Mycobacterium Leprae Research Articles

A case report and literature review: Mycobacterium leprae infection diagnosed by metagenomic next-generation sequencing of cerebrospinal fluid

BackgroundLeprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae) that is responsible for deformities and irreversible peripheral nerve damage and has a broad spectrum of clinical and serological manifestations. Leprosy primarily affects the peripheral nerves and rarely presents with central nervous system involvement. Diagnosing leprosy can still be difficult in some cases, especially when the infection involves uncommon clinical manifestations and extracutaneous sites. Delayed diagnosis and treatment of leprosy may lead to irreversible damage and death.Case PresentationWe report a case of a 30-year-old female presenting with “repeated high fever with symptoms of headache for 14 days”. On the day of admission, physical signs of lost eyebrows and scattered red induration patches all over her body were observed. The patient’s diagnosis was based on the clinical characteristics using a combination of metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF) and slit-skin smear. After confirming Listeria meningitis and multibacillary leprosy with erythema nodosum leprosum (ENL), a type 2 reaction, she was treated with ampicillin sodium, dapsone, rifampicin, clofazimine, methylprednisolone, and thalidomide. At the 1-year follow-up, the frequency and severity of headaches have significantly decreased and a good clinical response with improved skin lesions was found.ConclusionThis case highlights the importance of considering leprosy, which is a rare and underrecognized disease, in the differential diagnosis of skin rashes with rheumatic manifestations, even in areas where the disease is not endemic, and physicians should be alerted about the possibility of central nervous system infections. In addition, mNGS can be used as a complementary diagnostic tool to traditional diagnostic methods to enhance the diagnostic accuracy of leprosy.

Molecular Identification of Mycobacterium leprae in the Leprosy Patients.

Several discoveries about leprosy indicate that Mycobacterium leprae transmission mainly occurs by inhalation, and the nose is a major port of entry and exit. Molecular probes have shown certain potential for the detection and identification of M. leprae in patients. The aim of this study was to identify M. leprae in nasal swab specimens using polymerase chain reaction (PCR)-based assays followed by gene sequencing methods. This observational study examines 64 anterior nasal swab samples taken from pretreatment leprosy patients, on-treatment and completed leprosy treatment in Bulukumba, South Sulawesi, Indonesia. samples were analyzed by molecular detection methods according to the standard methods at the Clinical Microbiology Laboratory of Hasanuddin University. Descriptive statistics were utilized to summarize patient demographics and outcomes. This study uses PCR to detect the M. leprae deoxyribonucleic acid (DNA) from nasal swab specimens. Data were collected from 64 patients with a percentage of male patients 51.54%. Based on the age category, the group 45-46 years was the most frequent (39.05%). PCR detection proline-rich antigen gene of a 531 bp DNA fragment from M. leprae, was positive in eight patients, and they were multibacillary. Furthermore, PCR was positive in 5 (31.25%) of 16 new leprosy patients, 2 (8.69%) of 23 on-treatment patients, and 1 (4%) of 25 treatment completed patients. Based on the results of the phylogenetic tree and analysis of 8 positive results detected by M. leprae from leprosy patients, almost all samples have a level of similarity, except for sample Ua7. M. leprae cannot grow in vitro, so molecular diagnostic tools were used to confirm the disease. This study predominantly of males with the age above 45 years of age being the most common. Eight M. leprae were positive from nasal swab leprosy patients. The sequencing findings provide insight into the genetic diversity of the genus M. leprae, so it is necessary to consider the detection of whole-genome sequence.

Possible Mechanism of Lipid Uptake by Mycobacterium leprae in Host Cells

Possible Mechanism of Lipid Uptake by Mycobacterium leprae in Host Cells

Prevalence of Mycobacterium leprae and Mycobacterium lepromatosis in roadkill armadillos in Brazil

To evaluate the prevalence of Mycobacterium leprae and Mycobacterium lepromatosis in road killed armadillos identified along Brazilian regions, samples of liver, spleen, muscle, ear, nose and tail were collected on highways from 78 animals. The armadillos were of four different species, Cabassous tatouay, Dasypus novemcinctus, Dasypus septemcinctus and Euphractus sexcinctus. After DNA extraction from two tissues, specific primers were used for the detection of each pathogen using SYBR green qualitative Real-Time PCR, and amplicons were sequenced. The species with the highest prevalence was D. novemcinctus, mainly in the Central-West, South, and Southeast regions of Brazil. We detected M. leprae DNA in 32 (41 %) of the 78 individuals and M. lepromatosis DNA was not identified in any of the examined samples. The zoonotic component of leprosy may play a role in the transmission of the disease in endemic areas in which environmental conditions and contact with reservoirs must be investigated.

