Biology subjects

Hoerauf, A.

Publications and source records attributed to Hoerauf, A..

6 recordsLinked to original sources

bioRxiv2025

Fexinidazole and Corallopyronin A target Wolbachia-infected sheath cells present in filarial nematodes

The discovery of the endosymbiotic bacteria Wolbachia as an obligate symbiont of filarial nematodes has led to antibiotic-based treatments for filarial diseases. While lab and clinical studies have yielded promising results, recent animal studies reveal that Wolbachia levels may rebound following treatment with suboptimal doses of the antibiotic rifampicin. Previous work showed that a likely source of the bacterial rebound in females were dense clusters of Wolbachia in ovarian tissue. The number, size, and density of these Wolbachia clusters were not diminished despite antibiotic treatment. Here we define the cellular characteristics of the Wolbachia clusters in Brugia pahangi (wBp) and identify drugs that also target them. We have evidence that the Wolbachia clusters originate from newly formed sheath cells adjacent to the ovarian Distal Tip Cells. The dramatically enlarged volume of an infected sheath cell is strikingly similar to endosymbiont-induced bacteriocytes found in many insect species. Ultrastructural analysis reveals that the clustered Wolbachia present within the sheath cells exhibit a distinct morphology and form direct connections with the oocyte membrane and possibly the cytoplasm. This includes membrane-based channels providing a connection between Wolbachia-infected sheath cells and oocytes. We also determined that the Wolbachia within the sheath cells are either quiescent or replicating at a very low rate. Screens of known antibiotics and other drugs revealed that two drugs, Fexinidazole and Corallopyronin A, significantly reduced the number of clustered Wolbachia located within the sheath cells.

pathology↗

bioRxiv2023

Eosinophils are an endogenous source of IL-4 during filarial infections and contribute to the development of an optimal T helper 2 response

Interleukin-4 (IL-4) is a central regulator of type 2 immunity, crucial for the defense against multicellular parasites like helminths. This study focuses on its roles and cellular sources during Litomosoides sigmodontis infection, a model for human filarial infections. Our research uncovers eosinophils as a major source of IL-4, especially during the early phase of filarial infection. Using dblGATA mice lacking eosinophil and subsequently eosinophil-derived IL-4, we reveal their profound impact on the Th2 response. Lack of eosinophils impact Th2 polarization and resulted in impaired type 2 cytokine production. Surprisingly, eosinophil deficiency had no impact on macrophage polarization and proliferation as well as on antibody production. These findings shed new light on IL-4 dynamics and eosinophil effector functions in filarial infections. AUTHOR SUMMARYFilarial nematodes can cause severe diseases like onchocerciasis and lymphatic filariasis, posing a significant public health challenge in tropical regions, putting over a billion people at risk. The WHO categorizes these infections as neglected tropical diseases and aims to eliminate onchocerciasis transmission and lymphatic filariasis as a public health issue by 2030. To achieve this goal, we need a better understanding of the protective immune responses involved. Eosinophils have been identified as a key immune cell type in the well-established murine model for filarial infection, Litomosoides sigmodontis. However, their precise roles and interactions with other components of the type 2 immune response remain unclear. Our study reveals that eosinophils play a crucial role as a primary source of interleukin-4, the central cytokine in type 2 immunity. By using dblGATA mice, we found that the absence of eosinophils resulted in a reduced T helper 2 response but did not impact the alternative activation of macrophages or antibody production. In summary, our research uncovers an underappreciated function of eosinophils and their significant influence on type 2 immune responses.

immunology↗

bioRxiv2023

Estimating the number of breeders from helminth larvae with genomic data.

Effective control of helminth infections requires the application of mathematical models to inform control efforts and policy, the development of product profiles for new drugs, and the monitoring of existing drugs for resistance. Key to the success of these approaches is accurately estimating the number of worms within a host, as well as distinguishing, in drug efficacy trials and monitoring, between adults surviving treatment and adults who have reinfected a host following drug treatment. In practice, observing adult worms is often extremely challenging in a patient, as many adult helminths are embedded deep in host tissues. Genetic approaches to infer kinship between larvae or eggs offer a solution to establish adult worm burdens, and to distinguish between treatment failure or treatment success followed by reinfection. Here, we use low-coverage whole-genome, and mitochondrial sequencing, of Onchocerca volvulus larvae to estimate the number of adults contributing to pools of offspring of known and unknown parentage. lcWGS reconstructs full-sibling relationships, resolving the number of unique adult worms contributing to a pool of offspring. Mitochondrial genotyping reconstructs maternal sib-ship, thus estimating the minimum number of adult females within a patient. Further development will improve these techniques for evaluating adult worm burden and trial outcome.

genomics↗

bioRxiv2022

Identification of the onchocerciasis vector in the Kakoi-Koda focus of the Democratic Republic of Congo

