Robert Lee Hunter1, Nandagopal Venkataprasad, Margaret R Olsen. 1. Department of Pathology and Laboratory Medicine, University of Texas-Houston Medical School, MSB 2.136, 6431 Fannin, Houston, TX 77030, USA. Robert.L.Hunter@UTH.TMC.EDU
Abstract
SETTING: M. tuberculosis (MTB) lose virulence during prolonged culture on artificial media. This loss of virulence is associated with a change in colony morphology. Several studies suggested that trehalose 6,6' dimycolate (TDM or cord factor), contributes to colony morphology. OBJECTIVE: To investigate the role of TDM in colony morphology of MTB using clinical isolates selected to have colony morphology typical of virulent or attenuated organisms. DESIGN: Use immunohistochemical and physical chemical methods to assess the presence and distribution of TDM in rapidly growing pellicles of MTB. RESULTS: TDM forms an insoluble crystalline monolayer at the air-water interfaces that is more rigid than that formed by any other biologic amphiphile and is strong enough to support a spreading pellicle of MTB. The surface of young pellicles of the isolate with virulent morphology displayed the regular linear pattern characteristic of monolayers of TDM. TDM was also identified in the open spaces of pellicles of MTB by immunohistochemistry. MTB with morphology of attenuated organisms had neither of these properties. CONCLUSION: These data suggest that the characteristic morphology of colonies of virulent MTB is due to TDM released from the surface of the organisms.
SETTING:M. tuberculosis (MTB) lose virulence during prolonged culture on artificial media. This loss of virulence is associated with a change in colony morphology. Several studies suggested that trehalose 6,6' dimycolate (TDM or cord factor), contributes to colony morphology. OBJECTIVE: To investigate the role of TDM in colony morphology of MTB using clinical isolates selected to have colony morphology typical of virulent or attenuated organisms. DESIGN: Use immunohistochemical and physical chemical methods to assess the presence and distribution of TDM in rapidly growing pellicles of MTB. RESULTS: TDM forms an insoluble crystalline monolayer at the air-water interfaces that is more rigid than that formed by any other biologic amphiphile and is strong enough to support a spreading pellicle of MTB. The surface of young pellicles of the isolate with virulent morphology displayed the regular linear pattern characteristic of monolayers of TDM. TDM was also identified in the open spaces of pellicles of MTB by immunohistochemistry. MTB with morphology of attenuated organisms had neither of these properties. CONCLUSION: These data suggest that the characteristic morphology of colonies of virulent MTB is due to TDM released from the surface of the organisms.
Authors: Thomas R Lerner; Christophe J Queval; Rachel P Lai; Matthew Rg Russell; Antony Fearns; Daniel J Greenwood; Lucy Collinson; Robert J Wilkinson; Maximiliano G Gutierrez Journal: JCI Insight Date: 2020-05-21