This survey was to elucidate the bacterial clearance time (BCT) among the patient of L-type and B-group leprosy who had been und.er chernotherapy and foll- ow up with bacterial examination 4 times every year more than 3 years at the Chronic Disease Laboratory of Catholic Medical College. The 99 patients, 53 L-type and 46 B-group, were slected for this study. These patients were classified again into 2 groups: 80 patients with negative bacterial index and 19 patient of posit.ive bacterial index. The patient with negative bacterial index was subdivided into 3 groups by BCT. 1. Rapid decrease group (RA group) BCT<(4years) 2. Standard decrease group (ST group) 4BCT<7 R. Slow decrease group (SL group) 7.
Chronic Disease ; Drug Therapy* ; Humans ; Leprosy* ; Leprosy, Borderline* ; Leprosy, Lepromatous* ; Prognosis*

BACKGROUND: The recent experimental observations suggested that location of M. leprae in the Schwann cells was mediated by epineurial and endoneurial endothelial cells, giving M. leprae access to the inner compartment of nerves and thus to Schwann cells. CD1 is a family of nonpolymorphic beta2-microglobulin-associated transmembrane glycoproteins that is structurally related to classical MHC Ag-presenting molecules, but is encoded by a separate genetic locus. Recent reports have described that human T cells specifically recognize foreign lipid and glycolipid antigens presented by CD1 proteins. Thus, it appeared likely that CD1 represents the key component of a MHC - independent pathway for antigen presentation to T cells. OBJECTIVE: we observed expression of antigen presenting molecules, such as MHC class I, II and CD1, in the vascular endothelial cells and inflammatory cells of leprosy skin lesion. METHODS: MHC class I, II and CD1 expression were studied using immunohistochemical stains. RESULTS: 1. In immunohistochemical stain of HLA-A, B, C, the level of expression in vascular endothelial cells of the borderline tuberculoid leprosy is higher than that of borderline lepromatous leprosy, but lower than that of normal skin tissue. 2. In HLA-A,B,C expression of the inflammatory cells, the level of borderline tuberculoid leprosy is higher than that of borderline lepromatous leprosy and of normal skin tissue(p>0.05). 3. Expression levels of HLA-DP, DQ, DR on endothelial cells decrease significantly in order of normal tissue, borderline tuberculoid leprosy, borderline lepromatous leprosy, lepromatous leprosy(p<0.05). 4. Expression levels of HLA-DP, DQ, DR on inflammatory cells decrease in order of lepromatous leprosy, borderline lepromatous leprosy, borderline tuberculoid leprosy, normal tissue, but statistical significance did not exist. 5. In immunohistochemical stains of CD1b, 3 sections of all 4 normal skin sections and 1 section of 3 borderline tuberculoid leprosy sections showed focal positivity on the dermal inflammtory cells, but borderline lepromatous leprosy sections did not show any positive inflammatory cells. 6. Epidermal Langerhans cells showed positivity on immunohistochemical stains of CD1a and CD1b. CONCLUSION: These results suggest that expression of MHC class I and II on the vascular endothelial cells and expression of CD1b on the inflammatory cells decrease in order of immunity of lepromatous skin lesion and that vascular enothelial cells play an important role in the pathogenesis of leprosy.
Antigen Presentation ; Coloring Agents ; Endothelial Cells* ; Genetic Loci ; Glycoproteins ; HLA-A Antigens ; HLA-DP Antigens ; Humans ; Langerhans Cells ; Leprosy* ; Leprosy, Borderline ; Leprosy, Lepromatous ; Leprosy, Multibacillary ; Leprosy, Paucibacillary ; Leprosy, Tuberculoid ; Schwann Cells ; Skin* ; T-Lymphocytes

BACKGROUND: The causative agents of leprosy are the well-known Mycobacterium leprae and the newly discovered Mycobacterium lepromatosis. This agent was found in 2008, and it was found to be the cause of diffuse lepromatous leprosy in two Mexican patients. OBJECTIVE: The objective of this work was to determine if M. leprae and M. lepromatosis were present in formalin-fixed and paraffin-embedded skin samples from cases from different regions in Mexico. METHODS: A total of 41 skin samples were obtained from 11 states of Mexico. All patients' samples were diagnosed by clinical and histopathological analyses. Total DNA was isolated using a Qiagen-DNeasy blood and tissue kit and molecular identification was achieved by two semi-nested polymerase chain reactions. RESULTS: The 41 patient included 33 samples from men and 8 samples from women; 29 samples were polymerase chain reaction (PCR)-positive to Mycobacterium and 12 samples were PCR-negative. From those 29 samples, 13 were PCR-positive to M. leprae, 8 to M. lepromatosis and 8 were positive to both species. The histopathological diagnosis included; Nodular lepromatous leprosy (NLL); Diffuse lepromatous leprosy (DLL); and Borderline leprosy (BL). The 29 PCR-positive samples were classified as follow: 14 NLL, 4 DLL, and 11 BL. In the 12 samples negative to Mycobacterium, 7 showed the NLL, 2 DLL and 3 BL. CONCLUSION: These findings add evidence to the M. leprae and M. lepromatous distribution, clinical forms and participation of dual infections in Mexico.
Diagnosis ; DNA ; Female ; Hospital Distribution Systems ; Humans ; Leprosy ; Leprosy, Borderline ; Leprosy, Lepromatous ; Male ; Mexico* ; Mycobacterium leprae* ; Mycobacterium* ; Polymerase Chain Reaction ; Skin*

