The Role of a Mycobacterium Growth Inhibition Assay to Quantify Host Immune Control of M. Tuberculosis (MGIA)

The Role of a Mycobacterium Growth Inhibition Assay to Quantify Host Immune Control of M. Tuberculosis From Healthy Blood Donors and Patients With Latent or Active Tuberculosis.

The goal of this observational study is to learn about how the blood leucocytes from patients with and without exposure or disease from tuberculosis (TB) are able to kill live mycobacteria (M. tuberculosis). The main question it aims to answer is:

Is intracellular growth suppression of M. tuberculosis in peripheral blood mononuclear cells variable among healthy blood donors and higher than in patients with latent TB compared to active TB?

Study Overview

• Status: Recruiting
• Conditions: Tuberculosis; Immunologic Activity Alteration
• Intervention / Treatment: Other: No intervention.

Detailed Description

Background: The host immune response is crucial in determining the outcome after exposure to Mtb. A substantial proportion of exposed individuals (40-70%) never develop infection indicating an efficient cell mediated immune response against Mtb and early clearance by the host immune defense (Verall et al, Immunology 2014). Thus, the outcome of exposure is highly dependent on host immunity. In this study, our aim was to investigate whether the previously developed mycobacterium groth inhibition assay (MGIA) method (Andersson et al, Tuberculosis 2020) could be used to quantify the host immune response to Mtb.

Study design: In this study we will include healthy donors (n=80), patient with active TB (n=40), subjects recently exposed to active TB (n=80) and patients with latent TB (LTBI) (n=80). Exclusion criteria for all groups are known HIV infection or other immunosuppression from treatment or disease. Patients and healthy blood donors will be recruited at the departments of infectious diseases in Region Kalmar and Region Östergötland. From patients giving oral and written consent, 40 ml of blood will be collected for MGIA analysis.

Primary aim: To determine the difference in MGIA between healthy blood donors and patients with latent and active TB as well as the variation in host control of Mtb within each group.

Method: The Mycobacterial growth inhibition assay (MGIA) using human PBMCs isolated by density gradient by Lymhoprep (Axis-Shield) and SepMate tubes (StemCell Technologies) will be performed as described in study I with minor modifications. The PBMCs are infected with GFP expressing M. tuberculosis H37Rv and intracellular growth of Mtb and viability of PBMCs are assessed by live-cell imaging (Incucyte©) and measured as relative fluorescence units (RFU) during 5 days with and without stimulation with the purified protein derivate (PPD) is included. The cytokine response following exposure to gamma-irradiated H37Rv will be quantified by Olink proteomics.

• Study Type: Observational
• Enrollment (Estimated): 280

Contacts and Locations

• Study Contact: Name: Thomas Schoen, Professor; Phone Number: +48013580000; Email: tschon@hotmail.com

Study Locations

• Sweden
  • Kalmar, Sweden, 391 85
    • Recruiting
    • Kalmar County Hospital
    • Contact: Thomas Schön, Professor; Phone Number: +46103850000; Email: tschon@hotmail.com
  • Linköping, Sweden, 581 85
    • Recruiting
    • Dept of Infectious Diseases
    • Contact: Thomas Schön, Professor; Phone Number: +461010000000; Email: tschon@hotmail.com
    • Sub-Investigator: Jakob Paues, MD PhD
    • Sub-Investigator: Katarina Niward, MD PhD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Probability Sample

Study Population

Subjects who are attending the infectious disease clinics at region Östergötland och region Kalmar, Sweden.

Description

Inclusion Criteria: Healthy blood donors or subjects who will get vaccinated.

Exclusion Criteria:Known HIV infection, not Swedish citizen, chronic liver och kidney disease,immunosuppression from treatment or disease.

Study Plan

How is the study designed?

Number of groups / cohorts

4

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Healthy Individuals; Patients with a negative IGRA (Quantiferon) who are apparently healthy.	Other: No intervention.; Blood sampling for analysis of mycobacterial growth inhibition assay (MGIA) analysis.
Close contacts to patients with active TB; Close contacts to patients with active, smear positive pulmonary TB.	Other: No intervention.; Blood sampling for analysis of mycobacterial growth inhibition assay (MGIA) analysis.
Patients with (latent) tuberculosis infection; Patients without active TB who have a positive IGRA (Quantiferon Plus) result.	Other: No intervention.; Blood sampling for analysis of mycobacterial growth inhibition assay (MGIA) analysis.
Patients with active TB; Culture or PCR-verified active TB who are initiated on treatment.	Other: No intervention.; Blood sampling for analysis of mycobacterial growth inhibition assay (MGIA) analysis.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Mycobacterial inhibition of host macrophages	Mycobacterial inhibition of host macrophages	Up to 10 days incubation
Mycobacterial inhibition of host peripheral blood mononuclear cells (PBMCs)	Mycobacterial inhibition of host peripheral blood mononuclear cells (PBMCs)	Up to 10 days incubation

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Cytokine expression of host peripheral blood mononuclear cells (PBMCs)	Proteome analysis following exposure of PBMCs to gamma-irradiated M. tuberculosis	24 hours

Collaborators and Investigators

• Sponsor: Linkoeping University
• Investigators: Principal Investigator: Thomas Schoen, Professor, Linkoeping University

Study record dates

Study Major Dates

• Study Start (Actual): August 1, 2020
• Primary Completion (Estimated): March 1, 2025
• Study Completion (Estimated): December 1, 2025

Study Registration Dates

• First Submitted: August 28, 2024
• First Submitted That Met QC Criteria: August 28, 2024
• First Posted (Actual): August 30, 2024

Study Record Updates

• Last Update Posted (Actual): August 30, 2024
• Last Update Submitted That Met QC Criteria: August 28, 2024
• Last Verified: August 1, 2024

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Infections; Bacterial Infections; Bacterial Infections and Mycoses; Gram-Positive Bacterial Infections; Actinomycetales Infections; Mycobacterium Infections; Tuberculosis
• Other Study ID Numbers: 2022-06858-02

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No