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PubMed1993

Detection of Mycobacterium leprae DNA in formalin-fixed, paraffin-embedded samples from multibacillary and paucibacillary leprosy patients by polymerase chain reaction.

BACKGROUND: Diagnosis of paucibacillary leprosy is often difficult. A method that could confirm the diagnosis is the polymerase chain reaction (PCR) of M. leprae DNA. This reaction was applied to biopsied tissues of leprotic patients to determine the suitability and sensitivity of the reaction. METHODS: Biopsy samples were taken from previously untreated patients with multibacillary (5 patients) and paucibacillary (3 patients) leprosy, fixed in formalin, and embedded in paraffin. DNA was extracted from paraffin blocks and PCR applied. The sensitivity of the PCR method was tested by using the serially diluted DNA sample as the template. RESULTS: All eight patients showed a positive PCR for M. leprae DNA. The sensitivity was such that a single organism of M. leprae, as counted by staining of the acid-fast bacilli was identified by the PCR. CONCLUSIONS: The PCR method is simple, sensitive, specific, and does not require the use of radioisotopes. It can be applied to the unequivocal diagnosis of paucibacillary leprosy which is difficult by other means. The diagnosis can be obtained within 10 hours.

Blotting, Southern↗
PubMed1993

Gallium-67 scintigraphy in borderline lepromatous leprosy.

A middle aged woman with a pyrexia of unknown origin was shown to have borderline lepromatous leprosy. Early gallium-67 scintigraphy demonstrated increased uptake in the subcutaneous tissues of the face and thighs. As a result of these findings skin biopsy was obtained from the right thigh which gave a diagnosis of borderline lepromatous leprosy. The authors have been unable to find other reports of gallium-67 scintigraphy in leprosy but the pattern of gallium-67 distribution should suggest the diagnosis.

Adult↗
PubMed1991

Identification of Mycobacterium leprae antigens from a cosmid library: characterization of a 15-kilodalton antigen that is recognized by both the humoral and cellular immune systems in leprosy patients.

Screening of the Mycobacterium leprae cosmid library with pooled sera from lepromatous leprosy (LL) patients by a colony immunoblot technique resulted in the identification of about 100 colonies that produced immunologically reactive proteins. Twenty-four of these clones were purified, analyzed, and found to comprise two groups according to the reactivity of the recombinant proteins with LL sera and to the DNA restriction patterns of the recombinant plasmids and cosmids. Proteins specified by clones from group I reacted strongly with LL patients' sera on a Western blot (immunoblot), demonstrating a 15-kDa protein band designated A15. The A15 antigen also reacted with pooled sera from patients with tuberculoid leprosy from the United States and Brazil. Clones from group II did not show any reactive protein band on a Western blot, when reacted with patients' sera. DNAs from cosmids of group II all contain a 10-kb PstI fragment that hybridized to the unique repetitive M. leprae DNA. Sequence analysis of a 1.2-kb fragment containing the entire coding sequence of A15 revealed three open reading frames (ORFs), only one of which (ORF II) contains sufficient genetic information to encode for A15. Part of the A15 gene was found to exist also in a group of lambda gt11:M. leprae clones previously isolated in our laboratory by immunological screening with LL patients' sera. One of the lambda gt11 clones (L8) expresses a beta-galactosidase fusion protein with 89 amino acids from the C terminus of A15. An important result was that the fusion protein was clearly recognized by T cells from leprosy patients. Interestingly, Mycobacterium tuberculosis-stimulated T cells from M. leprae nonresponder (LL as well as borderline tuberculoid) patients were able to respond to the isolated recombinant M. leprae antigen, indicating that nonresponsiveness to M. leprae antigens can be reversible. The sequence of the M. leprae DNA fused to the beta-galactosidase gene of lambda gt11 clone L8 was identical to that of a lambda gt11:M. leprae clone isolated recently that expresses an immunologically reactive fusion protein (S. Laal, Y. D. Sharma, H. K. Prasad, A. Murtaza, S. Singh, S. Tangri, R. Misra, and I. Nath, Proc. Natl. Acad. Sci. USA 88:1054-1058, 1991). Besides the complete sequence of the A15 gene, sequencing data of two flanking ORFs are presented. Downstream from ORF II (A15), ORF III has a high degree of similarity to the genes for tomato ATP-dependent proteases that are members of a larger class of highly conserved proteases ubiquitous among prokaryotes and eukaryotes.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗
PubMed1991

