title: Serum-mediated activation of macrophages reflects TcVac2 vaccine efficacy against Chagas disease.
abstract: Chagas disease is endemic in Latin America and an emerging infectious disease in the United States. No effective treatments are available. The TcG1, TcG2, and TcG4 antigens are highly conserved in clinically relevant Trypanosoma cruzi isolates and are recognized by B and T cells in infected hosts. Delivery of these antigens as a DNA prime/protein boost vaccine (TcVac2) elicited lytic antibodies and type 1 CD8(+) T cells that expanded upon challenge infection and provided >90% control of parasite burden and myocarditis in chagasic mice. Here we determined if peripheral blood can be utilized to capture the TcVac2-induced protection from Chagas disease. We evaluated the serum levels of T. cruzi kinetoplast DNA (TckDNA), T. cruzi 18S ribosomal DNA (Tc18SrDNA), and murine mitochondrial DNA (mtDNA) as indicators of parasite persistence and tissue damage and monitored the effect of sera on macrophage phenotype. Circulating TckDNA/Tc18SrDNA and mtDNA were decreased by >3- to 5-fold and 2-fold, respectively, in vaccinated infected mice compared to nonvaccinated infected mice. Macrophages incubated with sera from vaccinated infected mice exhibited M2 surface markers (CD16, CD32, CD200, and CD206), moderate proliferation, a low oxidative/nitrosative burst, and a regulatory/anti-inflammatory cytokine response (interleukin-4 [IL-4] plus IL-10 > tumor necrosis factor alpha [TNF-α]). In comparison, macrophages incubated with sera from nonvaccinated infected mice exhibited M1 surface markers, vigorous proliferation, a substantial oxidative/nitrosative burst, and a proinflammatory cytokine response (TNF-α ≫ IL-4 plus IL-10). Cardiac infiltration of macrophages and TNF-α and oxidant levels were significantly reduced in TcVac2-immunized chagasic mice. We conclude that circulating TcDNA and mtDNA levels and macrophage phenotype mediated by serum constituents reflect in vivo levels of parasite persistence, tissue damage, and inflammatory/anti-inflammatory state and have potential utility in evaluating disease severity and efficacy of vaccines and drug therapies.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Shivali
authors.initials: S
authors.lastname: Gupta
authors.forename: Trevor S
authors.initials: TS
authors.lastname: Silva
authors.forename: Jessica E
authors.initials: JE
authors.lastname: Osizugbo
authors.forename: Laura
authors.initials: L
authors.lastname: Tucker
authors.forename: Heidi M
authors.initials: HM
authors.lastname: Spratt
authors.forename: Nisha J
authors.initials: NJ
authors.lastname: Garg
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000276:immunology; D014355:Chagas Disease / Q000276:immunology / Q000517:prevention & control*; D016207:Cytokines / Q000378:metabolism; D018105:DNA, Kinetoplast / Q000097:blood; D004272:DNA, Mitochondrial / Q000097:blood; D005260:Female; D008262:Macrophage Activation / Q000276:immunology*; D020782:Matrix Metalloproteinases / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016052:Protozoan Vaccines / Q000276:immunology*; D012337:RNA, Ribosomal, 18S / Q000097:blood; D014349:Trypanosoma cruzi / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D015703:Antigens, CD; D016207:Cytokines; D018105:DNA, Kinetoplast; D004272:DNA, Mitochondrial; D016052:Protozoan Vaccines; D012337:RNA, Ribosomal, 18S; D019444:Vaccines, DNA; D020782:Matrix Metalloproteinases