Erythema Nodosum Leprosum Reaction Research Articles

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.

Single-nucleotide polymorphism of interleukin-10 promoter (IL-10 -819C/T) in leprosy patients with and without erythema nodosum leprosum, and healthy household contacts.

Leprosy, caused by Mycobacterium leprae, is a chronic infectious disease that impacts the skin and peripheral nerves, causing long-term disability. The invasion of M. leprae into the body triggers immunologic responses and single single-nucleotide polymorphisms in cytokine-encoding genes may influence predisposition and susceptibility, possibly predicting the incidence of leprosy reactions. The aim of this study was to assess the gene polymorphism of interleukin-10 promoter IL-10 -819C/T in leprosy patients, leprosy patients with erythema nodosum leprosum (ENL) reaction, and household contacts. A total of 54 individuals were included, with 18 in each group. Skin smear and histopathologic examinations were used to confirm the diagnosis of leprosy and ENL. The polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) technique was used to determine the polymorphism. The results confirmed the presence of polymorphism of which all TT, CT, and CC genotypes presented. The TT genotype was most prevalent in household contacts (94.4%) followed by ENL (50%), and leprosy patients (44.4%). The CT genotype was most frequently detected in leprosy patients (50%), followed by ENL cases (44.4%), and household contacts (5.56%). In contrast, CC was mostly presented in ENL cases (5.56%), only 1% in leprosy patients, and absent among household contacts. Although the most prevalent allele in all three groups was the T allele, the C allele presented in 27% and 30% of ENL and leprosy patients, respectively and only 5% in household contact individuals. This study suggests that the polymorphism variations of IL-10 -819C/T are higher in leprosy and ENL patients compared to household contacts. Since this data is preliminary, larger studies are needed.

Analysis of interleukin 7 and platelet-derived growth factor-BB mRNA expression as potential markers in erythema nodosum leprosum.

Erythema nodosum leprosum (ENL) is an immunological complication of leprosy characterized by acute inflammation of the skin, nerves, and other organs. Identifying laboratory parameters is important for early diagnosis of leprosy reactions. Various cytokine biomarkers have been examined and only a few studies have reported on angiogenesis in leprosy. This study aims to understand the pathomechanism of ENL by examining IL-7 and platelet-derived growth factor (PDGF)-BB mRNA expression that can be the development and consideration of new effective therapies to prevent reactions, recurrences, and defects in leprosy. The study used a cross-sectional analytic design. Sampling was done by peripheral blood from the patient and measuring mRNA expression with specific primers RT-PCR. The expression of mRNA IL-7 and PDGF-BB was significantly different between multibasilar patients without reaction and with ENL reaction, where there was an increased expression in ENL patients. This could be used as the development of potential biomarkers in ENL and development of new therapeutic intervention pathways in ENL.

Systemic manifestation of necrotic erythema nodosum leprosum: A case report of a fatal leprosy.

Necrotic erythema nodosum leprosum (ENL) is an uncommon manifestation of type 2 lepra reaction, encountered in lepromatous and borderline lepromatous cases of leprosy. Necrotic ENL is associated with the involvement of multiple organs, therefore delayed diagnosis and treatment will lead to complications and poor prognosis. The aim of this case report was to report a challenging case of necrotic ENL misdiagnosed with multiple cellulitis since there were no signs of prior leprosy nor had any antimycobacterial treatment. A 45-year-old man was presented to the surgery department of Dr. Zainoel Abidin Hospital, Banda Aceh, Indonesia, with complaints of fever, joint pain, and painful tender skin lesions with ulceration over the trunk, extremities, and ears for one month. The patient was diagnosed clinically with multiple cellulitis and underwent a debridement procedure. Clinical improvement was absent, the patient was then consulted to the dermatology department. Physical examination showed normal vital signs, madarosis, inguinal lymphadenopathy, thickening of nerves, and sensation of numbness in both hands and feet. Laboratory examinations on admission showed leucocytosis, anemia, thrombocytopenia, hypoalbuminemia, hypocalcemia, and elevated creatinine and ureum level. A slit skin smears examination yielded positive acid-fast bacilli (AFB) with a bacteriological index (BI) value of 3+ and morphological index (MI) of 72%. The patient was diagnosed with lepromatous leprosy with necrotic ENL reaction. Intravenous methylprednisolone and cefoperazone-sulbactam were given. Multidrug therapy mulitbacillary (MDT-MB) without dapsone, and ofloxacin 400 mg was initiated. On day 17, the patient had septic shock. The patient became unconscious and experienced death. This case highlights that medical professionals should be aware of the various manifestations of necrotic ENL to correctly diagnose and provide treatment as soon as possible to prevent mortality, especially in leprosy-endemic country, Indonesia.

