ABSTRACT
The nerve biopsies of 11 patients with pure neuritic leprosy were submitted to routine diagnostic procedures and immunoperoxidase staining with antibodies against axonal (neurofilament, nerve growth factor receptor (NGFr), and protein gene product (PGP) 9.5) and Schwann cell (myelin basic protein, S-100 protein, and NGFr) markers. Two pairs of non-adjacent histological cross-sections of the peripheral nerve were removed for quantification. All the fascicles of the nerve were examined with a 10X-ocular and 40X-objective lens. The immunohistochemistry results were compared to the results of semithin section analysis and clinical and electroneuromyographic data. Neurofilament staining was reduced in 100 percent of the neuritic biopsies. NGFr positivity was also reduced in 81.8 percent, PGP staining in 100 percent of the affected nerves, S100 positivity in 90.9 percent, and myelin basic protein immunoreactivity in 90.9 percent. Hypoesthesia was associated with decreased NGFr (81.8 percent) and PGP staining (90.9 percent). Reduced potential amplitudes (electroneuromyographic data) were found to be associated with reduced PGP 9.5 (63.6 percent) and nerve fiber neurofilament staining (45.4 percent) by immunohistochemistry and with loss of myelinated fibers (100 percent) by semithin section analysis. On the other hand, the small fibers (immunoreactive dots) seen amid inflammatory cells continued to be present even after 40 percent of the larger myelinated fibers had disappeared. The present study shows an in-depth view of the destructive effects of leprosy upon the expression of neural markers and the integrity of nerve fiber. The association of these structural changes with the clinical and electroneuromyographic manifestations of leprosy peripheral neuropathy was also discussed.
Subject(s)
Adult , Female , Humans , Male , Antigens, Bacterial/analysis , Glycolipids/analysis , Leprosy/diagnosis , Mycobacterium leprae/immunology , Nerve Fibers, Myelinated/pathology , Nerve Tissue Proteins/analysis , Neuritis/diagnosis , Antigens, Bacterial/immunology , Biopsy , Biomarkers/analysis , DNA, Bacterial/analysis , Electromyography , Glycolipids/immunology , Immunoenzyme Techniques , Immunohistochemistry , Leprosy/pathology , Myelin Basic Protein , Mycobacterium leprae/genetics , Neuritis/pathology , Neurofilament Proteins/analysis , Polymerase Chain Reaction , Receptors, Nerve Growth Factor/analysis , /analysisABSTRACT
Celulas peritoneais de camundongos cultivadas in vitro foram infectadas com inoculos identicos de amastigotas (AM) e de promastigotas, respectivamente, virulentas (VP), avirulentas (AVP) e fixadas (FP) de uma mesma cepa de Leishmania m. mexicana.As monocamadas coradas com May-Grunwald-Giemsa foram examinadas em diferentes intervalos de tempo. Ao nivel de 3a. hora pos-infeccao o numero de macrofagos contendo parasitas variou entre 60,5% (VP) e 84% (AM) nas monocamadas expostas aos parasitas viaveis, comparados com 6,5% para aquelas expostas aos parasitas fixados.Entretanto, nesse tempo de interacao, houve alteracoes degenerativas parasitarias e o numero de macrofagos contendo parasitas intatos variou significantemente entre as celulas infectadas com AM (84%) e aquelas infectadas com VP (42%) ou AVP (40%). O numero medio de parasitas intatos/macrofago tambem diferiu significantemente entre as celulas infectadas com AM e aquelas infectadas com promastigotas viaveis ou fixadas. A analise quantitativa de parasitas intatos e degenerados indicou uma multiplicacao e destruicao parasitaria nas monocamadas infectadas com VP e sobrevivencia e multiplicacao parasitarias naquelas infectadas com AM. Em contraste, nao houve evidencia de multiplicacao de parasitas nas celulas infectadas com AVP. Exses resultados sugerem que os eventos cruciais que determinam a evolucao da infeccao ocorrem durante as primeiras 24 horas da interacao parasito-hospedeiro. Esses eventos sao aparentemente influenciados nao somente pela cepa do parasita ou do hospedeiro, mas tambem por variacoes ambientais induzidas em uma dada cepa parasitaria