Biodegradable microspheres containing leukotriene B(4) and cell-free antigens from Histoplasma capsulatum activate murine bone marrow-derived macrophages.
Because of the potential protective role of leukotrienes (LTs) in histoplasmosis and the therapeutic and prophylactic effects of cell-free antigens from Histoplasmacapsulatum (CFAgs), the aim of this study was to develop and characterise biodegradable LTB(4)/CFAgs-loaded microspheres (MS) that could promote cellular activation for future immunisation purposes. LTB(4)/CFAgs-loaded MS that were developed through a double emulsion/extraction process were characterised according to their size, zeta potential, morphology, entrapment efficiency and in vitro release kinetics. We evaluated the uptake of LTB(4)/CFAgs-loaded MS by bone marrow derived-macrophages (BMDM). The TNF-α and chemokines, and nitrite production, in the supernatant of BMDM cultures were analysed by enzyme-linked immunosorbent assay (ELISA) and Griess reaction, respectively. We found an instantaneous release of CFAgs and a prolonged release of LTB(4) from the poly-(d,l-lactide-co-glycolide) (PLGA) MS. The microencapsulation process did not alter the zeta potential nor the spherical morphology of the MS. The appropriate size of the LTB(4)/CFAgs-loaded MS (smaller than 10μm) enabled the efficient uptake by BMDM and also induced TNF-α, CXCL1/KC, CCL2/MCP-1, CCL5/RANTES and nitrite oxide release by these cells. In conclusion, the biodegradable LTB(4)/CFAgs-loaded MS were able to efficiently activate murine BMDM and thereby have the potential to be used in an effective vaccine against H. capsulatum infection.
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Universidade de São Paulo, Ribeirão Preto, Brazil.
Daiane F
DF
Dos Santos
Claudia S
CS
Bitencourt
Guilherme M
GM
Gelfuso
Priscilla A T
PA
Pereira
Patrícia R M
PR
de Souza
Carlos A
CA
Sorgi
Roberto
R
Nicolete
Lúcia H
LH
Faccioli
D000818:Animals; D000946:Antigens, Fungal / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D018925:Chemokines / Q000276:immunology; D006658:Histoplasma / Q000276:immunology* / Q000378:metabolism; D019344:Lactic Acid; D007975:Leukotriene B4 / Q000276:immunology* / Q000378:metabolism; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008863:Microspheres; D009569:Nitric Oxide / Q000276:immunology; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D000946:Antigens, Fungal; D018925:Chemokines; D014409:Tumor Necrosis Factor-alpha; D007975:Leukotriene B4; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D009569:Nitric Oxide; D019344:Lactic Acid