Borderline Lepromatous Leprosy with Severe Erythema Nodosum Leprosum: A Case Report

Background: Leprosy is a chronic granulomatous infectious disease caused by Mycobacterium leprae (M. leprae) that primarily infects Schwann cells in the peripheral nerves, leading to nerve damage and the development of disabilities. In 2018, Indonesia was the third country with the most leprosy cases in the world. Erythema nodosum leprosum (ENL), also known as type II leprosy reaction, is a severe immune-mediated complication of multibacillary leprosy. Purpose: To report a case of borderline lepromatous leprosy with severe ENL. Case: A 49-year-old Balinese man presented with multiple tender erythematous skin nodules all over his body, fever, arthralgia, bilateral cervical lymphadenopathy, and sensory loss for the past week. The acid-fast bacilli bacteriological examination showed a positive result. The patient was diagnosed with borderline lepromatous (BL) leprosy with severe ENL and was treated with multibacillary multidrug therapy (MB MDT), methylprednisolone, and other symptomatic medications. After 1 month of treatment, there was an improvement in skin lesions. The MB-MDT treatment was continued and methylprednisolone was planned to be tapered down gradually. Discussion: Approximately 20-50% of all leprosy patients show leprosy reactions in the course of the disease. The goals of treatment for severe ENL are to control inflammation, reduce pain, treat neuritis to prevent nerve dysfunction and contractures, and prevent recurring ENL. The prognosis of leprosy with ENL reactions depends on the severity of the occurring leprosy reaction; early diagnosis and prompt treatment; and patient compliance with treatment. Conclusion: Early diagnosis and treatment are essential to avoid deformities in leprosy patients.

Comparison between oral pentoxifylline + corticosteroid and oral corticosteroid alone for severe erythema nodosum leprosum

Background: Severe erythema nodosum leprosum (ENL) is common but difficult to treat. Long term use of systemic corticosteroid causes side effects. This study compares the use of a combination of pentoxifylline + oral corticosteroids with a single oral corticosteroid in leprosy patients with severe ENL reactions. Parameters measured include skin RSA score, systemic RSA, total corticosteroid dose, resolution time of skin lesions, improvement of pain VAS and treatment side effects. Methods: 29 subjects with severe ENL reactions are allocated randomly into two groups which receive oral pentoxifylline + methylprednisolone, and oral placebo + methylprednisolone for 12 weeks. The starting dose of pentoxifylline are 400 mg thrice daily for 4 weeks, tapered to 400 mg daily every 4 weeks. Methylprednisolone is based on WHO guideline. Results: In the end, the median of cutaneous RSA score in pentoxifylline vs placebo group is 4 (0-5) vs 3 (0- 5). The median of systemic RSA score in pentoxifylline vs placebo group is 0 (0-6) vs 0 (0-5). The median of total corticosteroid doses in pentoxifylline vs placebo group is 156 (120-200) mg vs 136 (96-200) mg. The median of resolution time in pentoxifylline vs placebo group is 6 (0-12) weeks vs 6 (0-12) weeks. The median of change of pain VAS score in pentoxifylline vs placebo group is 5 (0-6) vs 3 (-3-6). No statistically significant difference (p>0,05) are found in all parameters, including side effects. Conclusion: Combination of oral pentoxifylline + corticosteroid is not proven to be more effective. Both are safe. Keywords: combination, corticosteroid, erythema nodosum leprosum, pentoxifylline

A Rarely Employed Therapeutic Pearl of Split-Dose Oral Corticosteroid in Severe Type 2 or Erythema Nodosum Leprosum Reaction in Lepromatous Leprosy and its Therapeutic Rationale.