Clinical and epidemiological panorama of leprosy at a tertiary center across the Nepal–India border in the post-elimination era: A cross-sectional study

ackground: Leprosy, an infectious granulomatous disease caused by Mycobacterium leprae continues to pose a significant public health challenge globally. Despite efforts towards its elimination, leprosy remains a re-emerging threat, with increasing cases in Nepal and India. Understanding the clinical, epidemiological, socioeconomic, and histological profiles of leprosy across the Nepal–India border is essential for effective disease management and control. Methods: A descriptive, prospective study was conducted over three years at a tertiary care center in Nepal. The patients were selected based on predefined inclusion and exclusion criteria. Clinical, epidemiological, and socioeconomic data were collected and analyzed to evaluate the profiles of leprosy cases in the study population. Results: A total of 110 patients were included in the study, with a mean age of 33.54 years. The majority of the patients were Nepalese. The mean duration of the disease was 17.6 months, with a wide range observed. Borderline leprosy was the most common clinical presentation. Bacteriological positivity was observed in 85% of the cases, indicating active transmission of the disease within the community. A significant proportion of the patients (97%) belonged to the low to middle-class category. A reaction was seen in 14.5% of the patients. Conclusion: Despite efforts toward its elimination, leprosy remains a public health problem across borders. The high burden of leprosy cases, coupled with socioeconomic challenges and active transmission, underscores the need for continued vigilance and targeted interventions. Addressing cross-border dynamics, enhancing access to healthcare services, and implementing comprehensive control strategies are essential for effectively controlling leprosy. Key words: Leprosy, Clinical, Epidemiological, Cross-sectional study, Nepal–India border, Post-elimination era

Immunomodulators in leprosy: A narrative review

Background: Leprosy is a chronic infectious disease caused by Mycobacterium leprae. Current therapeutic regimen, like the multidrug therapy (MDT), are effective in treating most cases, but new cases continue to emerge in Indonesia every year. While multidrug therapy alone is adequate for treating leprosy, there is a need for adjuvant treatment options to boost the host’s immune system to prevent the worsening of leprosy and reduce the activation of M. leprae, such as immunomodulators. Discussion: Immunomodulators are drugs that can stimulate the body’s natural and adaptive defense mechanisms, acting as either immunosuppressants or immunostimulants. To understand how immunomodulatory drugs (IMiDs) work, it is important to understand the role of immunity. This article reviews the role of immunity in leprosy and discusses various immunomodulators that have been developed or investigated to enhance the host’s immune system. Substances like levamisole, thalidomide, zinc, selenium, as well as vitamins A, D, E, and C have been clinically tried in various combinations and durations, showing promise as immunomodulating agents. Conclusion: Studies have suggested that immunomodulating agents may be considered as adjuncts to MDT to enhance the elimination and clearance of bacteria, making them potential recommendations for leprosy treatment.

Employing advanced computational drug discovery techniques to identify novel inhibitors against ML2640c protein: a potential therapeutic approach for combatting leprosy.

Leprosy, caused by Mycobacterium leprae, remains a significant global health challenge, necessitating innovative approaches to therapeutic intervention. This study employs advanced computational drug discovery techniques to identify potential inhibitors against the ML2640c protein, a key factor in the bacterium's ability to infect and persist within host cells. Utilizing a comprehensive methodology, including virtual screening, re-docking, molecular dynamics simulations, and free energy calculations, we screened a library of compounds for their interaction with ML2640c. Four compounds (24349836, 26616083, 26648979, and 26651264) demonstrated promising inhibitory potential, each exhibiting unique binding energies and interaction patterns that suggest a strong likelihood of disrupting the protein function. The study highlights the efficacy of computational methods in identifying potential therapeutic candidates, presenting compound 26616083 as a notably potent inhibitor due to its excellent binding affinity and stability. Our findings offer a foundation for future experimental validation and optimization, marking a significant step forward in the development of new treatments for leprosy. This research not only advances the fight against leprosy but also showcases the broader applicability of computational drug discovery in tackling infectious diseases.

