IgG3 and IL10 are effective biomarkers for monitoring therapeutic effectiveness in Post Kala-Azar Dermal Leishmaniasis

Abstract

Background The assessment of chemotherapeutic responses in Post Kala-azar Dermal Leishmaniasis (PKDL), especially its macular form is challenging, emphasizing the necessity for ‘test of cure’ tools. This study explored the diagnostic and prognostic potential of IgG subclasses and associated cytokines for monitoring the effectiveness of chemotherapy in PKDL. Methods Participants included PKDL cases at (a) disease presentation, (b) immediately at the end of treatment (12 weeks for Miltefosine or 3 weeks for Liposomal Amphotericin B, LAmB and (c) at any time point 6 months later, for estimating anti-leishmanial immunoglobulin (Ig, IgG, IgM, IgG1, IgG2 and IgG3) and cytokines (IL-10, IL-6). Results In PKDL, Ig levels were elevated, with IgG3 and IL-10 being the major contributors. Miltefosine decreased both markers substantially and this decrease was sustained for at least six months. In contrast, LAmB failed to decrease IgG3 and IL-10, as even after six months, their levels remained unchanged or even increased. Conclusions In PKDL, IgG3 and IL-10 proved to be effective predictors of responsiveness to chemotherapy and may be considered as a non invasive alternative for longitudinal monitoring. Author summary Post Kala-azar Dermal Leishmaniasis (PKDL) is a dermal condition that occurs in East Africa and South Asia, the latter in 5–10% of patients after apparent cure from Visceral Leishmaniasis (VL). Till date, conventional knowledge in South Asia was that the polymorphic form of PKDL comprising of macules, papules and nodules was the predominant disease form, constituting 85–90%. However, since 2014, implementation of active-case surveillance led to unearthing of a large number of macular, hypopigmented cases, and was reported to contribute to nearly 50% of the disease burden. In particular, the macular form poses a diagnostic dilemma as microscopically parasites are difficult to identify in their lesions, and repigmentation occurs months after parasite clearance, emphasizing the need for developing non-invasive approaches for measurement of parasite burden. Till date, no formal clinical trial for treatment of PKDL has been undertaken where the parasite load was quantified and treatment remains empirical. This is primarily due to PKDL cases being unwilling to provide a repeat skin biopsy once their lesions have declined. Therefore, in cases where treatment failure occurs, it cannot be precisely identified, and could potentially lead to these cases becoming mobile disease reservoirs, thereby adversely impacting on the ongoing VL elimination programme. This study addressed this critical lacuna, where it was established that in both clinical types of PKDL, circulating levels of IgG3 and IL-10 can be considered as effective markers for monitoring treatment outcome. At disease presentation, the raised levels of IgG subclasses and associated cytokines (IL-10 and IL-6) declined following therapy with Miltefosine, the maximum decrease being with IgG3 along with IL-10; importantly, this decrease was sustained for at least six months. In contrast, LAmB failed to decrease the levels of immunoglobulins and associated cytokines even six months after completion of treatment; in fact the antibody levels either increased or remained unchanged. Taken together, this study has established the potential of IgG3 and IL10 as a non-invasive alternative for monitoring of chemotherapeutic responses in PKDL.