Flow cytometry and fluorimetry in diseases caused by protozoa

• Grant number: 13/04676-9
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2013
• End date: July 31, 2016
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Heitor Franco de Andrade Junior
• Grantee: Heitor Franco de Andrade Junior
• Host Institution: Instituto de Medicina Tropical de São Paulo (IMT). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Andrés Jimenez Galisteo Jr ; Luciana Regina Meireles Jaguaribe Ekman ; Nanci Do Nascimento

Abstract

We studied the diagnosis and pathogenic process in endemic human and experimental infections caused by protozoa and other infectious agents, especially toxoplasmosis, mostly devoted to the detection of infections by specific antibodies and the evaluation of an oral vaccine with irradiated parasites for felids. In the detection of specific antibodies, the main tool is the ELISA and their variations, which are difficult technique to quantify antibodies due its bi-exponential character. Studies of vaccine effect by cell proliferation to antigen challenge are limited if you use radiotracers, but recently new approaches using highly efficient flow cytometry and fluorometry and we achieved recently core facilities for those techniques. Fluorometric approaches allow better quantification of specific antibodies, with high throughput assays and the use of 11 color flow cytometry will provide a better approach to understand cell proliferation to antigen, with more adequate vaccine control. We intend to develop single well detection of specific antibody class (IgG, IgM e IgA) in human toxoplasmosis in our collection of sera with clinical and serological pre-defined patterns, allowing also better quantification in IgG avidity assays, with possible liquid arrays for use in antenatal diagnosis of T.gondii infection in pregnant women and other risky groups. In the same way in vaccine development, we intend to identify groups of antigen challenged cells by high efficient multiple color flow citometry, looking for identification of markers of efficiency of oral vaccine in experimental mouse models. Those projects are the best way to change a technology basis of our laboratory, essential for the new studies devoted to solve the important scientific challenges in human endemic diseases caused by protozoa, allowing both quantitative and qualitative progression in this important field, using preventive, vaccine screening, experimental or epidemiological approaches. (AU)

Scientific publications

(6)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DA COSTA, ANDREA; DE CARVALHO, CAMILA APARECIDA; DO NASCIMENTO, NANCI; DE ANDRADE JR, HEITOR FRANCO. Scavenger receptors mediate increased uptake of irradiated T.gondii extracts by J774 macrophages. International Journal of Radiation Biology, v. 99, n. 10, p. 11-pg., 2023-03-10. (13/04676-9)

FIALHO SAMPAIO, BARBARA CARVALHO; RODRIGUES, JAQUELINE POLIZELI; MEIRELES, LUCIANA REGINA; DE ANDRADE JUNIOR, HEITOR FRANCO. Measles, rubella, mumps and Toxoplasma gondii antibodies in saliva of vaccinated students of schools and universities in Sao Paulo City, Brazil. Brazilian Journal of Infectious Diseases, v. 24, n. 1, p. 51-57, JAN-FEB 2020. (13/04676-9)

ZORGI, NAHIARA ESTEVES; GALISTEO, JR., ANDRES JIMENEZ; SATO, MARIA NOTOMI; DO NASCIMENTO, NANCI; DE ANDRADE, JR., HEITOR FRANCO. Immunity in the spleen and blood of mice immunized with irradiated Toxoplasma gondii tachyzoites. MEDICAL MICROBIOLOGY AND IMMUNOLOGY, v. 205, n. 4, p. 297-314, AUG 2016. (13/04676-9)

RODRIGUES, J. P.; ANDRADE, JR., H. F.. Efficient duplex solid-phase fluorescent assay (dFISA) for the simultaneous detection of specific anti-T. gondii IgG and IgM due to refined conjugates. JOURNAL OF IMMUNOLOGICAL METHODS, v. 420, p. 11-17, MAY 2015. (13/04676-9)

RODRIGUES, JAQUELINE POLIZELI; DE ANDRADE JUNIOR, HEITOR FRANCO. fficiency of a Single well IgG, IgM and IgA Anti T. gondii Fluorimetric Assay for Pre-natal Screening for Congenital Toxoplasmosi. Journal of Fluorescence, v. 32, n. 2, p. 661-667, MAR 2022. (13/04676-9)

DA COSTA, ANDREA; DE ANDRADE JR, HEITOR FRANCO. Toxoplasma gondii in CD36-/- mice shows lethal infection and poor immunization with probable macrophage immune defects. Parasitology Research, v. N/A, p. 9-pg., 2023-03-29. (13/04676-9, 14/17029-4)