Human Skin as an Ex Vivo Model for Maintaining Mycobacterium leprae and Leprosy Studies.

The in vitro cultivation of M. leprae has not been possible since it was described as causing leprosy, and the limitation of animal models for clinical aspects makes studies on leprosy and bacteria-human host interaction a challenge. Our aim was to standardize the ex vivo skin model (hOSEC) to maintenance and study of M. leprae as an alternative animal model. Bacillary suspensions were inoculated into human skin explants and sustained in DMEM medium for 60 days. Explants were evaluated by RT-PCR-16SrRNA and cytokine gene expression. The viability and infectivity of bacilli recovered from explants (D28 and D60) were evaluated using the Shepard's model. All explants were RT-PCR-16SrRNA positive. The viability and infectivity of recovered bacilli from explants, analyzed after 5 months of inoculation in mice, showed an average positivity of 31%, with the highest positivity in the D28 groups (80%). Furthermore, our work showed different patterns in cytokine gene expression (TGF-β, IL-10, IL-8, and TNF-α) in the presence of alive or dead bacilli. Although changes can be made to improve future experiments, our results have demonstrated that it is possible to use the hOSEC to maintain M. leprae for 60 days, interacting with the host system, an important step in the development of experimental models for studies on the biology of the bacillus, its interactions, and drug susceptibility.

Any resurgence of leprosy cases in the Togo’s post-elimination period? Trend analysis of reported leprosy cases from 2010 to 2022

BackgroundLeprosy, or Hansen’s disease, is a chronic infectious disease caused by Mycobacterium leprae. Togo achieved the target of eliminating leprosy as a public health problem in 2000 (less than 1 case/10 000 population). However, new cases of leprosy are still being reported. The aim of this study was to describe and map trends of leprosy cases notified in Togo from 2010 to 2022.MethodsThis was a descriptive cross-sectional study covering a thirteen-year period from January 1, 2010, to December 31, 2022. The data of the study were leprosy surveillance system’s data collected monthly between 2010 and 2022. The estimated number of leprosy cases and the incidence rate of leprosy cases were reported for the whole population by region, by district, by calendar year (2010–2022) and by target sub-population (children under 15, women and people with disabilities). Observed case incidence rates were mapped by health district and by year.ResultsFrom January 1, 2010, to December 31, 2022, 1031 new cases of leprosy were diagnosed in Togo. The median age of subjects was 46 years (interquartile range: 33–60), with extremes from 4 to 96 years. Half the subjects were women (50.7%). Variations in the leprosy incidence rate by year show an increase between 2010 and 2022, from 0.7 cases /100,000 population to 1.1 /100,000 population respectively. From 2010 to 2022, the proportion of cases in children remained low, between 0 and 9%. The proportion of women fluctuated between 39.7% and 67.2% between 2010 and 2017, then stabilized at an average of 50% between 2018 and 2022. The proportion of multi-bacillary leprosy cases increased quasi-linearly between 2010 and 2022, from 70 to 96.6%. Mapping of leprosy cases showed that leprosy was notified in all Togo health districts during the study period, apart from the Lacs district, which reported no leprosy cases.ConclusionTogo has achieved the elimination of leprosy as a public health problem. However, the increase in the number of new leprosy cases and the proportion of leprosy cases in children indicate that transmission of the disease is continuing and that supplementary measures are needed.

The Presence of esat-6 and cfp10 and Other Gene Orthologs of the RD 1 Region in Non-Tuberculous Mycobacteria, Mycolicibacteria, Mycobacteroides and Mycolicibacter as Possible Impediments for the Diagnosis of (Animal) Tuberculosis.

