Related Experiment Video

Updated: May 3, 2026

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

Published on: April 5, 2017

Primary application of PPE68 of Mycobacterium tuberculosis

Zhiling Dong¹, Lei Xu¹, Jing Yang¹

¹Department of Pathogenic Organisms, Chongqing Medical University, Chongqing, China; Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, China.

Human Immunology

|February 18, 2014

Abstract:

PPE68 protein is absent from BCG and the attenuated strains of Mycobacterium tuberculosis (MTB). In this study, the shuttle plasmid pBudCE4.1/PPE68/OriM was constructed and transformed into BCG to obtain PPE68 recombination BCG (PPE68-rBCG), and BALB/c mice were immunized with PPE68-rBCG to evaluate the immunological characterization of PPE68-rBCG. The level of lgG2a, IFN-γ, IL-12 and IL-4 in serum of immunized mice were detected, the proliferation response of spleen lymphocyte were measured, the frequency of CD4(+), CD8(+) and CD4(+)/CD8(+) were determined, and the spleen and lung tissue were prepared for pathological analysis. PPE68-rBCG was constructed successfully and could induce powerful Th1 immune response in mice. Besides, we took the purified recombination PPE68 (rPPE68) protein as diagnostic antigen to detect pulmonary tuberculosis patients (n=252) and extrapulmonary tuberculosis patients (n=66). We also used anti-PPE68 polyclonal antibody as coating antibody to detect specific antigen in the same serum samples. Our data provide an experimental basis for potential application of rPPE68 in the diagnosis of tuberculosis, especially for extrapulmonary tuberculosis.

Insights

Recombinant PPE68 BCG (PPE68-rBCG) successfully induced a strong Th1 immune response in mice. The study also shows potential for using recombinant PPE68 protein in diagnosing tuberculosis, particularly extrapulmonary tuberculosis.

Area of Science:

Immunology
Microbiology
Biotechnology

Background:

PPE68 protein is absent in BCG and attenuated Mycobacterium tuberculosis (MTB) strains.
Recombinant BCG expressing PPE68 (PPE68-rBCG) was developed to study its immunological properties.
The study aimed to evaluate the immune response induced by PPE68-rBCG and the diagnostic potential of rPPE68 protein.

Purpose of the Study:

To construct and characterize PPE68 recombination BCG (PPE68-rBCG).
To evaluate the immunological responses induced by PPE68-rBCG in BALB/c mice.
To assess the diagnostic utility of recombinant PPE68 (rPPE68) protein for tuberculosis detection.

Main Methods:

Construction of shuttle plasmid pBudCE4.1/PPE68/OriM and transformation into BCG.
Immunization of BALB/c mice with PPE68-rBCG and analysis of serum cytokines (lgG2a, IFN-γ, IL-12, IL-4).
Assessment of spleen lymphocyte proliferation, T-cell populations (CD4+, CD8+), and pathological analysis of spleen and lung tissues. Diagnostic evaluation using rPPE68 antigen and anti-PPE68 antibody in patient serum samples.

Main Results:

Successful construction of PPE68-rBCG, which induced a robust Th1 immune response in mice.
Demonstrated significant levels of IgG2a, IFN-γ, and IL-12, indicative of a Th1 response.
rPPE68 protein showed potential as a diagnostic antigen for pulmonary and extrapulmonary tuberculosis detection.

Conclusions:

PPE68-rBCG is a promising candidate for inducing a potent Th1 immune response.
Recombinant PPE68 protein holds potential for the diagnosis of tuberculosis, especially extrapulmonary forms.
The findings provide an experimental basis for the clinical application of rPPE68 in tuberculosis diagnostics.

