Secretory phosphatases deficient mutant of Mycobacterium tuberculosis imparts protection at the primary site of infection in guinea pigs
Authors: P Vineel Reddy
Anil K Tyagi
Ramandeep Singh
Neetika Jaisinghani
Priyanka Chauhan
Sheetal Gandotra
Journal: PLoS ONE (2013)
Discipline: Multidisciplinary
Keywords: Mycobacterium bovis Bacille Calmette-Guérin; TB vaccines; M.tuberculosis mutants; guinea pigs; virulence genes
Affiliations: Department of Biochemistry, University of Delhi South Campus, New Delhi, India.

Abstract:  Background: The failure of Mycobacterium bovis Bacille Calmette-Guérin to impart satisfactory protection against adult pulmonary tuberculosis has necessitated the development of more effective TB vaccines. The assumption that the vaccine strain should be antigenically as similar as possible to the disease causing pathogen has led to the evaluation of M.tuberculosis mutants as candidate tuberculosis vaccines. Methods/principal findings: In this study, we have generated a mutant of M.tuberculosis (Mtb∆mms) by disrupting 3 virulence genes encoding a mycobacterial secretory acid phosphatase (sapM) and two phosphotyrosine protein phosphatases (mptpA and mptpB) and have evaluated its protective efficacy in guinea pigs. We observed that Mtb∆mms was highly attenuated in THP-1 macrophages. Moreover, no bacilli were recovered from the lungs and spleens of guinea pigs after 10 weeks of Mtb∆mms inoculation, although, initially, the mutant exhibited some growth in the spleens. Subsequently, when Mtb∆mms was evaluated for its protective efficacy, we observed that similar to BCG vaccination, Mtb∆mms exhibited a significantly reduced CFU in the lungs of guinea pigs when compared with the unvaccinated animals at 4 weeks after challenge. In addition, our observations at 12 weeks post challenge demonstrated that Mtb∆mms exhibited a more sustainable and superior protection in lungs as compared to BCG. However, the mutant failed to control the hematogenous spread as the splenic bacillary load between Mtb∆mms vaccinated and sham immunized animals was not significantly different. The gross pathological observations and histopathological observations corroborated the bacterial findings. Inspite of disruption of phosphatase genes in MtbΔmms, the lipid profiles of M.tuberculosis and MtbΔmms were identical indicating thereby that the phenotype of the mutant was ascribed to the loss of phosphatase genes and the influence was not related to any alteration in the lipid composition. Conclusions/significance: This study highlights the importance of M.tuberculosis mutants in imparting protection against pulmonary TB. 

Retracted: No

PubPeer comments:
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 Fig 4(c).  TEM images of ratsu2019 liver tissues in each group (N, nucleus; M, mitochondrion; L, lipid droplet) . ![file] I see no N, M, L. What do the white arrows represent in some of the panels? They are not mentioned in the text. , 
 Fig 6a.  HE staining images of large aortic tissues of rats in each group . Other panels are left as an exercise for the reader. ![file] ,