The Esx-1 family proteins of the Type VII secretion systems of Mycobacterium bovis and Mycobacterium tuberculosis have been assessed and are frequently used as candidates for tuberculosis (TB) diagnosis in both humans and animals. The presence of ESAT-6 and CFP 10 proteins, which are the most immunogenic proteins of the Esx-1 system and have been widely investigated for the immunodiagnosis of tuberculosis, in some Mycobacteriaceae and in Mycobacterium leprae, poses limitations for their use in specific diagnoses of TB. As such, to improve the specificity of the ESAT-6/CFP 10-based cell-mediated immunity (CMI) assays, other proteins encoded by genes within and outside the RD 1 region of the esx-1 locus have been evaluated as candidate antigens for CMI, as well as to investigate humoral responses in combination with ESAT-6 and or CFP 10, with varying specificity and sensitivity results. Hence, in this study, we evaluated various non-tuberculous mycobacteria (NTM), Mycolicibacterium, Mycolicibacter and Mycobacteroides species genomes available on the NCBI database for the presence and composition of the RD1 region of the esx-1 locus. In addition, we also assayed by polymerase chain reaction (PCR) and sequencing of Mycobacteriaceae available in our culture collection for the presence and sequence diversity of esxA and esxB genes encoding ESAT-6 and CFP 10, respectively. Whole genome sequence (WGS) data analysis revealed the presence of RD 1 gene orthologs in 70 of the over 100 published genomes of pathogenic and non- pathogenic Mycobcteriaceae other than tuberculosis. Among species evaluated from our culture collection, in addition to earlier reports of the presence of esxA and esxB in certain Mycolicibacterium, Mycolicibacterium septicum/peregrinum, Mycolicibacterium porcinum and Mycobacterium sp. N845T were also found to harbour orthologs of both genes. Orthologs of esxA only were detected in Mycobacterium brasiliensis, Mycolicibacterium elephantis and Mycolicibacterium flouroantheinivorans, whereas in Mycolicibacter engbackii, Mycolicibacterium mageritense and Mycobacterium paraffinicum, only esxB orthologs were detected. A phylogenetic analysis based on esxA and esxB sequences separated slow-growing from rapidly growing bacteria. These findings strengthen previous suggestions that esxA and esxB may be encoded in the majority of Mycobacteriaceae. The role of the Esx-1 system in both pathogenic and non-pathogenic Mycobacteriaceae needs further investigation, as these species may pose limitations to immunological assays for TB.

Moran’s Index Spatial Analysis On The New Case Detection Rate of Leprosy in West Java 2022

Introduction: Leprosy is a chronic infectious disease, neglected tropical disease caused by Mycobacterium leprae. Leprosy is divided into two types based on the number of lesions on the skin. If there are less than 5 lesions, leprosy is classified as paucibacillary (PB) and if there are more than 5 lesions, leprosy is classified as multibacillary (MB). Indonesia is ranked third in the world after Brazil with 762 new cases of grade 2 disability leprosy by 2022. Therefore, spatial analysis of the new case detection rate of leprosy in West Java 2022 is needed. Objective: To determine wheter there are spatial dependency with the new case detection rate of leprosy in West Java 2022 Method: Quantitative research that utilises secondary data by conducting spatial analysis on the new case detection rate of leprosy in West Java 2022 using Moran's Index and LISA tests. Result: There are 5 areas namely Bekasi, Karawang, Subang, Indramayu, and Cirebon that have not yet reached the national target in eliminating leprosy. The value of Moran's I = 0.241 and p-value 0.0090 < ? (0.05) which means that there is a weak positive spatial dependence on the number of new leprosy cases in West Java Province 2022. There is 1 region that is in the high-high quadrant, namely Cirebon and there are 4 regions in the low-low quadrant, namely Cimahi City, Bandung City, Garut, and Tasikmalaya. Conclusion: A small number of areas in West Java Province still have not reached the national target of eliminating leprosy, which is a CDR of <5 per 100.000 population, namely Bekasi, Karawang, Subang, Indramayu, and Cirebon. There are 5 areas that have spatial linkages in the new case detection rate of leprosy in West Java Province in 2022 based on the significance value. The region in the high-high quadrant is Cirebon and there are 4 regions in the low-low quadrant, namely Cimahi City, Bandung City, Garut District, and Tasikmalaya District.

MIP vaccine in leprosy: A scoping review and future horizons.

