Visceral leishmaniasis (VL) is characterized by fever, splenomegaly, hepatomegaly and anemia. Leishmania amastigotes can infect mononuclear phagocytes of spleen, liver, bone marrow and lymph nodes (LN). Similarly, spleen and LN are critical reservoirs for maintenance of HIV, contributing to the virus persistence. Patients with VL co-infected with HIV/AIDS often remain with relapses, multiple hospital readmissions, and low quality of life despite drug therapy. We have previously reported the benefits of splenectomy as rescue therapy in patients with chronic drug-refractory VL. Herein, we described the parasitism and anatomopathological features of spleen, LN and skin in patients with VL-HIV/AIDS who underwent laparoscopic splenectomy. This study aims contribute to the understanding of host-parasite interactions, disease progression, reactivation and VL- transmission. A total of 10 patients donated their spleen, and three patients also donated samples from mesenteric LN and skin from the incision site. They were confirmed to VL and HIV infection, not responsive in two cycles of Amphotericin B with six months interval, submitted to secondary prophylaxis (Amphotericin B biweekly) and adherent to highly active anti- retroviral therapy. After chirurgical procedure, spleens were weighted and 0,3-cm3 splenic tissues sections, 0,5-cm3 skin sections, complete mesenteric LN were collected and fixed in an alcoholic acid formalin solution. Tissues were paraffin-embedded, and sections were smeared with hematoxylin- eosin prior to microscopy examination. Structural changes of the spleen were systemically classified in three categories. Spleen type 1: well- organized white pulp with discernible peri-arteriolar lymphocyte sheath, germinal center, mantle zone and marginal zone. Spleen type 2: slightly disorganized white pulp with either hyperplastic or hypoplastic changes leading to a loss in definition of the limits between white pulp regions. Spleen type 3: moderately to extensively disorganized white pulp with poorly discernible regions. Follicular structure hardly distinct from the red pulp and T-cell areas. Lymph nodes and skin were evaluated for the presence of parasites, hemosiderin deposits (siderosis) and cellularity. Spleen weigh ranged from 295 g to 1882 g, with mean of 794.3 g and median 650 g. Three patients (30%) had spleen type 1, five (50%) spleen type 2 and two (20%) spleen type 3. All spleens had massive presence of Leishmania amastigotes. Three mesenteric LN had intense presence of parasites and one perisplenic LN was negative for Leishmania. Remarkable, two out of three skin samples had perivascular macrophages containing intense amastigote parasitism. Additionally, one patient had post kala-azar dermal leishmaniasis after splenectomy, with confirmed skin parasitism. Siderosis was present in all LN and four spleens. Disorganization of the spleen compartments is associated with more severe VL-presentation. Combine with the increasing of spleen size, the loss of lymphoid tissue architecture leads to a defective humoral and cellular response. Heavily parasitized mesenteric LN indicates its importance to the parasite persistence after treatment. Cutaneous parasitism infections may act as reservoirs contributing to disease reactivation and allowing anthroponotic transmission. Abnormal tissue retention of iron may underlie sideropenic anemia. We showed that the disorganization of splenic microenvironments is relevant to human-VL, thus splenectomy may improve the clinical course of patients with chronic drug-refractory VL.