Kinetic analysis of ex vivo human blood infection by Leishmania.

PLoS Neglected Tropical Diseases
Inmaculada MorenoAlfredo Toraño

Abstract

The leishmanioses, vector-borne diseases caused by the trypanosomatid protozoan Leishmania, are transmitted to susceptible mammals by infected phlebotomine sand flies that inoculate promastigotes into hemorrhagic pools created in host skin. We assumed that promastigotes are delivered to a blood pool, and analyzed early promastigote interactions (0-5 min) with host components, which lead to parasite endocytosis by blood leukocytes, and to host infection. Promastigotes were incubated with NHS or with heparinized blood in near-physiological conditions, and we used cell radioimmunoassay and flow cytometry to measure the on-rate constants (k(+1)) of promastigote interactions with natural opsonins and erythrocytes. We obtained quantitative data for parasitized cells to determine the time-course of promastigote binding and internalization by blood leukocytes. In these reactions, promastigotes bind natural opsonins, immune adhere to erythrocytes and activate complement cytolysis, which kills approximately 95% of promastigotes by 2 min post-infection. C3-promastigote binding is a key step in opsonization; nascent C3-promastigotes are the substrate for two simultaneous reactions, C3-promastigote immune adherence (IA) to erythrocytes and ...Continue Reading

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Citations

Nov 26, 2010·Dermatologic Clinics·Ron Birnbaum, Noah Craft
Nov 5, 2014·Frontiers in Immunology·Pedro CecílioAnabela Cordeiro da Silva
May 6, 2016·Parasites & Vectors·Juliana Perrone de MenezesBruno da Rocha-Azevedo
May 8, 2016·Journal of Leukocyte Biology·Thiago DeSouza-VieiraElvira M Saraiva
Oct 8, 2013·PLoS Neglected Tropical Diseases·Julien LemairePatricia Renard
Jun 13, 2020·Frontiers in Cellular and Infection Microbiology·Priyank JaiswalSumi Mukhopadhyay

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Methods Mentioned

BETA
FCS
Assay
flow cytometry
by flow
light microscopy

Software Mentioned

CELLQuest
SigmaPlot

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