian influenza (LPAI) H7N9 virus underwent mutation in its haemagglutinin to give to a highly pathogenic avian influenza (HPAI) virus causing 32 human cases and potentially poses a threat to animal and human health. More recently, the SARS-CoV-2 pandemic has been heavily affecting the world. Therefore an effective risk assessment platform is urgently required for better pandemic preparation. Hypothesis: The tissue tropism and pathogenesis of a newly emerged infectious viruses, like the highlypathogenic influenza, like H7N9 and coronavirus, like SARS-CoV-2 would be different from that of their low pathogenic subtype and it would infect and replicate the human respiratory system more efficiently. Because of its resistance to oseltamivir for influenza and no effective antiviral for coronavirus, investigators therefore propose to set up an novel and effective risk assessment platform for emerging infectious viruses. Experimental Design: The tissue tropism and viral replication kinetics of a HPAI and LP influenza and coronavirus will be determined in ex vivo cultures of human brain and compared with their LP subtype. The replication competence and innate immune responses of influenza and coronavirus will be studied and compared with other LP virus in in vitro cultures of human brain cells and human microvascular endothelial cells (HMVEC) both isolated from human brain tissues. Expected outcomes: HPAI influenza and coronavirus particularly SARS-CoV-2 will infect and replicate the human brain tissues and cells more efficiently than their LP subtype. Besides, HPAI influenza and SARS-CoV-2 will induce dysregulated host innate immune response than the LP subtype.

Conditions

Influenza Virus; Coronavirus

Outcome Measures

Primary Outcomes (4)

• Replication kinetics of influenza virus using ex vivo cultures of human brain

  Tissue fragments will be infected with 106 TCID50/mL for 1h at 37°C and then washed with 3 ml of warm PBS for three times to remove unbound virus. To determine productive viral replication from the infected biopsy specimens, supernatants of the infected cultures will be collected at 1, 24, 48 and 72hpi and virus titers will be determined by TCID50 assay. Explant cultures will be fixed in 10% formalin at 24 and 72hpi and immunohistochemistry staining (IHC) against HB65 antibody (European Veterinary Laboratories), SARS-CoV or SARS-CoV-2 viral antibody will be performed for the detection of virus-infected cells.

  baseline

• replication efficiency and innate immune responses of influenza virus

  Cells will be infected with viruses at either MOI of 0.01 or 2 for 1h. After 1h virus adsorption, viruses will be aspirated and cells will be washed with PBS for three times, then medium will be replenished. For cells infected at MOI of 0.01, cell culture supernatant will be harvested at 1, 24, 48 and 72hpi and viral titers will be determined by TCID50 assay.

  baseline

• Replication kinetics of coronavirus using ex vivo cultures of human brain

  Tissue fragments will be infected with 106 TCID50/mL for 1h at 37°C and then washed with 3 ml of warm PBS for three times to remove unbound virus. To determine productive viral replication from the infected biopsy specimens, supernatants of the infected cultures will be collected at 1, 24, 48 and 72hpi and virus titers will be determined by TCID50 assay. Explant cultures will be fixed in 10% formalin at 24 and 72hpi and immunohistochemistry staining (IHC) against HB65 antibody (European Veterinary Laboratories), SARS-CoV or SARS-CoV-2 viral antibody will be performed for the detection of virus-infected cells.

  Baseline

• replication efficiency and innate immune responses coronavirus

  Cells will be infected with viruses at either MOI of 0.01 or 2 for 1h. After 1h virus adsorption, viruses will be aspirated and cells will be washed with PBS for three times, then medium will be replenished. For cells infected at MOI of 0.01, cell culture supernatant will be harvested at 1, 24, 48 and 72hpi and viral titers will be determined by TCID50 assay.

  Baseline

Eligibility Criteria

• Age: 1 Year - 70 Years
• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)
• Sampling Method: Non-Probability Sample

Study Population

This study will recruit participants who will undergoing elective or emergency craniotomies for intrinsic brain lesions at Prince of Wales Hospital

You may qualify if:

• Age: \> 1 year old and \< 70 years old
• Undergo elective or emergency craniotomies for intrinsic brain lesions

You may not qualify if:

• a. Samples containing infected material

Sponsors & Collaborators

• Chinese University of Hong Kong: lead
• The University of Hong Kong: collaborator

Study Sites (1)

Prince of Wales Hospital

Shatin, NT, 00000, Hong Kong

RECRUITING

Biospecimen

Retention: SAMPLES WITH DNA

Brain Tissue

MeSH Terms

Conditions

Coronavirus Infections

Condition Hierarchy (Ancestors)

Coronaviridae Infections; Nidovirales Infections; RNA Virus Infections; Virus Diseases; Infections

Study Officials

• Owen Ho Ko, PhD

  Chinese University of Hong Kong

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Owen Ho Ko, Ph.D

CONTACT

+852 26352160; ho.ko@cuhk.edu.hk

Study Design

• Study Type: observational
• Observational Model: CASE ONLY
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Assistant Professor

Study Record Dates

• First Submitted: May 12, 2022
• First Posted: May 16, 2022
• Study Start: May 26, 2022
• Primary Completion: March 31, 2024
• Study Completion: September 30, 2024
• Last Updated: August 17, 2022

Record last verified: 2022-08

Data Sharing

• IPD Sharing: Will not share

Locations

HK (1)