Contact-dependent demyelination by Mycobacterium leprae in the absence of immune cells.

Demyelination results in severe disability in many neurodegenerative diseases and nervous system infections, and it is typically mediated by inflammatory responses. Mycobacterium leprae, the causative organism of leprosy, induced rapid demyelination by a contact-dependent mechanism in the absence of immune cells in an in vitro nerve tissue culture model and in Rag1-knockout (Rag1-/-) mice, which lack mature B and T lymphocytes. Myelinated Schwann cells were resistant to M. leprae invasion but undergo demyelination upon bacterial attachment, whereas nonmyelinated Schwann cells harbor intracellular M. leprae in large numbers. During M. leprae-induced demyelination, Schwann cells proliferate significantly both in vitro and in vivo and generate a more nonmyelinated phenotype, thereby securing the intracellular niche for M. leprae.

Neural targeting of Mycobacterium leprae mediated by the G domain of the laminin-alpha2 chain.

We report that the molecular basis of the neural tropism of Mycobacterium leprae is attributable to the specific binding of M. leprae to the laminin-alpha2 (LN-alpha2) chain on Schwann cell-axon units. Using recombinant fragments of LN-alpha2 (rLN-alpha2), the M. leprae-binding site was localized to the G domain. rLN-alpha2G mediated M. leprae binding to cell lines and to sciatic nerves of dystrophic dy/dy mice lacking LN-alpha2, but expressing laminin receptors. Anti-beta4 integrin antibody attenuated rLN-alpha2G-mediated M. leprae adherence, suggesting that M. leprae interacts with cells by binding to beta4 integrin via an LN-alpha2G bridge. Our results indicate a novel role for the G domain of LN-2 in infection and reveal a model in which a host-derived bridging molecule determines nerve tropism of a pathogen.

Organized nerve culture. I. A technique to study the effect of M. leprae infection.

Neonatal dorsal root ganglia were cultivated in vitro by the technique of Murray. Within a week bundles of organized nerve fibers containing proliferating Schwann cells in different phases of axon association and fibroblast cells destined to become peri- or endoneural cells were obtained, peri- or endoneural cells were obtained. Many of these nerve fibers were myelinated within 3-4 weeks. Such 1 or 2 week old cultures were inoculated with M. leprae, and bacilli were found in the cytoplasm of Schwann cells and fibroblasts, demonstrating that these cells are phagocytic in nature and that it is possible to infect them with M. leprae, Schwann cells, mostly in the free or early association phase, engulfed the bacilli, and this affected their further interaction with the axons and subsequent myelin synthesis.

Organized nerve culture. II. DNA synthesis in Schwann cells in the presence of M. leprae.

Schwann cells that contain M. leprae fail to incorporate DNA precursor, indicating blockage of DNA synthesis. Such a block could lead to no proliferation of Schwann cells, an essential requirement for successful association with axons and consequent myelination.