Mycobacterium Indicus Pranii (MIP) vaccine is a killed vaccine developed in India for leprosy with immunotherapeutic as well as immunoprophylactic effects. MIP, earlier known as Mycobacterium welchii, is a rapidly growing non-pathogenic mycobacterium. The novelty of this bacterium is due to its translational application as an immunotherapeutic agent. When administered intradermally, the vaccine induces cell-mediated immunity in the host towards Mycobacterium leprae. It leads to faster clinical and histopathological improvement, rapid bacillary clearance, and also lepromin conversion in anergic leprosy patients. The beneficial role of the MIP vaccine in augmenting the therapeutic efficacy of Multidrug Therapy (MDT), particularly in highly bacillated leprosy patients, is well documented in various studies from India. The role of the vaccine in reactional states is controversial, with varied results in different studies. Overall, it is found to decrease the frequency of type 2 lepra reactions and is useful in recalcitrant erythema nodosum leprosum. Even though there may be an increased likelihood of type 1 reactions, no additional nerve function impairment is attributed to the vaccine in various studies. In household contacts of leprosy who are administered MIP, it is noted to confer protection from disease lasting up to 10 years. It may prove to be a cost-effective strategy in national leprosy programmes. Apart from local injection site reactions, the vaccine is relatively safe, but it is not recommended in pregnancy and lactation. This article provides an overview of the MIP vaccine's clinical application in the context of leprosy spanning over 40 years. It also considers the vaccine's possible future applications in the management of disease-related complications and achieving the long-term goal of zero leprosy.

A Review on Treatment of Leprosy by using Medicinal Plants

Traditionally plants have been used as medicinal agents for treatment of disease such as antihypertensive, antitussive, diuretics, antidiabetics, Antileprotics etc. The impressive number of allopathic drugs has been isolated from the natural sources. Leprosy is a disease, caused by the bacteria Mycobacterium leprae, which causes damage to the skin. Many of plants are used for treatment of leprosy.

Leprosy: Comprehensive insights into pathology, immunology, and cutting-edge treatment strategies, integrating nanoparticles and ethnomedicinal plants.

Mycobacterium leprae is the causative agent responsible for the chronic disease known as leprosy. This condition is characterized by dermal involvement, often leading to peripheral nerve damage, sensory-motor loss, and related abnormalities. Both innate and acquired immunological responses play a role in the disease, and even in individuals with lepromatous leprosy, there can be a transient increase in T cell immunity during lepromatous reactions. Diagnosing of early-stage leprosy poses significant challenges. In this context, nanoparticles have emerged as a promising avenue for addressing various crucial issues related to leprosy. These include combatting drug resistance, mitigating adverse effects of conventional medications, and enhancing targeted drug delivery. This review serves as a comprehensive compilation, encompassing aspects of pathology, immunology, and adverse effects of multidrug delivery systems in the context of leprosy treatment. Furthermore, the review underscores the significance of ethnomedicinal plants, bioactive secondary metabolites, and nanotherapeutics in the management of leprosy. It emphasizes the potential to bridge the gap between existing literature and ongoing research efforts, with a profound scope for validating traditional claims, developing herbal medicines, and formulating nanoscale drug delivery systems that are safe, effective, and widely accepted.

Multibacillary lepromatous leprosy detected in Olmedo, Ecuador. Case report

Introduction: hansen's disease or leprosy is an infectious disease caused by Mycobacterium leprae, which mainly affects the skin, peripheral nerves and mucosa of the upper respiratory tract. It has a chronic course as old as humanity itself.Clinical case: the clinical case of a 51-year-old male patient, native and from the Olmedo canton, province of Manabí, with a farmer occupation, who began to present in November 2008 with macular, hyperchromic skin lesions, which progressed, is presented. to multiple nodules, located in the lower limbs and then spreading to the upper limbs, thorax and abdomen, neck and face. The set of clinical parameters and positive biopsy results allowed the case to be classified as Lepromatous Leprosy (multibacillary). Triple multitherapy (Dapsone, Rifampicin, Clofazimine) was prescribed, showing an improvement in the number and size of the lesions at the end of therapy.Discussion: this is the first clinical case of lepromatous leprosy detected in the Olmedo canton, Manabí province, Ecuador. Due to the novelty in the territory, the patient had this condition for years without being diagnosed or treated adequately and even once under treatment, there were delays in acquiring the medications that extended their application from 12 months to a period of 25 months.Conclusions: the clinical case presented highlights that Hansen's Disease is an emerging health problem in the Olmedo canton, highlighting the need to make sustained efforts for its identification, treatment and prevention in the territory

Leprosy in Post-elimination Era: A Study Conducted in Rural Tertiary Settings in North India.

