Exploring Onchocerca volvulus Cysteine Protease Inhibitor for Multi-epitope Subunit Vaccine Against Onchocerciasis: An Immunoinformatics Approach

Onchocerciasis, caused by Onchocerca volvulus, affects more than 37 million people worldwide. Despite the progress achieved with mass drug distribution, suitable vaccines against onchocerciasis are needed to effectively eliminate the infection. The O. volvulus cysteine protease inhibitor (onchocystatin) is an immuno-dominant antigen detected in O. volvulus infections, capable of inducing protective immunity. Here, we explore the onchocystatin for a multi-epitope subunit vaccine candidate targeted against onchocerciasis. A multi-epitope vaccine candidate composed of RS-09 as adjuvant, a CD8⁺ T cell peptide, a CD4⁺ T cell peptide and a B cell peptide concatenated with suitable linkers was computationally constructed. Immune simulation of the vaccine response predicted several aspects of antibody-dependent and cellular-mediated immunity with accompanied B cell and helper T cell immune memory development. The levels of lFN-γ and IL-2 were also predicted to be elevated. Collectively, our results suggest that the multi-epitope vaccine construct has the potential to mimic the natural immunity targeted against onchocerciasis and other related filarial infections, and should be considered for further experimental validations.

     

The intertidal mudflats of Barr Al Hikman,Sultanate of Oman,as feeding,reproduction and nursery grounds for brachyuran crabs

Brachyuran crabs are an important ecological and economical, yet often unstudied aspect of intertidal mudflats of the Arabian Peninsula. Here we provide baseline density estimates of crabs at the relatively pristine intertidal mudflats of Barr Al Hikman (Sultanate of Oman) and provide information on their life cycle and habitat preference. Across the winters of 2012–2015 crabs were sampled on a grid covering the entire intertidal depth gradient. 29 species were found and average densities varied between 12 and 54 crabs/m². Deposit-feeding and herbivorous crabs were the most abundant species across all winters. Size frequency data and the presence of ovigerous females show that most crabs species reproduce in the intertidal area. P. segnis, the most important crab for local fisheries, was found to use the intertidal area as a nursery ground. We analysed the relationships between the two most abundant crab species, Macrophthalmus sulcatus and Thalamita poissonii and the environmental variables: seagrass density, tidal elevation, median grain size and sediment depth using Random Forest models. The predictive capacity of the models and the relative importance of the environmental predictors varied between years, but crab densities in general were positively associated with seagrass density, presumably because seagrass offers feeding habitat.

     

Immunoepidemiological Profiling of Onchocerciasis Patients Reveals Associations with Microfilaria Loads and Ivermectin Intake on Both Individual and Community Levels

Mass drug administration (MDA) programmes against Onchocerca volvulus use ivermectin (IVM) which targets microfilariae (MF), the worm''s offspring. Most infected individuals are hyporesponsive and present regulated immune responses despite high parasite burden. Recently, with MDA programmes, the existence of amicrofilaridermic (a-MF) individuals has become apparent but little is known about their immune responses. Within this immunoepidemiological study, we compared parasitology, pathology and immune profiles in infection-free volunteers and infected individuals that were MF⁺ or a-MF. The latter stemmed from villages in either Central or Ashanti regions of Ghana which, at the time of the study, had received up to eight or only one round of MDA respectively. Interestingly, a-MF patients had fewer nodules and decreased IL-10 responses to all tested stimuli. On the other hand, this patient group displayed contrary IL-5 profiles following in vitro stimulation or in plasma and the dampened response in the latter correlated to reduced eosinophils and associated factors but elevated neutrophils. Furthermore, multivariable regression analysis with covariates MF, IVM or the region (Central vs. Ashanti) revealed that immune responses were associated with different covariates: whereas O. volvulus-specific IL-5 responses were primarily associated with MF, IL-10 secretion had a negative correlation with times of individual IVM therapy (IIT). All plasma parameters (eosinophil cationic protein, IL-5, eosinophils and neutrophils) were highly associated with MF. With regards to IL-17 secretion, although no differences were observed between the groups to filarial-specific or bystander stimuli, these responses were highly associated with the region. These data indicate that immune responses are affected by both, IIT and the rounds of IVM MDA within the community. Consequently, it appears that a lowered infection pressure due to IVM MDA may affect the immune profile of community members even if they have not regularly participated in the programmes.     

Barium variation in Acropora palmata and Montastrea annularis

The barium contents of Acropora palmata and Montastrea annularis are distinct both in their averages (6 and 9 ppm, respectively) and, more impressively, in their variability (5 to 7 ppm and 6 to 15 ppm). A. palmata contained about 50% more Ba²⁺ than M. annularis and exhibited more than three times as much variation, as measured by their respective coefficients of variation. In contrast, the means and coefficients of variation of Sr²⁺ in these groups differed by only 6% and 15%. No obvious environmental or post-depositional causes for the inter-and intra-specific variation of Ba²⁺ could be found. Previous experiments indicate that the partitioning of Ba²⁺ into aragonite depends directly on the rate of precipitation. This suggests that Ba²⁺ does not substitute for Ca²⁺ but instead is incorporated by occlusion. Since growth rates in A. palmata often exceed those of M. annularis, this appears to be the mechanism which generates higher and more variable concentrations of Ba²⁺ in A. palmata.     