Influence of untreated chronic plastic iridocyclitis on intraocular pressure in leprosy patients.

The intraocular pressures of a total of 286 eyes of patients with lepromatous and borderline lepromatous leprosy who never had regular ophthalmological care or local eye treatment were measured. The patients were categorised according to the type of leprosy they had, and the eyes were categorised as without or with chronic plastic iridocyclitis. In patients with lepromatous and borderline lepromatous types of leprosy the intraocular pressure was significantly lower in eyes with chronic plastic iridocylitis 10.1 (3.6) mmHg than in both unaffected eyes 11.0 (3.2) mmHg and control eyes 13.5 (2.5) mmHg. It has been shown that chronic plastic iridocyclitis which remains untreated for years results in a lower intraocular pressure than normal.

Adult↗
PubMedDate not supplied

[Edema in leprosy: the clinical and therapeutic aspects].

Edema, which is commonly described as a symptom of reactional states, may occur during the course of leprosy. Both diagnosis and adequate treatment measures are often difficult to achieve and failure to do so may result in permanent damage to the lower limbs. In a one-year follow-up study of leprosy patients--ten multibacillary and one paucibacillary--who had been submitted to a clinical protocol for diagnosis and pathological classification, a clinical pattern of localized and/or systemic edema was observed. Among these patients, five simultaneously presented other symptoms related to reactional states, 4 were diagnosed as Type I, and one as Type II. On the other hand, while three of the patients did not present reaction at the time when edema was diagnosed, they did develop some aspects of reactional disease later on (two had neuritis e one had Type I reaction). The edemas that preceded or were associated with reactional episodes showed clinical regression as a result of specific treatment against reactions (corticosteroids and/or pentoxifylline and/or thalidomide) in the absence of another treatment normally used for edemas. Although these data need to be confirmed by controlled studies, they strongly suggest that immunological mechanisms are involved in the physiopathology of edema in leprosy.

Adolescent↗
PubMed2000

A role for CD40-CD40 ligand interactions in the generation of type 1 cytokine responses in human leprosy.

The interaction of CD40 ligand (CD40L) expressed by activated T cells with CD40 on macrophages has been shown to be a potent stimulus for the production of IL-12, an obligate signal for generation of Th1 cytokine responses. The expression and interaction of CD40 and CD40L were investigated in human infectious disease using leprosy as a model. CD40 and CD40L mRNA and surface protein expression were predominant in skin lesions of resistant tuberculoid patients compared with the highly susceptible lepromatous group. IL-12 release from PBMC of tuberculoid patients stimulated with Mycobacterium leprae was partially inhibited by mAbs to CD40 or CD40L, correlating with Ag-induced up-regulation of CD40L on T cells. Cognate recognition of M. leprae Ag by a T cell clone derived from a tuberculoid lesion in the context of monocyte APC resulted in CD40L-CD40-dependent production of IL-12. In contrast, M. leprae-induced IL-12 production by PBMC from lepromatous patients was not dependent on CD40L-CD40 ligation, nor was CD40L up-regulated by M. leprae. Furthermore, IL-10, a cytokine predominant in lepromatous lesions, blocked the IFN-gamma up-regulation of CD40 on monocytes. These data suggest that T cell activation in situ by M. leprae in tuberculoid leprosy leads to local up-regulation of CD40L, which stimulates CD40-dependent induction of IL-12 in monocytes. The CD40-CD40L interaction, which is not evident in lepromatous leprosy, probably participates in the cell-mediated immune response to microbial pathogens.