A Rarely Employed Therapeutic Pearl of Split-Dose Oral Corticosteroid in Severe Type 2 or Erythema Nodosum Leprosum Reaction in Lepromatous Leprosy and its Therapeutic Rationale.

Immunopathogenesis of erythema nodosum leprosum

Leprosy is a chronic granulomatous disease. Its etiology is caused by Mycobacterium leprae (M. leprae). This disease may affect the skin, however it also may affect peripheral nervous system and other organs. The immunologic mechanism of erythema nodosum leprosum (ENL) has not been fully understood. The ENL reaction is the reaction of deposition in immune complex in the blood vessels. Recent studies have reported the association of several immunological markers with leprosy reactions, but inconsistent results have been obtained. Neutrophils are considered a histologic feature of ENL, histologic examination of ENL skin lesions revealed a strong perivascular neutrophil infiltrate throughout the dermis and subcutis. Another thing to note is T lymphocytes. It has been showed that the T-cell counts in peripheral blood of ENL patients is higher. Other immunological processes that play a role in EL immunopathogenesis involve T-helper (Th) cells, T regulatory cells, B cells, and other cytokines.

Reversal reaction with nodules that initially diagnosed as erythema nodosum leprosum in borderline lepromatous leprosy

Background:  The reversal leprosy reaction generally manifest as larger, swollen, red and shiny the pre-existing skin lesions which are accompanied with pain. This reaction can be manifest as infiltration and nodules which resembles the features of erythema nodosum leprosum (ENL)  leprosy  reaction.  Case Illustration:  We present a case of 26-year-old woman with erythematous nodules on almost all the body since two weeks  ago. She had  pre-existing erythema plaque on the right elbow became more erythematous and arouse with partially felt pain, fever, and malaise. Nodule lesions in BL leprosy with reaction need to be distinguished between ENL and reversal reaction  because  they may affect therapy and prognosis.   Discussion:  T he patient was diagnosed as BL leprosy with ENL reaction e rythematous nodules on the face, both upper and lower arms, upper and lower limbs, knees, that   partially  felt  pain , but after more careful history and histopathologic examination of the lesions, the nodules didn’t match to the histopathologic features of ENL reaction. Correlation between the clinical and histopathological findings in the form of acid fast bacilli (AFB) and  granulomatous inflammation grenz zone with epithelioid cells and lymphocyte cells infiltrations in the dermis  established the diagnosis of BL leprosy with reversal reaction .  Conclusion:  Erythema nodule lesions in leprosy can be an ENL or reversal reaction or a leprosy lesion in type BL leprosy.   Reversal reaction  should always be considered when diagnosing a skin-colored nodule in leprosy.  Appropriate clinical and histopathological findings of the  skin-colored  nodules are needed  to establish the definite diagnosis of reversal reaction.   Keywords:  leprosy, borderline lepromatous, n odule, reversal reaction, erythema nodosum leprosum

Palatal perforation in Erythema Nodosum Leprosum (ENL) case of hansen’s disease: A case report

Leprosy (Hansen’s disease) is mainly affecting skin & nerve. Lepra reaction is common amongst leprosy patient. We report 64 year female patient who completed MB-MDT treatment and was on steroid therapy for treatment of ENL reaction. We observed unusual presentation of palatal perforation which rarely seen in ENL case. On examination it found that perforation was present on hard palate midline at the level of first molar tooth. Such type of presentation rarely seen in ENL cases till date few such reports have published in the journals. Keyword: Multi-bacillary leprosy case, Chronic ENL reaction, palatal perforation.

Lepra reactions in new leprosy cases at diagnosis: A study of 50 Pakistani patients.

