Electrolyzed Oxidizing Water Modulates the Immune Response in BALB/c Mice Experimentally Infected with Trypanosoma cruzi

Abstract

Chagas disease is a major public health problem in Latin America. The mixed Th1/Th2 immune response is required against Trypanosoma cruzi. Electrolyzed oxidizing water (EOW) has been shown to have germicidal efficacy. The objective of this study was to evaluate the EOW effectiveness in T. cruzi-infected BALB/c mice clinically, immunologically, and histologically. The severity of the infection was assessed by parasitaemia, general health condition, mortality, mega syndromes, and histological lesions. IgG, TNF-alpha, IFN-gamma, and IL-1 beta levels were quantified. The EOW administration showed a beneficial effect on parasitaemia, general physical condition, and mortality. High levels of IgG1 at 50 days postinfection were observed. Prophylactic EOW treatment was able to induce a predominantly TH1 immune response based on an IgG2a levels increase at the late acute phase, and a 10-fold increase of IFN-gamma in whole acute phase. EOW was able to control the acute phase infection as effectively as benznidazole. Splenomegaly was caused by EOW treatment and lymphadenopathy was stimulated by T. cruzi infection in all groups. Severe tissue damage was not prevented by EOW treatments. Moderate efficacy may be due to immunomodulatory properties and not to a direct toxic effect on the parasite.

Mycobacterium bovis BCG as immunostimulating agent prevents the severe form of chronic experimental Chagas disease

“…On the other hand, it was established that vaccination with BCG plus dead Leishmania promastigotes reduced the acute infection by T. cruzi in a murine model, increasing survival time and decreasing parasitemia and mortality ( 33 ). The results of parasitemia in this study are comparable with previous ones ( 30 , 34 ), in which two peaks of parasitemia were described with magnitudes of 10 6 parasites/mL of blood in a similar infection with the Ninoa strain intraperitoneally with 150 BT. Other research groups ( 35 , 36 ) have reported peaks of parasitemia that reach levels of 10 6 parasites/mL of blood in BALB/c mice infected with two Mexican strains Querétaro and Ninoa belonging to DTU TcI, in which the infection inoculum was around 10 4 BT.…”

“…Evaluation of humoral immune response by the search of anti- T. cruzi IgG antibodies in the sera of BCG-immunized/ T. cruzi -infected and T. cruzi -infected mice, as well as IgG1 and IgG2a subclasses for knowing if a Th1, Th2, or a mixed Th1/Th2-type immune response is triggered, was performed by ELISA as described previously ( 30 ): 1 μg/mL of a whole protein extract of T. cruzi INC-9 isolate in 200 μL of carbonate buffer (NaCO 3 /NaHCO 3 , pH 9.6) was added to each well of Costar 96-well plates (Corning, USA, catalog 2592) and incubated at 37°C for 1 h. The wells were washed five times with 300 μL of 0.05% PBS-T, and 200 μL of BSA was placed for incubation at 37°C for 20 min. Serum samples were diluted in this blocking solution by adding 1 μL of mouse sera (1:200 in PBS) and incubated at 37°C for 1 h. The plates were washed five times with 300 μL of PBS-T. After washing, the peroxidase-labeled rabbit anti-mouse IgG and IgG subclasses (IgG1 and IgG2a) of secondary antibodies (Novus Biologicals, Centennial, CO, USA) were added at 1:10,000 dilution in PBS-T and incubated for 1 h at room temperature.…”

Nitazoxanide: A Drug Repositioning Compound with Potential Use in Chagas Disease in a Murine Model

“…Determination of immunoglobulin G and its subclasses was performed by enzyme-linked immunosorbent assay (ELISA) using the sera obtained for each group, as previously described [ 16 ]. Anti-mouse peroxidase-conjugated antibodies (Novus Biologicals, Littleton, CO, USA) and anti-mouse IgG, IgG1, and IgG2a were used.…”

“…Commercial ELISA kits (Enzo Life Sciences, Inc. ® , Farmingdale, NY, USA and R&D Systems, Inc. ® , Minneapolis, MN, USA) were used to determine levels of some proinflammatory cytokines (TNF-α, IFN-γ, and IL-1β,) in the mouse sera at 40 and 60 days after infection, according to the manufacturer’s instructions and as described previously [ 16 ].…”

“…The mice were euthanized on day 60 post-infection and the following organs were collected: the heart, spleen, esophagus, large intestine, small intestine, brain, skeletal muscle, and peripheral lymph nodes (poplitei). After histological processing, the tissue sections were stained with hematoxylin and eosin for analysis, as previously described [ 16 ].…”