CD40 Antigens↗
PubMed2002

[Leprosy reactions].

In leprosy, the causative bacteria, Mycobacterium leprae, will not threaten the lives of the hosts directly because they proliferate only slowly in the Schwann cells of the peripheral nerves. It is the "reactions" which give the patients irreversible morbidity through the inflammatory damages to the peripheral nerves. Physicians should be aware of the possibility of the state of the "reaction" when they examine leprosy patients. They also should be aware of the possibility of leprosy and the state of the "reaction" when they examine patients with cutaneous lesions and/or peripheral nerve disturbances, because it may be the first presenting symptom of the disease. In this review, recent advances on the issue about the reactions are discussed including pathogenesis, immunology, clinical features, pathology, treatment and prevention.

Female↗
PubMedDate not supplied

A florid case of borderline-lepromatous leprosy.

A case of borderline-lepromatous leprosy is presented to highlight the detection and prevalence of florid, new cases of leprosy towards the lepromatous end of the spectrum in the northern and western regions of Australia; the recognition and early treatment of lepra reactions; and the potential use of an enzyme-linked immunosorbent assay to detect and to monitor immunoglobulin M levels in leprosy.

Adult↗
PubMed1989

Ocular changes in reactions in leprosy.

A study of ocular changes in reactions in leprosy was undertaken to assign these changes, their proper place in the wide spectrum of ocular morbidity in leprosy. 76.1% of eyes of Type I reaction and 89.7% of eyes with Type II reaction showed some ocular involvement. Corneal hypoaesthesia, superficial punctate keratitis, a decrease of corneal film break up time (BUT), prominent corneal nerves, pigment on the endothelium of the cornea and a pigmented trabecular meshwork were the common ocular findings. The incidence of iridocyclitis in Type II reactions was low (8.1%). The significance of the ocular involvement in reactions in leprosy and the pathogenesis of iridocyclitis in Type II reactions is discussed.

Adolescent↗
PubMed1990

A study of relapse in paucibacillary leprosy in a multidrug therapy project, Baroda District, India.

In order to judge the value of therapeutic regimens in paucibacillary leprosy, knowledge of incubation time of relapses is essential, as this will define the length of time patients have to be followed up after treatment has been stopped. The prospective study of relapse includes paucibacillary cases of leprosy belonging to a non-lepromatous group consisting of tuberculoid, neuritic and indeterminate. Data are presented on the incubation time of 21 relapses after multidrug therapy in Baroda district; 76.19% of relapses occur during the first 2 years. This figure is most important in the analysis of results of drug trials in paucibacillary leprosy. This figure should also be relevant to regimens including drugs that are more bacteriocidal than dapsone, since the bacteriocidal activity has a bearing on the minimal necessary duration of treatment, but not on the incubation time of relapses. With the introduction of bactericidal drugs e.g. rifampicin in multidrug therapy, the incidence of relapse are very low, hence relapse rates fall down to a very low level after multidrug therapy. Our study shows a mean relapse rate of 0.19% after multidrug therapy. Factors associated with the occurrence of relapse are discussed.

Adolescent↗
PubMed2000

Dysregulation of IL-4 expression in lepromatous leprosy patients with and without erythema nodosum leprosum.