To determine the occurrence and characteristics of the two types of lepra reactions in new leprosy cases at initial diagnosis. The retrospective descriptive study was conducted at the Marie Adelaide Leprosy Centre, Karachi, and comprised all new leprosy cases registered from January 1, 2016, to June 30, 2018. Data was collected from the medical record database using a predesigned proforma. Of the 50 cases, 2(4%) were children and 48 (96%) were adults, with overall age ranging from 12 to 85 years. There were 41(82%) males and 9(18%) females.. Of the total, 30(60%) cases presented with type 1 reaction and 20(40%) with type 2. Further, 30(60%) cases were classified as borderline lepromatous. Among them, 17(57%) had type 2 reaction. Inflamed plaques were the main feature in 27(90%) cases of type 1. Crops of painful, erythematous nodules were seen in 19(95%) cases of type 2. Lepra reactions were found to be a presenting feature in a significant number of new leprosy cases at initial diagnosis.

γδ T cells are associated with inflammation and immunopathogenesis of leprosy reactions

BackgroundLeprosy reactions appear episodically in leprosy patients, which lead to high inflammation, morbidity and peripheral nerve damage. The role of Th17 cell has been well studied in leprosy reactions but the role of γδ or unconventional T cells which is an other major source of IL-17 in many diseases, not studied in leprosy reactional episodes. ObjectiveThe aim of the present study to elucidate the role of γδ T cells in leprosy reactions. MethodologyA total of 40 untreated non-reaction and reactions patients were recruited. PBMCs were isolated and stimulated with M. leprae sonicated antigen (MLSA) for 48 h and immuno-phenotyping was done using flow cytometry. Moreover, γδ T cells were isolated by Magnetic beads technology and mRNA expression of IL-17, IFN-γ, TGF-β and FOXP3 were analyzed by real-time PCR (qPCR) and cytokine was estimated in the culture supernatant by ELISA. Resultsγδ T cells were significantly increased in both Reversal reaction (RR) and Erythema nodosum leprosum (ENL) reaction patients. These cells produced significant amount of IL-17 and IFN-γ. Furthermore, CD3+TCRγδ+ T cells expressed transient FOXP3 with a low amount of TGF-β in both reactions as compared to stable patients. Moreover, low TGF-β producing TCR-γδ cells were associated with low phosphorylation of STAT5A. ConclusionThis study will add to our understanding of the immunological features that mediate and regulate the pathogenesis of leprosy and may helpful to reduce the immuno-pathogenesis of leprosy reaction by targeting these cells.

Complement C1q expression in Erythema nodosum leprosum.

Complement C1q is a soluble protein capable of initiating components of the classical pathway in host defence system. In earlier qualitative studies, C1q has been implicated in the pathogenesis of Erythema Nodosum Leprosum (ENL). However, little is known about the role of this complement in ENL reaction. In the present study we described the protein level of C1q production and its gene expression in the peripheral blood and skin biopsies in patients with ENL reaction and lepromatous leprosy (LL) patient controls before and after treatment. Thirty untreated patients with ENL reaction and 30 non-reactional LL patient controls were recruited at ALERT Hospital, Ethiopia. Peripheral blood and skin biopsies were obtained from each patient before and after treatment. The level of circulating C1q in the plasma was determined by enzyme-linked immunosorbent assay. The mRNA expression of the three C1q components, C1qA, C1qB, and C1qC in the peripheral blood and skin biopsies was determined by qPCR. Circulating C1q in the peripheral blood of untreated ENL patients was significantly decreased compared to LL patient controls. Untreated ENL patients had increased C1q gene expression in the peripheral blood compared to LL controls. Similarly, C1qA and C1qC gene expression were substantially increased in the skin biopsies of untreated ENL patients compared to LL controls. However, after treatment none of these genes show significant difference in both groups. In conclusion, while circulating C1q is inversely correlated with active ENL reactions, its gene expression is directly correlated with ENL. The decreased circulating C1q may suggest the utilization of C1q in immune-complex formation in these patients. Therefore, C1q could be a potential diagnostic marker for active ENL reactions as well as for monitoring ENL treatment.

The Effects of Prednisolone Treatment on Cytokine Expression in Patients with Erythema Nodosum Leprosum Reactions.

