Alterations in neurofilament protein(s) in human leprous nerves: morphology, immunohistochemistry and Western immunoblot correlative study.

Using a specific antibody (SMI 31), the state of phosphorylation of high and medium molecular weight neurofilaments (NF-H and NF-M) was studied in 22 leprous and four nonleprous human peripheral nerves by means of immunohistochemistry, sodium dodecyl sulfate-poly acrylamide gel electrophoresis (SDS-PAGE) and Western immunoblot (WB). The results thus obtained were compared with morphological changes in the respective nerves studied through light and electron microscopy. Many of the leprous nerves showing minimal pathology revealed lack of or weak staining with SMI 31, denoting dephosphorylation. Remyelinated fibres stained intensely with SMI 31 antibody. The WB analysis of Triton X-100 insoluble cytoskeletal preparation showed absence of regular SMI 31 reactive bands corresponding to 200 and 150 kDa molecular weight (NF-H and NF-M, respectively) in 10 nerves. Three of the 10 nerves revealed presence of NF protein bands in SDS-PAGE but not in WB. Presence of additional protein band (following NF-M) was seen in four nerves. Two nerves revealed NF-H band but not NF-M band and one nerve showed trace positivity. In the remaining five nerves presence of all the three NF bands was seen. Thus, 77.3% (17/22) of human leprous nerves studied showed abnormal phosphorylation of NF protein(s). The ultrastructural study showed abnormal compaction and arraying of NF at the periphery of the axons in the fibres with altered axon to myelin thickness ratio (atrophied fibres) as well as at the Schmidt-Lantermann (S-L) cleft region. Such NF changes were more pronounced in the severely atrophied axons suggesting a direct correlation. The observed well-spaced NF in the remyelinated fibres under ultrastructural study was in keeping with both intense SMI 31 staining and presence of NF triplet bands seen in WBs in four of leprous nerves that showed a large number of regenerating fibres suggesting reversal of changes with regeneration. Findings in the present study suggest that atrophy, that is, the reduction in axonal calibre and paranodal demyelination, seen in leprous nerves may result from dephosphorylation of NF-H and NF-M proteins.

The collagen of permanently damaged nerves in human leprosy.

The collagen of nerve biopsies from ten leprosy patients was studied by the Picrosirius-polarization method and electron microscopy. It was observed that leprosy promotes a marked increase in nerve collagen content. Despite the changes induced by this disease, the localization of collagen types I and III in the endo-, peri-, and epineurium remains the same as in normal nerves.

Infection of armadillos with Mycobacterium leprae. Ultrastructural studies of peripheral nerve.

Peripheral nerves of armadillos were studied 16 to 30 months after intradermal or intravenous inoculation with Mycobacterium leprae. Numerous bacilli were found within macrophages, Schwann cells, and perineurial cells; endothelial cells, pericytes and fibroblasts were involved as well. The bacilli were characteristically contained in membrane-limited vacuoles that were interpreted as being phagosomes. Some of the phagosomes contained granular, membranous, and vesicular debris considered to be bacillary degradation products, suggesting that lysosomal activity was present within the phagosomes. Multivesicular bodies, a few of which contained bacilli, were abundant in macrophages and perineurial cells. An unusual proliferation of irregular tubulovesicular profiles was noted, especially in Schwann and perineurial cell cytoplasm, surrounding and within phagosomes containing bacilli. The pattern of cellular involvement of neural structures with M leprae was similar to that observed in lepromatous leprous neuritis in humans.