BACKGROUND: PCR from paraffin-embedded tissues has been recently applied on the diagnosis of leprosy, PCR is a highly sensitive technique and the amplified product is detected by gel electrophoresis and Southern blot hybridization. But conventional PCR does not give an information about histopathologic features. On the other hand in situ PCR informs the histopathological location of DNA amplified inside the cells and detected by in sita hybridization or immunohistoche mical method. OBJECTIVE: In situ PCR was applied on the paraffin-embedded tissues of borderline leprosy patients. The optimal condition of in situ PCR in leprosy was searched with the parameters of pretreatment of the tissues, fixation time of paraformaldehyde and total volume of PCR. METHODS: The paraffin-embedded tissues of ten borderline leprosy patients were subjected to in situ PCR. Amplified DNA product within the cells was incorporated with Digoxigenin and was directly detected by immunohistochemical method using anti-Dig-alkaline phosphatase conjugated antibody. The tissues were pretreated with 0.2N HCl or various concentration of proteinase K such as 10, 15 and 100ug/ml and various incubation time of proteinase K from 3.5 minutes to 15 minutes. Fixation was performed before pretreatment. and after PCR for 15 minutes, after pretreatment and after PCR for 15 minutes, only after pretreatment for 15 minutes or after pretretmant for 60 minutes. The total volume of PCR was 40, 50 or 60ul. RESULTS: 1. The amplified DNA was detected using the HCl-pretreated tissues in four of seven BL patients and one of three BT pat ients. 2. The signals were detected in the cytoplasm of most histiocytes and proliferated Schwann cells, some secretory cells of sweat glands and a few endothelial cells in the tissues of the BL patients and the cells composing of the granuloma in those of the BT patients. 3. Proteinase K-treated tissues showed positive reaction in only one tissue used in 10ug/ml of proteinase K for 10 minutes. 4. The proper time for fixation of paraformaldehyde was before treatment of proteinase K or HCl and after PCR reaction. 5. The 40ul of total volume for PCR reaction was enough and minimized the loss of tissues during PCR reaction. CONCLUSION: When in situ PCR was applied on the paraffin-ernbedded tissues of borderline leprosy patients, pretreatment such as concentration and incubation time of proteinase K or HCL was the most critical parameter.
Blotting, Southern ; Cytoplasm ; Diagnosis ; Digoxigenin ; DNA ; Electrophoresis ; Endopeptidase K ; Endothelial Cells ; Granuloma ; Hand ; Histiocytes ; Humans ; Leprosy ; Leprosy, Borderline* ; Polymerase Chain Reaction* ; Schwann Cells ; Sweat Glands

Hansen's disease(HD) is a chronic infectious disorder acquired by inoculation of Mycobacterium leprae. With the establishment of complex multidrug therapy, the incidence rate of leprosy patients has continually shown to decline by 90% compared to the incidence rate in the 1990s. However, the prevalence of the disease still remains high in southeast asian countries. Due to the rarity and diverse nature of cutaneous presentation, HD is often misdiagnosed with other dermatoses or infectious conditions. Especially, when a patient presents with unusual presentation with leprosy reaction with no classical feature such as sensory disorders and skin lesion, the diagnosis is further delayed with misguided treatments. Herein we present a 27-year-old Indonesian immigrant who displayed clinical features mimicking that of orbital cellulitis who was later diagnosed with borderline lepromatous leprosy through histologic and PCR confirmation, in light of alerting the probability of leprosy in immigrants with intractable skin presentations.
Adult ; Asian Continental Ancestry Group ; Diagnosis ; Emigrants and Immigrants ; Humans ; Incidence ; Leprosy ; Leprosy, Borderline ; Leprosy, Multibacillary ; Mycobacterium leprae ; Orbit ; Orbital Cellulitis ; Polymerase Chain Reaction ; Prevalence ; Sensation Disorders ; Skin ; Skin Diseases