Erythema nodosum leprosum (ENL) is a systemic inflammatory complication occurring mainly in patients with lepromatous leprosy (LL) and borderline lepromatous leprosy. Prednisolone is widely used for treatment of ENL reactions but clinical improvement varies. However, there is little good in vivo data as to the effect of prednisolone treatment on the pro-inflammatory cytokines in patients with ENL reactions. As a result, treatment and management of reactional and post-reactional episodes of ENL often pose a therapeutic challenge. We investigated the effect of prednisolone treatment on the inflammatory cytokines TNF, IFN-γ, IL-1β, IL-6, and IL-17 and the regulatory cytokines IL-10 and TGF-β in the skin lesion and blood of patients with ENL and compared with non-reactional LL patient controls. A case–control study was employed to recruit 30 patients with ENL and 30 non-reactional LL patient controls at ALERT Hospital, Ethiopia. Blood and skin biopsy samples were obtained from each patient before and after prednisolone treatment. Peripheral blood mononuclear cells from patients with ENL cases and LL controls were cultured with M. leprae whole-cell sonicates (MLWCS), phytohemagglutinin or no stimulation for 6 days. The supernatants were assessed with the enzyme-linked immunosorbent assay for inflammatory and regulatory cytokines. For cytokine gene expression, mRNA was isolated from whole blood and skin lesions and then reverse transcribed into cDNA. The mRNA gene expression was quantified on a Light Cycler using real-time PCR assays specific to TNF, IFN-γ, IL-β, TGF-β, IL-17A, IL-6, IL-8, and IL-10. The ex vivo production of the cytokines: TNF, IFN-γ, IL-1β, and IL-17A was significantly increased in untreated patients with ENL. However, IL-10 production was significantly lower in untreated patients with ENL and significantly increased after treatment. The ex vivo production of IL-6 and IL-8 in patients with ENL did not show statistically significant differences before and after prednisolone treatment. The mRNA expression in blood and skin lesion for TNF, IFN-γ, IL-1β, IL-6, and IL-17A significantly reduced in patients with ENL after treatment, while mRNA expression for IL-10 and TGF-β was significantly increased both in blood and skin lesion after treatment. This is the first study examining the effect of prednisolone on the kinetics of inflammatory and regulatory cytokines in patients with ENL reactions before and after prednisolone treatment. Our findings suggest that prednisolone modulates the pro-inflammatory cytokines studied here either directly or through suppression of the immune cells producing these inflammatory cytokines.

Increased activated memory B-cells in the peripheral blood of patients with erythema nodosum leprosum reactions.

B-cells, in addition to antibody secretion, have emerged increasingly as effector and immunoregulatory cells in several chronic inflammatory diseases. Although Erythema Nodosum Leprosum (ENL) is an inflammatory complication of leprosy, the role of B- cell subsets has never been studied in this patient group. Therefore, it would be interesting to examine the contribution of B-cells in the pathogenesis of ENL. A case-control study design was used to recruit 30 untreated patients with ENL and 30 non-reactional lepromatous leprosy (LL) patient controls at ALERT Hospital, Ethiopia. Peripheral blood samples were obtained before, during and after treatment from each patient. Peripheral blood mononuclear cells (PBMCs) were isolated and used for immunophenotyping of B- cell subsets by flow cytometry. The kinetics of B-cells in patients with ENL before, during and after Prednisolone treatment of ENL was compared with LL patient controls as well as within ENL group. Total B-cells, mature B-cells and resting memory B-cells were not significantly different between patients with ENL reactions and LL controls before treatment. Interestingly, while the percentage of naive B-cells was significantly lower in untreated ENL patients than in LL patient controls, the percentage of activated memory B-cells was significantly higher in these untreated ENL patients than in LL controls. On the other hand, the percentage of tissue-like memory B-cells was considerably low in untreated ENL patients compared to LL controls. It appears that the lower frequency of tissue-like memory B-cells in untreated ENL could promote the B-cell/T-cell interaction in these patients through downregulation of inhibitory molecules unlike in LL patients. Conversely, the increased production of activated memory B-cells in ENL patients could imply the scale up of immune activation through antigen presentation to T-cells. However, the generation and differential function of these memory B-cells need further investigation. The finding of increased percentage of activated memory B-cells in untreated patients with ENL reactions suggests the association of these cells with the ENL pathology. The mechanism by which inflammatory reactions like ENL affecting these memory cells and contributing to the disease pathology is an interesting area to be explored for and could lead to the development of novel and highly efficacious drug for ENL treatment.

