Immunogens in Balamuthia mandrillaris: a proteomic exploration.

Balamuthia mandrillaris is the causative agent of granulomatous amoebic encephalitis, a rare and often fatal infection affecting the central nervous system. The amoeba is isolated from diverse environmental sources and can cause severe infections in both immunocompromised and immunocompetent individuals. Given the limited understanding of B. mandrillaris, our research aimed to explore its protein profile, identifying potential immunogens crucial for early granulomatous amoebic encephalitis diagnosis. Cultures of B. mandrillaris and other amoebas were grown under axenic conditions, and total amoebic extracts were obtained. Proteomic analyses, including two-dimensional electrophoresis and mass spectrometry, were performed. A 50-kDa band showed a robust recognition of antibodies from immunized BALB/c mice; peptides contained in this band were matched with elongation factor-1 alpha, which emerged as a putative key immunogen. Besides, lectin blotting revealed the presence of glycoproteins in B. mandrillaris, and confocal microscopy demonstrated the focal distribution of the 50-kDa band throughout trophozoites. Cumulatively, these observations suggest the participation of the 50-kDa band in adhesion and recognition mechanisms. Thus, these collective findings demonstrate some protein characteristics of B. mandrillaris, opening avenues for understanding its pathogenicity and developing diagnostic and therapeutic strategies.

In Vitro Evaluation of Combined Commercialized Ophthalmic Solutions Against Acanthamoeba Strains.

Acanthamoeba is a free-living amoebae genus which is present worldwide in natural and artificial environments. These amoebae are clinically important as causative agents of diseases in humans and other animals such as a fatal encephalitis or a sight threatening Acanthamoeba keratitis (AK). Lately; studies have focused on the search of novel therapeutic options for AK but also to prevent infections. Furthermore; the evaluation of commercialized products seems to be an option for this case since not clinical assays would be required. Thus; we aimed to test the amoebicidal activity of different mixtures of two commercial ophthalmic solutions: Systane® Ultra; which has already shown anti-Acanthamoeba properties; and Naviblef® Daily Care. In addition, we tested their cytotoxic effect against murine macrophages. At the individual level; Naviblef® Daily Care showed to be the most active product against Acanthamoeba spp. Nevertheless; the combinations of Systane® Ultra and Naviblef® Daily Care; showed an improvement in the activity against trophozoites and cysts of Acanthamoeba castellanii Neff. Moreover; the concentration necessary to generate cytotoxic effect against murine macrophages (J774.1) was much higher than the required for the amoebicidal and cysticidal effect achieved in the most effective mixtures.

Detection of serum antibodies in children and adolescents against Balamuthia mandrillaris, Naegleria fowleri and Acanthamoeba T4.

The presence of free-living amoebae of the genera Naegleria, Acanthamoeba and Balamuthia, which contain pathogenic species for humans and animals, has been demonstrated several times and in different natural aquatic environments in the northwest of Mexico. With the aim of continuing the addition of knowledge about immunology of pathogenic free-living amoebae, 118 sera from children and adolescents, living in three villages, were studied. Humoral IgG response against B. mandrillaris, N. fowleri and Acanthamoeba sp. genotype T4, was analyzed in duplicate to titers 1: 100 and 1: 500 by enzyme-linked immunosorbent assay (ELISA). Children and adolescents ages ranged between 5 and 16 years old, with a mean of 9 years old, 55% males. All tested sera were positive for the 1: 100 dilution, and in the results obtained with the 1: 500 dilution, 116 of 118 (98.3%) were seropositive for N. fowleri, 101 of 118 (85.6%) were seropositive for Acanthamoeba sp. genotype T4, and 43 of 118 (36.4%) were seropositive for B. mandrillaris. The statistical analysis showed different distributions among the three communities and for the three species of pathogenic free-living amoebae, including age. Lysed and complete cells used as Balamuthia antigens gave differences in seropositivity.