Background Leprosy is an age-old disease caused by Mycobacterium leprae. The disease was declared eradicated in India in 2005. Many new cases are still being identified in the outdoor patient department. This study was undertaken to understand the epidemiological, clinical, and social aspects of leprosy among new patients, and assessthe current situation regarding caseload and presentation. Material and methods This study was designed as an observational study. It was carried out in people newly diagnosed with leprosyattending the outpatient department of Dermatology, Venereology, and Leprology in the tertiary care hospital in Uttar Pradesh University of Medical Sciences from July 2022 to January 2024. A total of 231 people afflicted with leprosy were included in the study. The data collected was statistically analyzed to identify demographic and social patterns, clinical presentations, and features associated with leprosy. Result Out of these 231 patients, 139 (60.17%) were male and 92 (39.83%) were female. Most cases belonged to the age group 40-59 years 87 (37.66%). History of close contact with an afflicted person was present in 34 (14.71%). Clinically, most patients belong to the borderline tuberculoid (BT) type. Only 24 (10.4%) patients were found positive for M. leprae by slit-skin smear examination. The ulnar nerve was the most common nerve involved in 63 (27.27%) cases. Trophic ulcers were the predominant deformity in 34 (14.7%), followed by foot drop in 13 (5.62 %). Conclusion The present study provides an overview of the prevailing trends of Leprosy within a specific region in the post-elimination era. The findings underscore the significance of the ongoing National Leprosy Eradication Program (NLEP) program and stress the importance of aligning them with the common goal of eliminating the burden and stigma of Leprosy from society.

Ancient Mycobacterium leprae genome reveals medieval English red squirrels as animal leprosy host

Leprosy, one of the oldest recorded diseases in human history, remains prevalent in Asia, Africa, and South America, with over 200,000 cases every year.1,2 Although ancient DNA (aDNA) approaches on the major causative agent, Mycobacterium leprae, have elucidated the disease's evolutionary history,3,4,5 the role of animal hosts and interspecies transmission in the past remains unexplored. Research has uncovered relationships between medieval strains isolated from archaeological human remains and modern animal hosts such as the red squirrel in England.6,7 However, the time frame, distribution, and direction of transmissions remains unknown. Here, we studied 25 human and 12 squirrel samples from two archaeological sites in Winchester, a medieval English city well known for its leprosarium and connections to the fur trade. We reconstructed four medieval M.leprae genomes, including one from a red squirrel, at a 2.2-fold average coverage. Our analysis revealed a phylogenetic placement of all strains on branch 3 as well as a close relationship between the squirrel strain and one newly reconstructed medieval human strain. In particular, the medieval squirrel strain is more closely related to some medieval human strains from Winchester than to modern red squirrel strains from England, indicating a yet-undetected circulation of M.leprae in non-human hosts in the Middle Ages. Our study represents the first One Health approach for M.leprae in archaeology, which is centered around a medieval animal host strain, and highlights the future capability of such approaches to understand the disease's zoonotic past and current potential.

PEMBERDAYAAN PENDERITA KUSTA DENGAN KECACATAN DERAJAT 2 DAN KELUARGANYA DALAM REHABILITASI DAN PERAWATAN DIRI SECARA MANDIRI

Leprosy is a chronic infectious disease caused by Mycobacterium leprae. Leprosy is the main cause of permanent physical disability. Defects in the hands and feet are divided into three levels of disability according to WHO (1988), level 0 (no anesthesia and anatomical abnormalities), level 1 (anesthesia, no anatomical abnormalities) and level 2 (anatomical abnormalities). There is no effective therapy for leprosy disability, so holistic therapy such as medical rehabilitation is needed. The Community Partnership Program (PKM) partner is a leprosy sufferer, a 63 years old woman with grade 2 disability and her family. The patient also experienced paralysis in both legs due to a stroke which was a complication of the diabetes mellitus. The method for implementing PKM activities is by providing training on self-rehabilitation and self-care so that complications due to leprosy disability can prevented, and providing wheelchairs to help partners mobilize. Indicators of achievement include moist extremity skin, no new wounds appearing, contractures on the patient's hands and both of legs not getting worse and the patient getting a wheelchair has reached 100%.

