In Vitro Differentiation of Human Dendritic Cells and their Markers in Leishmania Infection.

Leishmaniasis comprises a collection of clinical manifestations associated with the infection of obligate intracellular protozoans, Leishmania. The life cycle of Leishmania parasites consists of two alternating life stages (amastigotes and promastigotes), during which parasites reside within either arthropod vectors or vertebrate hosts, respectively. Notably, the complex interactions between Leishmania parasites and several cells of the immune system largely influence the outcome of infection. Importantly, although macrophages are known to be the main host niche for Leishmania replication, parasites are also phagocytosed by other innate immune cells, such as neutrophils and dendritic cells (DCs). DCs play a major role in bridging the innate and adaptive branches of immunity and thus orchestrate immune responses against a wide range of pathogens. The mechanisms by which Leishmania and DCs interact remain unclear and involve aspects of pathogen capture, the dynamics of DC maturation and activation, DC migration to draining lymph node (dLNs), and antigen presentation to T cells. Although a large body of studies support the notion that DCs play a dual role in modulating immune responses against Leishmania, the participation of these cells in susceptibility or resistance to Leishmania remains poorly understood. After infection, DCs undergo a maturation process associated with the upregulation of surface major histocompatibility complex (MHC) II, in addition to costimulatory molecules (namely, CD40, CD80, and CD86). Understanding the role of DCs in infection outcome is crucial to developing therapeutic and prophylactic strategies to modulate the immune response against Leishmania. This paper describes a method for the characterization of Leishmania-DC interaction. This detailed protocol provides guidance throughout the steps of DC differentiation, the characterization of cell surface molecules, and infection protocols, allowing scientists to investigate DC response to Leishmania infection and gain insight into the roles played by these cells in the course of infection.

Cell Migration and Cell Adhesion Assays to Investigate Leishmania-Host Cell Interaction.

Leishmania is an intracellular protozoan parasite that causes a broad spectrum of clinical manifestations, ranging from self-resolving localized cutaneous lesions to a highly fatal visceral form of the disease. An estimated 12 million people worldwide are currently infected, and another 350 million face risk of infection. It is known that host cells infected by Leishmania parasites, such as macrophages or dendritic cells, can migrate to different host tissues, yet how migration contributes to parasite dissemination and homing remains poorly understood. Therefore, assessing these parasites' ability to modulate host cell response, adhesion, and migration will shed light on mechanisms involved in disease dissemination and visceralization. Cellular migration is a complex process in which cells undergo polarization and protrusion, allowing them to migrate. This process, regulated by actin and tubulin-based microtubule dynamics, involves different factors, including the modulation of cellular adhesion to the substrate. Cellular adhesion and migration processes have been investigated using several models. Here, we describe a method to characterize the migratory aspects of host cells during Leishmania infection. This detailed protocol presents the differentiation and infection of dendritic cells, the analysis of host cell motility and migration, and the formation of adhesion complexes and actin dynamics. This in vitro protocol aims to further elucidate mechanisms involved in Leishmania dissemination within vertebrate host tissues and can also be modified and applied to other cell migration studies.

Leishmania-Induced Dendritic Cell Migration and Its Potential Contribution to Parasite Dissemination.

Leishmania, an intracellular parasite species, causes lesions on the skin and in the mucosa and internal organs. The dissemination of infected host cells containing Leishmania is crucial to parasite survival and the establishment of infection. Migratory phenomena and the mechanisms underlying the dissemination of Leishmania-infected human dendritic cells (hDCs) remain poorly understood. The present study aimed to investigate differences among factors involved in hDC migration by comparing infection with visceral leishmaniasis (VL) induced by Leishmaniainfantum with diverse clinical forms of tegumentary leishmaniasis (TL) induced by Leishmaniabraziliensis or Leishmania amazonensis. Following the infection of hDCs by isolates obtained from patients with different clinical forms of Leishmania, the formation of adhesion complexes, actin polymerization, and CCR7 expression were evaluated. We observed increased hDC migration following infection with isolates of L. infantum (VL), as well as disseminated (DL) and diffuse (DCL) forms of cutaneous leishmaniasis (CL) caused by L. braziliensis and L. amazonensis, respectively. Increased expression of proteins involved in adhesion complex formation and actin polymerization, as well as higher CCR7 expression, were seen in hDCs infected with L. infantum, DL and DCL isolates. Together, our results suggest that hDCs play an important role in the dissemination of Leishmania parasites in the vertebrate host.

Prevalência de mamoplastia redutora feminina no Brasil de 2015 a 2019 / Prevalence of female reduction mammoplasty in Brazil from 2015 to 2019

RESUMO Introdução: A mamoplastia redutora feminina visa restaurar o volume fisiológico das mamas em casos de hipertrofia mamária, de modo a manter a simetria e com um número mínimo de complicações. Embora tal procedimento seja usual, a literatura carece de dados epidemiológicos e descritivos acerca da cirurgia de redução de mama. Desse modo, esse estudo tem como objetivo descrever a prevalência da mamoplastia redutora no Brasil dos anos de 2015 a 2019; comparar a prevalência de mamoplastia redutora entre as regiões do Brasil de 2015-2018 com a do ano de 2019; e avaliar a relação entre a renda familiar regional e a prevalência da mamoplastia redutora feminina no país de 2015-2019. Métodos: Estudo ecológico de base populacional cujos dados foram coletados por meio do Departamento de Informática do Sistema Único de Saúde e censos demográficos do Instituto Brasileiro de Geografia e Estatística. Foram coletados dados de 2015 a 2019. Resultados: A prevalência de mamoplastia redutora no Brasil no ano de 2019 foi de 0.08%, enquanto que nos anos de 2015 a 2018, essa prevalência foi de 0.07%. Não houve diferença estatisticamente significante quando comparadas as prevalências regionais dos anos de 2015 a 2018 com a prevalência de 2019. As prevalências de mamoplastia redutora nas regiões do Brasil de 2015-2019 apresentaram correlação positiva moderada com renda domiciliar média per capita. Conclusão: A prevalência de mamoplastia redutora feminina no Brasil apresenta uma tendência de estabilidade temporal ao longo dos anos de 2015-2019 e-m âmbito nacional e regional. Sugere-se uma correlação positiva entre a renda familiar per capita regional e a prevalência desse procedimento.

ABSTRACT Introduction: Female reducing mammoplasty aims to restore the physiological volume of the breasts in cases of breast hypertrophy, to maintain symmetry and with a minimum number of complications. Although such a procedure is usual, the literature lacks epidemiological and descriptive data on breast reduction surgery. Thus, this study aims to describe the prevalence of reducing mammoplasty in Brazil from 2015 to 2019; to compare the prevalence of reducing mammoplasty among the regions of Brazil from 2015-2018 with that of 2019, and to evaluate the relationship between regional family income and the prevalence of female reduction mammoplasty in the country of 2015-2019. Methods: Population-based ecological study whose data were collected through Informatics Department of the Unified Health System and Brazilian Institute of Geography and Statistics demographic censuses. Data were collected from 2015 to 2019. Results: The prevalence of reducing mammoplasty in Brazil in 2019 was 0.08%, while from 2015 to 2018, this prevalence was 0.07%. There was no statistically significant difference compared to the regional prevalence from 2015 to 2018 with the prevalence of 2019. The prevalence of reducing mammoplasty in the regions of Brazil from 2015-2019 showed a moderate positive correlation with average household income per capita. Conclusion: The prevalence of female reduction mammoplasty in Brazil shows a trend of temporal stability over the years 2015-2019 at national and regional levels. A positive correlation between regional per capita family income and the prevalence of this procedure is suggested.