Read full analysis

More Related Videos

An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis

An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis

Published on: August 16, 2021

Protective Efficacy and Pulmonary Immune Response Following Subcutaneous and Intranasal BCG Administration in Mice

Protective Efficacy and Pulmonary Immune Response Following Subcutaneous and Intranasal BCG Administration in Mice

Published on: September 19, 2016

Related Concept Videos

Pulmonary Tuberculosis V

Pulmonary Tuberculosis V

Medical management of tuberculosis (TB) patients involves a comprehensive approach that includes diagnosis, treatment, and monitoring. The specific strategies can vary depending on the type of tuberculosis (latent or active), the patient's overall health status, and other considerations.
Latent tuberculosis infection occurs when TB bacteria are present in a person's body, but are not causing illness or symptoms. It is not contagious, and preventive treatment is crucial to avoid the...

Pulmonary Tuberculosis IV

Pulmonary Tuberculosis IV

Tuberculosis, more commonly referred to as TB, is an infectious disease stemming from Mycobacterium tuberculosis. While it primarily impacts the lungs, TB can also affect other body areas. Given its severity and global impact, timely and accurate diagnosis is crucial for controlling its spread and improving patient outcomes.
Several diagnostic approaches are used to detect TB. The conventional method is the Tuberculin Skin Test (TST), also known as the Mantoux test. However, this method has...

Pulmonary Tuberculosis III

Pulmonary Tuberculosis III

Tuberculosis (TB) is a contagious infection primarily affecting the lung parenchyma but which can also affect other body parts. TB can be classified based on disease development, presentation, and the affected anatomical site.
The first classification is based on the development of the disease, and it includes the following categories:

Pulmonary Tuberculosis I

Pulmonary Tuberculosis I

Tuberculosis, often called TB, is a contagious illness primarily caused by Mycobacterium tuberculosis. It mainly affects the lung parenchyma but can also impact other body parts.
Causative Organism
The primary infectious agent causing tuberculosis is Mycobacterium tuberculosis, a slow-growing, acid-fast, aerobic rod that exhibits sensitivity to heat and ultraviolet light. Instances of Mycobacterium bovis and Mycobacterium avium contributing to the development of TB infection are rare.
Mode of...

Pulmonary Tuberculosis II

Pulmonary Tuberculosis II

Tuberculosis, or TB, is a bacterial infectious disease caused by Mycobacterium tuberculosis. While its primary impact is on the lungs, leading to pulmonary tuberculosis, it can also affect various other organs, a condition referred to as extrapulmonary tuberculosis.
Here is a detailed explanation of its pathophysiology:
Transmission: The process begins when a person inhales droplet nuclei containing M. tuberculosis. These are typically released into the air when an individual with pulmonary or...

Personal Protective Equipment

Personal Protective Equipment

Personal protective equipment (PPE) is unique clothing or equipment worn by an employee to minimize or prevent exposure to infectious agents. PPE creates a barrier between the employee and the infectious materials. PPE must be readily available in the patient care area. PPE includes gloves, gowns and aprons, masks and respirators, goggles, face shields, shoes, and headcovers:

Articles linked to this work by shared authors, journal, and citation graph.

Patient-Specific MRI-Based Right Ventricle Models Using Different Zero-Load Diastole and Systole Geometries for Better Cardiac Stress and Strain Calculations and Pulmonary Valve Replacement Surgical Outcome Predictions.

PloS one·2016

Crossover phenomena of percolation transition in evolution networks with hybrid attachment.

Chaos (Woodbury, N.Y.)·2016

Relation Between C-X-C Motif Chemokine Receptor 4 Levels and the Presence and Extent of Angiographic Coronary Collaterals in Patients With Chronic Total Coronary Occlusion.

The American journal of cardiology·2016

Quantify patient-specific coronary material property and its impact on stress/strain calculations using in vivo IVUS data and 3D FSI models: a pilot study.

Biomechanics and modeling in mechanobiology·2016

Analysis of Genomewide DNA Methylation Reveals Differences in DNA Methylation Levels between Dormant and Naturally as well as Artificially Potentiated Pedicle Periosteum of Sika Deer (Cervus nippon).

Journal of experimental zoology. Part B, Molecular and developmental evolution·2016

Value of corneal epithelial and Bowman's layer vertical thickness profiles generated by UHR-OCT for sub-clinical keratoconus diagnosis.

Scientific reports·2016

See all related articles