Whole blood transcriptomics reveals the enrichment of neutrophil activation pathways during erythema nodosum leprosum reaction.

Patients with the multibacillary form of leprosy can develop reactional episodes of acute inflammation, known as erythema nodosum leprosum (ENL), which are characterized by the appearance of painful cutaneous nodules and systemic symptoms. Neutrophils have been recognized to play a role in the pathogenesis of ENL, and recent global transcriptomic analysis revealed neutrophil-related processes as a signature of ENL skin lesions. In this study, we expanded this analysis to the blood compartment, comparing whole blood transcriptomics of patients with non-reactional lepromatous leprosy at diagnosis (LL, n=7) and patients with ENL before administration of anti-reactional treatment (ENL, n=15). Furthermore, a follow-up study was performed with patients experiencing an ENL episode at the time of diagnosis and after 7 days of thalidomide treatment (THAL, n=10). Validation in an independent cohort (ENL=8; LL=7) was performed by RT-qPCR. An enrichment of neutrophil activation and degranulation-related genes was observed in the ENL group, with the gene for the neutrophil activation marker CD177 being the most enriched gene of ENL episode when compared to its expression in the LL group. A more pro-inflammatory transcriptome was also observed, with increased expression of genes related to innate immunity. Validation in an independent cohort indicated that S100A8 expression could discriminate ENL from LL. Supernatants of blood cells stimulated in vitro with Mycobacterium leprae sonicate showed higher levels of CD177 compared to the level of untreated cells, indicating that the leprosy bacillus can activate neutrophils expressing CD177. Of note, suggestive higher CD177 protein levels were found in the sera of patients with severe/moderate ENL episodes when compared with patients with mild episodes and LL patients, highlighting CD177 as a potential systemic marker of ENL severity that deserves future confirmation. Furthermore, a follow-up study was performed with patients at the time of ENL diagnosis and after 7 days of thalidomide treatment (THAL, n=10). Enrichment of neutrophil pathways was sustained in the transcriptomic profile of patients undergoing treatment; however, important immune targets that might be relevant to the effect of thalidomide at a systemic level, particularly NLRP6 and IL5RA, were revealed. In conclusion, our study reinforces the key role played by neutrophils in ENL pathogenesis and shed lights on potential diagnostic candidates and novel therapeutic targets that could benefit patients with leprosy.

A comparative study on the detection of Mycobacterium leprae DNA in urine samples of leprosy patients using Rlep-PCR with other conventional samples.

Mycobacterium leprae causes leprosy that is highly stigmatized and chronic infectious skin disease. Only some diagnostic tools are being used for the identification M. leprae in clinical samples, such as bacillary detection, and histopathological tests. These methods are invasive and often have low sensitivity. Currently, the PCR technique has been used as an effective tool fordetecting M. leprae DNA across different clinical samples. The current study aims to detect M. leprae DNA in urine samples of untreated and treated leprosy patients using the Rlep gene (129 bp) and compared the detection among Ridley-Jopling Classification. Clinical samples (Blood, Urine, and Slit Skin Smears (SSS)) were collected from leprosy and Non-leprosy patients. DNA extraction was performed using standard laboratory protocol and Conventional PCR was carried out for all samples using Rlep gene target and the amplicons of urine samples were sequenced by Sanger sequencing to confirm the Rlep gene target. The M. leprae DNA was successfully detected in all clinical samples across all types of leprosy among all the study groups using RLEP-PCR. Rlep gene target was able to detect the presence of M. leprae DNA in 79.17% of urine, 58.33% of blood, and 50% of SSS samples of untreated Smear-Negative leprosy patients. The statistical significant difference (p = 0.004) was observed between BI Negative (Slit Skin Smear test) and RLEP PCR positivity in urine samples of untreated leprosy group. The PCR positivity using Rlep gene target (129 bp) was highest in all clinical samples among the study groups, across all types of leprosy. Untreated tuberculoid and PNL leprosy patients showed the highest PCR positivity in urine samples, indicating its potential as a non-invasive diagnostic tool for leprosy and even for contact screening.