T-cell regulation in Erythema Nodosum Leprosum.

Leprosy is a disease caused by Mycobacterium leprae where the clinical spectrum correlates with the patient immune response. Erythema Nodosum Leprosum (ENL) is an immune-mediated inflammatory complication, which causes significant morbidity in affected leprosy patients. The underlying cause of ENL is not conclusively known. However, immune-complexes and cell-mediated immunity have been suggested in the pathogenesis of ENL. The aim of this study was to investigate the regulatory T-cells in patients with ENL. Forty-six untreated patients with ENL and 31 non-reactional lepromatous leprosy (LL) patient controls visiting ALERT Hospital, Ethiopia were enrolled to the study. Blood samples were obtained before, during and after prednisolone treatment of ENL cases. Peripheral blood mononuclear cells (PBMCs) were isolated and used for immunophenotyping of regulatory T-cells by flow cytometry. Five markers: CD3, CD4 or CD8, CD25, CD27 and FoxP3 were used to define CD4+ and CD8+ regulatory T-cells. Clinical and histopathological data were obtained as supplementary information. All patients had been followed for 28 weeks. Patients with ENL reactions had a lower percentage of CD4+ regulatory T-cells (1.7%) than LL patient controls (3.8%) at diagnosis of ENL before treatment. After treatment, the percentage of CD4+regulatory T-cells was not significantly different between the two groups. The percentage of CD8+ regulatory T-cells was not significantly different in ENL and LL controls before and after treatment. Furthermore, patients with ENL had higher percentage of CD4+ T-ells and CD4+/CD8+ T-cells ratio than LL patient controls before treatment. The expression of CD25 on CD4+ and CD8+ T-cells was not significantly different in ENL and LL controls suggesting that CD25 expression is not associated with ENL reactions while FoxP3 expression on CD4+ T-cells was significantly lower in patients with ENL than in LL controls. We also found that prednisolone treatment of patients with ENL reactions suppresses CD4+ T-cell but not CD8+ T-cell frequencies. Hence, ENL is associated with lower levels of T regulatory cells and higher CD4+/CD8+ T-cell ratio. We suggest that this loss of regulation is one of the causes of ENL.

New Insight into the Pathogenesis of Erythema Nodosum Leprosum: The Role of Activated Memory T-Cells.

Memory T-cells, particularly, effector memory T cells are implicated in the pathogenesis of inflammatory diseases and may contribute to tissue injury and disease progression. Although erythema nodosum leprosum (ENL) is an inflammatory complication of leprosy, the role of memory T cell subsets has never been studied in this patient group. The aim of this study was at investigate the kinetics of memory T cell subsets in patients with ENL before and after prednisolone treatment. A case–control study design was used to recruit 35 untreated patients with ENL and 25 non-reactional lepromatous leprosy (LL) patient controls at ALERT Hospital, Ethiopia. Venous blood samples were obtained before, during, and after treatment from each patient. Peripheral blood mononuclear cells (PBMCs) were isolated and used for immunophenotyping of T cell activation and memory T-cell subsets by flow cytometry. The kinetics of these immune cells in patients with ENL before and after treatment were compared with LL patient controls as well as within ENL cases at different time points. The median percentage of CD3+, CD4+, and CD8+ T-cells expressing activated T-cells were significantly higher in the PBMCs from patients with ENL than from LL patient controls before treatment. The median percentage of central and activated memory T-cells was significantly increased in patients with ENL compared to LL patient controls before treatment. Interestingly, patients with ENL had a lower percentage of naïve T cells (27.7%) compared to LL patient controls (59.5%) (P < 0.0001) before treatment. However, after prednisolone treatment, patients with ENL had a higher median percentage of naïve T-cells (43.0%) than LL controls (33.0%) (P < 0.001). The median percentage of activated T-cells (effector memory and effector T-cells) was significantly increased in patients with ENL (59.2%) before treatment compared to after treatment with prednisolone (33.9%) (P < 0.005). This is the first work which has shown T-cell activation and the different subsets of memory T cells in untreated patients with ENL. Consequently, this study delineates the role of T-cell activation in the pathogenesis of ENL reaction and challenges the long-standing dogma of immune complex as a sole etiology of ENL reaction.

