Comparison of physiological, cytopathogenic and immunological properties between two environmental isolates of Acanthamoeba spp.

The aim of this study was to determine whether pathogenic and less-pathogenic isolates of environmental Acanthamoeba exhibit differences in adhesion to human erythrocytes. Based on physiological properties of temperature, tolerance, and rapid growth, Acanthamoeba were divided into pathogenic and less-pathogenic isolates. Acanthamoeba were tested for their ability to produce cytopathic effects (CPE) using two human cell lines, HEp-2 and KB cells. Both ameba isolates caused CPE to both cell lines with the same pattern without significant difference. Human erythrocytes from 20 healthy volunteers were used to study the erythrocyte reactivity of Acanthamoeba by co-incubation with trophozoites. The pathogenic Acanthamoeba exhibited significantly higher erythrocyte adhesion as compared to the less-pathogens (p<0.05). Erythrocyte activity occurred in the presence of plasma in all blood samples, suggesting the role of plasmatic components and contact-dependent mechanisms to produce host cell cytotoxicity. The present results showed correlation between the physiological properties and erythrocyte reactivity of Acanthamoeba.

Characterization of inhibitory anti-Duffy binding protein II immunity: approach to Plasmodium vivax vaccine development in Thailand.

Plasmodium vivax Duffy binding protein region II (DBPII) is an important vaccine candidate for antibody-mediated immunity against vivax malaria. A significant challenge for vaccine development of DBPII is its highly polymorphic nature that alters sensitivity to neutralizing antibody responses. Here, we aim to characterize naturally-acquired neutralizing antibodies against DBPII in individual Thai residents to give insight into P. vivax vaccine development in Thailand. Anti-DBPII IgG significantly increased in acute vivax infections compared to uninfected residents and naive controls. Antibody titers and functional anti-DBPII inhibition varied widely and there was no association between titer and inhibition activity. Most high titer plasmas had only a moderate to no functional inhibitory effect on DBP binding to erythrocytes, indicating the protective immunity against DBPII binding is strain specific. Only 5 of 54 samples were highly inhibitory against DBP erythrocyte-binding function. Previously identified target epitopes of inhibitory anti-DBPPII IgG (H1, H2 and H3) were localized to the dimer interface that forms the DARC binding pocket. Amino acid polymorphisms (monomorphic or dimorphic) in H1 and H3 protective epitopes change sensitivity of immune inhibition by alteration of neutralizing antibody recognition. The present study indicates Thai variant H1.T1 (R308S), H3.T1 (D384G) and H3.T3 (K386N) are the most important variants for a DBPII candidate vaccine needed to protect P. vivax in Thai residents.