Ionization properties of monophosphoinositides in mixed model membranes

Phosphoinositides are found in low concentration in cellular membranes but perform numerous functions such as signaling, membrane trafficking, protein recruitment and modulation of protein activity. Spatiotemporal regulation by enzymes that phosphorylate and dephosphorylate the inositol ring results in the production of seven distinct and functionally diverse derivatives. Ionization properties of the phosphorylated headgroups of anionic lipids have been shown to impact how they interact with proteins and lipids in the membrane. While the ionization properties of the three bis and one tris phosphorylated forms have been studied in physiologically relevant model membranes, that of the monophosphorylated forms (i.e., phosphatidylinositol-3-phosphate (PI3P), phosphatidylinositol-4-phosphate (PI4P), phosphatidylinositol-5-phosphate (PI5P)) has received less attention. Here, we used ³¹P MAS NMR to determine the charge of 5 mol% of the monophosphorylated derivatives in pure dioleoylphosphatidylcholine (DOPC) and DOPC/dioleoylphosphatidylethanolamine (DOPE) bilayers as a function of pH. We find that PI3P, PI4P and PI5P each have unique pKa2 values in a DOPC bilayer, and each is reduced in DOPC/DOPE model membranes through the interaction of their headgroups with DOPE according to the electrostatic-hydrogen bond switch model. In this study, using model membranes mimicking the plasma membrane (inner leaflet), Golgi, nuclear membrane, and endosome (outer leaflet), we show that PI3P, PI4P or PI5P maximize their charge at neutral pH. Our results shed light on the electrostatics of the monophosphorylated headgroups of PI3P, PI4P, and PI5P and form the basis of their intracellular functions.     

Island accessibility and distance from beach influence nesting success of Sooty Falcons Falco concolor in Oman

Colonial island‐breeding birds can be particularly vulnerable to anthropogenic disturbance, which can adversely affect their nesting success. We studied Sooty Falcons Falco concolor breeding on 10 ground‐predator‐free islands in the Sea of Oman during 2007–2014 and evaluated spatio‐temporal trends in the number of breeding pairs occurring on the islands and the factors influencing nesting success. The number of breeding pairs on the islands declined during the study, due mostly to the decline on accessible islands; the rate of decline on islands accessible to humans was double that on inaccessible ones. The number of nests with one or more eggs declined during the study period, and the percentage of nests with eggs that produced one or more chicks showed an increasing trend over time. Sooty Falcon nests located farther away from beaches experienced a significantly higher probability of nesting success than those located closer to beaches. Our results suggest that the number of breeding Sooty Falcons on the islands of northern Oman is declining and that human disturbance may be a contributing factor; this probably mirrors the situation in other parts of the breeding range of this species.     

Hyperreactive Onchocerciasis is Characterized by a Combination of Th17-Th2 Immune Responses and Reduced Regulatory T Cells

Clinical manifestations in onchocerciasis range from generalized onchocerciasis (GEO) to the rare but severe hyperreactive (HO)/sowda form. Since disease pathogenesis is associated with host inflammatory reactions, we investigated whether Th17 responses could be related to aggravated pathology in HO. Using flow cytometry, filarial-specific cytokine responses and PCR arrays, we compared the immune cell profiles, including Th subsets, in individuals presenting the two polar forms of infection and endemic normals (EN). In addition to elevated frequencies of memory CD4⁺ T cells, individuals with HO showed accentuated Th17 and Th2 profiles but decreased CD4⁺CD25^hiFoxp3⁺ regulatory T cells. These profiles included increased IL-17A⁺, IL-4⁺, RORC2⁺ and GATA3⁺CD4⁺ T cell populations. Flow cytometry data was further confirmed using a PCR array since Th17-related genes (IL-17 family members, IL-6, IL-1β and IL-22) and Th2-related (IL-4, IL-13, STAT6) genes were all significantly up-regulated in HO individuals. In addition, stronger Onchocerca volvulus-specific Th2 responses, especially IL-13, were observed in vitro in hyperreactive individuals when compared to GEO or EN groups. This study provides initial evidence that elevated frequencies of Th17 and Th2 cells form part of the immune network instigating the development of severe onchocerciasis.     

Elevated adaptive immune responses are associated with latent infections of Wuchereria bancrofti

In order to guarantee the fulfillment of their complex lifecycle, adult filarial nematodes release millions of microfilariae (MF), which are taken up by mosquito vectors. The current strategy to eliminate lymphatic filariasis as a public health problem focuses upon interrupting this transmission through annual mass drug administration (MDA). It remains unclear however, how many rounds of MDA are required to achieve low enough levels of MF to cease transmission. Interestingly, with the development of further diagnostic tools a relatively neglected cohort of asymptomatic (non-lymphedema) amicrofilaremic (latent) individuals has become apparent. Indeed, epidemiological studies have suggested that there are equal numbers of patent (MF⁺) and latent individuals. Since the latter represent a roadblock for transmission, we studied differences in immune responses of infected asymptomatic male individuals (n = 159) presenting either patent (n = 92 MF⁺) or latent (n = 67 MF⁻) manifestations of Wuchereria bancrofti. These individuals were selected on the basis of MF, circulating filarial antigen in plasma and detectable worm nests. Immunological profiles of either Th1/Th17, Th2, regulatory or innate responses were determined after stimulation of freshly isolated PBMCs with either filarial-specific extract or bystander stimuli. In addition, levels of total and filarial-specific antibodies, both IgG subclasses and IgE, were ascertained from plasma. Results from these individuals were compared with those from 22 healthy volunteers from the same endemic area. Interestingly, we observed that in contrast to MF⁺ patients, latent infected individuals had lower numbers of worm nests and increased adaptive immune responses including antigen-specific IL-5. These data highlight the immunosuppressive status of MF⁺ individuals, regardless of age or clinical hydrocele and reveal immunological profiles associated with latency and immune-mediated suppression of parasite transmission.     

Correction to: The intertidal mudflats of Barr Al Hikman,Sultanate of Oman,as feeding,reproduction and nursery grounds for brachyuran crabs

Hydrobiologia - In the above mentioned publication, incorrect values for P. segnis are shown on the right hand side of Fig. 4. The correct version of Fig. 4 and its caption is...