A prospective randomised, double-blind, placebo controlled trial on the effect of extended clofazimine on Erythema Nodosum Leprosum (ENL) in multibacillary (MB) leprosy

Objective: The objective of this study was to assess whether additional clofazimine could lower the incidence and/or severity of ENL reactions in leprosy. Methods: Multibacillary subjects, with a BI of over 4+ at any site, were recruited at the Cebu Skin Clinic. On completion of 12 months MB-MDT, which included clofazimine, subjects were randomised to receive either an additional 12 months of clofazimine (100 mg per day) or placebo, and were monitored during that period and for a further period of 12 months afterwards. Results: One hundred subjects were recruited, but only 25 developed ENL during the trial, which was far fewer than expected. On some measures, including deaths likely to be related to steroid intake, the placebo group appeared to have more severe ENL, but none of the differences were statistically significant. Conclusion: No beneficial preventive effect of clofazimine has been demonstrated in this small study. Any further study would require a larger sample size, but the target group could be narrowed to patients who have already had one episode of ENL. Clofazimine remains difficult to source outside its routine availability within MDT.

Experiences with Thalidomide for Erythema Nodosum Leprosum– a retrospective study

Thalidomide is well known as a steroid sparing drug in Erythema Nodosum leprosum (ENL) reaction in leprosy. There is no guideline as to when it should be offered to patients. Documentation of ENL presentation with its morbidity before and after with patient profile can be a baseline to develop a selection criteria as to when thalidomide should be started to reduce steroid related morbidly. Chart and electronic record review was done. 427 ENL patients attended the hospital from 2010 to 2014. 73 patients (67 males six females) were treated with thalidomide. 77% (56) patients were in the age group of 16–45 yrs. 16% (12) were dependent and 39% (29) were taking steroids at presentation. 82% (60) became dependent while on treatment. Ninety five percent were chronic or recurrent ENLs and 73% (53) had moderate to severe ENLs over 49 median months. Steroid induced morbidities were (Cushingoid features 42%, diabetes 21%, infections GI 42%, genitourinary 26%, cataract 23%). There was 11% mortality. Further studies are recommended to diagnose steroid dependence early to prevent serious adverse effects.

Leprosy Reactions Show Increased Th17 Cell Activity and Reduced FOXP3+ Tregs with Concomitant Decrease in TGF-β and Increase in IL-6.

Background50% of leprosy patients suffer from episodes of Type 1/ reversal reactions (RR) and Type 2/ Erythema Nodosum Leprosum (ENL) reactions which lead to morbidity and nerve damage. CD4+ subsets of Th17 cells and CD25+FOXP3+ regulatory T cells (Tregs) have been shown to play a major role in disease associated immunopathology and in stable leprosy as reported by us and others. The aim of our study was to analyze their role in leprosy reactions.Methodology and Principle FindingsQuantitative reverse transcribed PCR (qPCR), flowcytometry and ELISA were used to respectively investigate gene expression, cell phenotypes and supernatant levels of cytokines in antigen stimulated PBMC cultures in patients with stable disease and those undergoing leprosy reactions. Both types of reactions are associated with significant increase of Th17 cells and associated cytokines IL-17A, IL-17F, IL-21, IL-23 and chemokines CCL20, CCL22 as compared to matching stable forms of leprosy. Concurrently patients in reactions show reduction in FOXP3+ Treg cells as well as reduction in TGF-β and increase in IL-6. Moreover, expression of many T cell markers, cytokines, chemokines and signaling factors were observed to be increased in RR as compared to ENL reaction patients.ConclusionsPatients with leprosy reactions show an imbalance in Th17 and Treg populations. The reduction in Treg suppressor activity is associated withhigherTh17cell activity. The combined effect of reduced TGF-β and enhanced IL-6, IL-21 cytokines influence the balance between Th17 or Treg cells in leprosy reactions as reported in the murine models and autoimmune diseases. The increase in Th17 cell associated cytokines may contribute to lesional inflammation.