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Separation and Fractionation of Cell Wall and Cell Membrane Proteins from Mycobacterium tuberculosis for Downstream Protein Analysis

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CELLULAR REACTIONS TO DEFATTED TUBERCLE BACILLI AND THEIR PRODUCTS.

F R Sabin¹, A L Joyner

¹Laboratories of The Rockefeller Institute for Medical Research.

The Journal of Experimental Medicine

|October 30, 2009

Summary

Defatted tubercle bacilli trigger complex cellular responses. Their lipid component, containing an acid-fast hydroxy acid and a non-acid-fast polysaccharide, elicits distinct immune reactions including giant cells and eosinophil infiltration.

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Area of Science:

Immunology
Microbiology
Cell Biology

Background:

Cellular responses to tubercle bacilli are intricate.
Defatted tubercle bacilli exhibit complex immunological characteristics.
Heat-killed whole tubercle bacilli induce similar complex reactions.

Purpose of the Study:

To investigate the specific cellular reactions elicited by defatted tubercle bacilli.
To differentiate the immunological roles of lipid and polysaccharide components of tubercle bacilli.
To understand the mechanisms behind foreign body giant cell formation and eosinophil/neutrophil recruitment.

Main Methods:

Isolation and fractionation of lipids from defatted tubercle bacilli.
Analysis of acid-fast and non-acid-fast properties of isolated components.
Histological examination of tissue reactions, including cell infiltration (eosinophils, neutrophiles) and giant cell formation.

Main Results:

The firmly bound lipid from defatted bacilli is non-acid-fast.
An acid-fast hydroxy acid and a non-acid-fast polysaccharide were identified within the lipid.
The hydroxy acid induced foreign body giant cells and eosinophil infiltration.
Polysaccharides stimulated neutrophile recruitment.

Conclusions:

The cellular reactions to defatted tubercle bacilli are mediated by their distinct lipid and polysaccharide constituents.
The acid-fast hydroxy acid is a key component in inducing granulomatous inflammation and eosinophilia.
Polysaccharides play a role in neutrophile chemotaxis in response to tubercle bacilli.