In order to increase our understanding of the immunological basis of erythema nodosum leprosum (ENL), we studied Th-like cytokine profiles in 130 leprosy patients, employing both the conventional and a novel, real-time, fluorogenic reverse transcriptase-based PCR (RT-PCR). The concomitant expression of both Th-like cytokines, interferon-gamma and IL-4, and the regulatory cytokines, IL-10 and IL-12, was studied in the peripheral blood cells of leprosy patients with and without ENL. In the conventional RT-PCR, varied cytokine profiles were observed in individual patients of all clinical types. Fifty-three percent of lepromatous patients without ENL and 59% of tuberculoid leprosy patients showed co-expression of IFN gamma and IL-4, indicating a non-polarized Th 0 pattern. Of the 36 patients with ENL, 58% demonstrated a polarized Th 1 pattern, with only 30% expressing both cytokines. Semiquantitative RT-PCR indicated a lower expression of IL-4 compared to that of IFN gamma in the lepromatous patients without ENL; the difference was even greater among those with ENL. The sensitive, real-time PCR confirmed the down-regulation of IL-4 and IL-10, with absence of IL-4 in half of the patients, resulting in skewing of the cytokine response toward a Th 1-like profile.

Cytokines↗
PubMed1999

Serum IL-1ra is elevated in lepromatous leprosy patients.

Patterns of production of specific cytokines are accepted as standards for T-lymphocyte subsets in diseases caused by intracellular parasites. These lymphocyte subsets (Th1 and Th2) have been associated with the different poles of the leprosy spectrum. Lepromatous leprosy (LL) onset correlates with cytokines produced by Th2 cells on the grounds of the patient's poor cellular immune response, i.e., interleukin 2 (IL-2) and gamma interferon (IFN-gamma) deficiency. On the other hand, tuberculoid leprosy (TL) has been associated with a Th1 response. Moreover, pro-inflammatory cytokines like IL-1 beta and tumor necrosis factor-alpha (TNF-alpha) play a major role in chronic inflammatory pathologies being IL-1ra and TNF-alpha soluble receptors, natural counterbalancing inhibitors. In light of this background, we decided to measure serum levels of IL-1 beta, IL-1ra, TNF-alpha and IL-6 in LL and TL patients, and we also studied the production in vitro of Th1 (IFN-gamma, IL-2), Th2 (IL-4, IL-10) and TNF-alpha cytokines. Our data showed that IL-1ra is highly elevated in sera from LL patients; there were no differences in Th2 cytokine levels and there were diminished levels in Th1 cytokines.

Adult↗
PubMed2000

[A rare cause of carpal tunnel syndrome: leprosy. Latency period of decades is possible].

In a 63-year-old German woman with no apparent exposure to leprosy, clinical and histological evidence of tuberculoid leprosy was found. A noteworthy feature of this case is either the very long incubation time of 45 years, or the mode of infection involving an only short encounter in an endemic region. In the event of unclear lesions--on white skin not, typically, hypopigmented, but reddish--accompanied by disordered sensation, we in Europe must also give consideration to the possibility of a sporadic case of leprosy. A particular characteristic of this case was leprous neuritis involving the median nerve which was diagnosed clinically as carpal tunnel syndrome, and scheduled for surgical treatment. Under treatment with dapsone and rifampicin, however, the condition cleared up completely.

Carpal Tunnel Syndrome↗
PubMedDate not supplied

Thyroid functional status in leprosy patients in Sudan.

We studied 45 adult patients with untreated lepromatous leprosy and borderline leprosy, presenting at clinics in Khartoum and Omdurman, to assess clinical and biochemical effects of the disease on thyroid function. A matching control group of 30 subjects, without symptoms or signs of thyroid disease, were included for comparison. Thyroxine, triiodothyronine and thyrotrophin levels were within normal range. Mean serum thyroxine was low in both groups (significant in lepromatous leprosy patients only). Mean serum triiodothyronine was high in both groups (significant in neither group). Mean thyrotrophin was significantly higher in both groups compared with controls.

Adolescent↗
PubMedDate not supplied

Incidence of leprosy in school-children and their family members in Berhampur.

