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1.
Blood ; 106(5): 1843-50, 2005 Sep 01.
Article in English | MEDLINE | ID: mdl-15886329

ABSTRACT

The early innate response after Mycobacterium bovis bacille Calmette-Guérin (BCG) vaccination is poorly characterized but probably decisive for subsequent protective immunity against tuberculosis. Therefore, we vaccinated mice with fluorescent BCG strains in the ear dorsum, as a surrogate of intradermal vaccination in humans. During the first 3 days, we tracked BCG host cells migrating out of the dermis to the auricular draining lymph nodes (ADLNs). Resident skin dendritic cells (DCs) or macrophages did not play a predominant role in early BCG capture and transport to ADLNs. The main BCG host cells rapidly recruited both in the dermis and ADLNs were neutrophils. Fluorescent green or red BCG strains injected into nonoverlapping sites were essentially sheltered by distinct neutrophils in the ADLN capsule, indicating that neutrophils had captured bacilli in peripheral tissue and transported them to the lymphoid organ. Strikingly, we observed BCG-infected neutrophils in the lumen of lymphatic vessels by confocal microscopy on ear dermis. Fluorescence-labeled neutrophils injected into the ears accumulated exclusively into the ipsilateral ADLN capsule after BCG vaccination. Thus, we provide in vivo evidence that neutrophils, like DCs or inflammatory monocytes, migrate via afferent lymphatics to lymphoid tissue and can shuttle live microorganisms.


Subject(s)
BCG Vaccine/administration & dosage , Cell Movement/immunology , Lymph Nodes/immunology , Lymphatic System/immunology , Mycobacterium bovis/immunology , Neutrophils/immunology , Neutrophils/microbiology , Animals , BCG Vaccine/immunology , Dendritic Cells/immunology , Ear , Female , Fluorescent Dyes/chemistry , Injections, Intradermal , Mice , Mice, Inbred C57BL , Neutrophil Activation , Staining and Labeling , Tissue Distribution
2.
Infect Immun ; 70(5): 2256-63, 2002 May.
Article in English | MEDLINE | ID: mdl-11953357

ABSTRACT

Adaptive regulation of gene expression in response to environmental changes is a general property of bacterial pathogens. By screening an ordered transposon mutagenesis library of Mycobacterium tuberculosis, we have identified three mutants containing a transposon in the coding sequence or in the 5' regions of genes coding for two-component signal transduction systems (trcS, regX3, prrA). The intracellular multiplication capacity of the three mutants was investigated in mouse bone marrow-derived macrophages. Only the prrA mutant showed a defect in intracellular growth during the early phase of infection, and this defect was fully reverted when the mutant was complemented with prrA-prrB wild-type copies. The mutant phenotype was transient, as after 1 week this strain recovered full growth capacity to reach levels similar to that of the wild type at day 9. Moreover, a transient induction of prrA promoter activity was observed during the initial phase of macrophage infection, as shown by a prrA promoter-gfp fusion in M. bovis BCG infecting the mouse macrophages. The concordant transience of the prrA mutant phenotype and prrA promoter activity indicates that the PrrA-PrrB two-component system is involved in the environmental adaptation of M. tuberculosis, specifically in an early phase of the intracellular growth, and that, similar to other facultative intracellular parasites, M. tuberculosis can use genes temporarily required at different stages in the course of macrophage infection.


Subject(s)
Bacterial Proteins/physiology , Mycobacterium tuberculosis/growth & development , Protein Kinases/physiology , Trans-Activators/physiology , Animals , Bacterial Proteins/genetics , DNA Transposable Elements , Female , Histidine Kinase , Macrophages/microbiology , Mice , Mice, Inbred BALB C , Mutation , Mycobacterium tuberculosis/genetics , Protein Kinases/genetics , Trans-Activators/genetics
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