Related Experiment Video
Updated: Aug 8, 2026

Separation and Fractionation of Culture Filtrate Proteins (CFPs) from Mycobacterium tuberculosis
Published on: July 11, 2025
Characterization of mycobacterial antigens and antibodies in circulating immune complexes from pulmonary tuberculosis
A Raja1, P R Narayanan, R Mathew
1Department of Immunology, Tuberculosis Research Centre, Madras, India.
Insights
Circulating immune complexes (CICs) show elevated levels in tuberculosis patients. Measuring antibodies against antigens under 70 kDa within CICs can help diagnose pulmonary tuberculosis, even in bacteriologically negative cases.
Area of Science:
- Immunology
- Infectious Diseases
- Biochemistry
Background:
- Circulating immune complexes (CICs) play a role in various diseases.
- Pulmonary tuberculosis diagnosis can be challenging, especially in bacteriologically negative cases.
- Understanding CIC composition and antibody profiles is crucial for diagnostic markers.
Purpose of the Study:
- To investigate the levels and diagnostic utility of CICs in pulmonary tuberculosis.
- To compare different methods for CIC measurement.
- To characterize antibodies within CICs for potential diagnostic markers.
Main Methods:
- Measurement of CICs using C1q binding assay (C1qBA) and polyethylene glycol precipitation (PEG-OD 280).
- Analysis of anti-purified protein derivative antibodies (IgG and IgM) in serum and CICs.
- Western blot analysis of PEG precipitates to characterize complexed antibodies.
Main Results:
- PEG-OD 280 showed significantly elevated CIC levels in tuberculous samples compared to controls.
- Chemotherapy initially increased CIC levels, followed by a decrease with bacterial clearance.
- Antibodies against antigens < 70 kDa within CICs were highly discriminatory for pulmonary tuberculosis (100% in S+C+, 70% in S-C-).
Conclusions:
- PEG-OD 280 is a sensitive method for detecting elevated CICs in tuberculosis.
- Antibody profiling within CICs, particularly for antigens < 70 kDa, offers a promising diagnostic marker for pulmonary tuberculosis.
- CICs and their antibody components may reflect disease activity and response to treatment.
Abstract:
Circulating immune complexes (CICs) in serum samples from patients with pulmonary tuberculosis (bacteriologically positive [S+C+] and bacteriologically negative [S-C-]) and controls (NHC) have been measured by using C1q binding assay (C1qBA) and 3.5% polyethylene glycol precipitation and measurement of absorbance at 280 nm (PEG-OD 280). Although C1qBA did not show any difference between tuberculous and normal serum samples, PEG-OD 280 was significantly elevated in tuberculous samples. The effect of chemotherapy on CIC levels was studied. During the treatment, initially (for up to 2 months) there was a rise in CIC levels and later a fall, coinciding with bacterial clearance. Anti-purified protein derivative antibodies of class immunoglobulin G (IgG) and immunoglobulin M were measured in the serum samples and PEG precipitates. Anti-mycobacterial antibody measurement in CICs was more discriminatory between the groups than serum antibody. For characterization of the complexed antibody, the PEG precipitates were used in the Western blot and the patterns were compared. S+C+ CICs contained antibodies for a wide range of antigens ranging from 100 Kd to 10 Kd. However, none of the NHC-CICs contained antibodies for antigens < 70 Kd. Thus, when using the criterion of positivity for antibodies to antigens < 70 Kd as a marker for pulmonary tuberculosis, 24 of 24 (100%) of the S+C+ CICs were positive. Similarly, 11 of 16 (70%) of the S-C- CICs contained antibodies for antigens < 70 Kd.(ABSTRACT TRUNCATED AT 250 WORDS)
More Related Videos
06:14Separation and Fractionation of Cell Wall and Cell Membrane Proteins from Mycobacterium tuberculosis for Downstream Protein Analysis
Published on: September 26, 2025
07:42Fluorescence Assays for the Study of Mycobacterium tuberculosis Interaction with the Immune Receptor SLAMF1
Published on: February 28, 2025
Related Concept Videos
Defense Against Bacterial Pathogens
Phagocytes
Phagocytes are the frontline soldiers of the immune system. They include neutrophils and macrophages. Neutrophils are the most abundant type of white blood cell and are quickly mobilized to the site of infection. Macrophages are larger cells that patrol...
Pulmonary Tuberculosis I
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
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...
Pulmonary Tuberculosis III
The first classification is based on the development of the disease, and it includes the following categories:
Pulmonary Tuberculosis IV
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...
Tuberculosis