A school survey, followed by a contact survey, was carried out in Berhampur, a city in southern Orissa. In a study of 8,870 school-children, leprosy was detected in 15, giving a prevalence rate of 16.91 per 10,000 with a male:female ratio of 8:7. Of these, 14 (93.99%) had paucibacillary leprosy. More cases [11 (73.33%)] were seen in the age-group of 10-15 years. Exposed parts, such as lower limbs, upper limbs and head and neck in that order, were the sites of predilection, accounting for 85.71% of total lesions. Nerve involvement was found in 2 (13.33%) girls with deformity (ulnar claw) in one of them (6.66%). BCG scar was present in 11 (73.33%) cases. Among the vaccinated cases, tuberculoid type was the most common, followed by indeterminate, pure neuritic and borderline, in that order. A contact survey detected 2 multibacillary cases in two families (13.33%). In each case, the father was the index source. The study revealed that a maximum number of students, 8 (53.3%), belonged to the middle socioeconomic class. Of the 15 affected, 60% were undernourished and the rest well nourished. No other systemic disease was found clinically associated with leprosy.

Adolescent↗
PubMedDate not supplied

An ultrastructural study of Schwann cells in peripheral nerves of leprosy patients.

An ultrastructural study of peripheral nerves in leprosy patients was carried out of ascertain the changes in Schwann cells containing myelinated and nonmyelinated axons. Axonal multiplication was noticed in nonmyelinated axons in specimens from both tuberculoid and lepromatous leprosy. The Schwann cells in tuberculoid nerves were devoid of M. leprae in contrast to those in lepromatous nerves in which large number of bacilli were seen. These observations suggest that the Schwann cells containing nonmyelinated axons may be affected more frequently in either type of leprosy.

Axons↗
PubMedDate not supplied

Use of reverse transcription polymerase chain reaction for the detection of Mycobacterium leprae in the slit-skin smears of leprosy patients.

The relevance of bacterial index (BI) for understanding the prognosis of leprosy patients on treatment has been extensively debated, as it does not give a very clear idea of the viability of the bacteria in patients under treatment. Here we used slit-skin smear samples of leprosy patients to test the suitability for studying viability of Mycobacterium leprae using reverse transcription polymerase chain reaction (RT-PCR). For this purpose, we recruited 13 multibacillary (MB) leprosy patients (8 lepromatous and 5 borderline lepromatous). Of these, 7 were relapse cases, 3 were under treatment (MB-MDT), 2 were new cases and 1 had completed treatment. We carried out extraction of RNA using Trizol reagent (Life Technologies, UK) from the slit-skin smear samples from these patients. The RNA preparation was then used for the RT-PCR using Mycobacterium leprae-specific primers for the fragment of 16s ribosomal RNA gene. Samples from both the new cases, 4 suspected relapse cases and 1 patient under treatment showed positive RT-PCR results. Other 6 patients whose smear samples did not show any amplification by RT-PCR were on MB-MDT from 8 to 30 months. The usefulness of the technique needs to be validated using mouse footpad technique and also should be more extensively explored for studying the viability of M. leprae, the efficacy of treatment and the presence of other mycobacteria in the slit-skin smear samples.

Adult↗
PubMed1991

Quantitation of the soluble E-receptor of human T lymphocytes by rocket electrophoresis in the serum of patients with lepromatous and tuberculoid leprosy.

Human T lymphocytes carry a membrane receptor for sheep erythrocytes (E) which is responsible for the well-known phenomenon of E-rosette formation. This receptor has been related to CD2 molecules; it is present in a soluble form (Rs) in normal serum and may play an immunoregulatory role. In this study we quantitated soluble E-receptor in serum samples of 43 normal controls, 32 patients with tuberculoid leprosy and 53 with lepromatous leprosy, using rocket electrophoresis and an anti E receptor serum (anti-Rs) obtained from an adult sheep immunized with autologous E treated with Rs. In the 3 groups studied, the rocket means were respectively 5.0, 7.5 and 10.9 mm (p less than 0.001). We found abnormally high levels of Rs in the serum of various diseases associated with a depression of cell-mediated immunity. The increase of Rs levels in the serum may be one of the mechanisms responsible for the depression of cellular immunity in leprosy.

Adult↗