BackgroundThe objective of this study was to characterise the vector in a small hyper-endemic focus of onchocerciasis (the Kakoi-Koda focus) which has recently been discovered on the western slopes of the rift valley above Lake Albert. Methodology/Principal FindingsAquatic stages of blackflies were collected by hand from streams and rivers, and anthropophilic adult females were collected by human landing catches. Using a combination of morphotaxonomy and DNA barcoding, the blackflies collected biting humans within the focus were identified as Simulium dentulosum and S. vorax, which were also found breeding in local streams and rivers. Simulium damnosum s.l., S. neavei and S. albivirgulatum were not found (except for a single site in 2009 where crabs were carrying S. neavei). Anthropophilic specimens from the focus were screened for Onchocerca DNA using discriminant qualitative real-time triplex PCR. One specimen of S. vorax was positive for Onchocerca volvulus in the body, and out of 155 S. dentulosum, 30% and 11% were infected and infective (respectively). Conclusions/SignificanceSimulium dentulosum currently appears to be the main vector of human onchocerciasis within the Kakoi-Koda focus, and S. vorax may be a secondary vector. It remains possible that S. neavei was the main (or only) vector in the past having now become rare as a result of the removal of tree-cover and land-use changes. Simulium vorax has previously been shown to support the development of O. volvulus in the laboratory, but this is the first time that S. dentulosum has been implicated as a vector of onchocerciasis, and this raises the possibility that other blackfly species which are not generally considered to be anthropophilic vectors might become vectors under suitable conditions. Because S. dentulosum is not a vector in endemic areas surrounding the Kakoi-Koda focus, it is probable that the Kakoi-Koda focus is significantly isolated. AUTHOR SUMMARYRiver blindness (=onchocerciasis) is a severely debilitating disease caused by the nematode parasite Onchocerca volvulus, and in Africa it is known to be transmitted from person to person by blood-sucking blackflies (Diptera: Simuliidae) of the Simulium damnosum complex, the S. neavei group and (rarely) by S. albivirgulatum. Using classical morphological characteristics and DNA analysis we have unexpectedly identified the vector blackfly as S. dentulosum (and possibly S. vorax) in a small endemic area (the Kakoi-Koda focus) which has been recently discovered on the western slopes of the rift valley in the Democratic Republic of Congo above Lake Albert. In the surrounding endemic areas, the vectors are S. damnosum complex and/or S. neavei (as normally expected), and because S. dentulosum is not a vector in these surrounding areas, it follows that this focus is entomologically isolated from immigrant blackfly species which might otherwise have carried new infections into the Kakoi-Koda focus (and vice-versa). This is of local importance, because it makes elimination of the parasite easier, but our findings have wider significance across Africa, because they raise the possibility that under the right conditions, other common and widely-distributed blackfly species might unexpectedly become man-biters and significant vectors.

ecology↗

bioRxiv2022

Nucleic acid receptor ligands improve vaccination efficacy against the filarial nematode Litomosoides sigmodontis

Infections with helminths affect more than one billion people worldwide. Despite an urgent need there is no vaccine available that would confer long lasting protection against helminth infections. Previous studies indicated that a vaccination with irradiated infective L3 reduces the worm load. This present study investigated whether the additional activation of cytosolic nucleic acid receptors as adjuvant improves the efficacy of a vaccination with irradiated L3 larvae of the rodent filaria Litomosoides sigmodontis. Subcutaneous injection of irradiated L3 larvae in combination with poly(I:C) or 3pRNA resulted in increased neutrophil recruitment to the skin, accompanied by higher IP-10/CXCL10 and IFN-{beta} RNA levels at the site of injection. To investigate the in vivo impact on parasite clearance, BALB/c mice received 3 subcutaneous injections in 2-week intervals with irradiated L3 larvae in combination with poly(I:C) or 3pRNA prior to the challenge infection. Serum analysis before the challenge infection confirmed the induction of L. sigmodontis-specific antibodies in response to the immunization and serum from immunized mice significantly reduced larval motility in vitro with naive cells. 63 days after the challenge infection, vaccination with irradiated L3 larvae in combination with poly(I:C) or 3pRNA led to a significantly greater reduction in adult worm counts by 73% and 57%, respectively, compared to the immunization with irradiated L3 larvae alone (45%). Further, the treatment of L. sigmodontis infection with 3pRNA alone, but not poly(I:C), resulted in a reduced worm burden, supporting the therapeutic potential for the activation of RIG-I with 3pRNA. In conclusion, our data demonstrate that the additional activation of nucleic acid sensing immune receptors boosts the immune response and provides better protection against L. sigmodontis. Thus, the use of nucleic acid receptor agonists as vaccine adjuvants represents a promising novel strategy to improve the efficacy of vaccines against filariae and potentially of other helminths. Author SummaryFilarial nematodes can cause debilitating diseases such as onchocerciasis and lymphatic filariasis that present a major public health burden in the tropics and subtropics, putting more than a billion people at risk of infection. Filarial diseases are transmitted to humans by insect vectors as they take a blood meal. The WHO classifies both filarial infections as neglected tropical diseases and aims to eliminate the transmission of onchocerciasis and eliminate lymphatic filariasis as public health problem by 2030. However, up to date there is no vaccination available that could support the efforts to eliminate filarial diseases and potentially helminth infections in general. Here, we used the well-established murine model for filarial infection, Litomosoides sigmodontis, to test the use of nucleic acid receptor agonists as vaccine adjuvants to enhance local immune responses. We found that infection with L. sigmodontis induces type I IFN and our vaccine strategy enhances the production of type I IFN resulting in increased parasite-specific immune responses and enhanced worm clearance. In summary, our study provides a promising novel approach for a vaccination strategy using cytosolic RNA receptor agonists.

immunology↗

bioRxiv2020

Establishment of an in vitro culture system to study the developmental biology (growth, mating and nodule formation) of Onchocerca volvulus with implications for anti-onchocerca drug discovery and screening

Background Infections with Onchocerca volvulus nematodes remain a threat in Sub-Saharan Africa after two decades of ivermectin mass drug administration. Despite this effort, there is still an urgent need for understanding the parasite biology, especially mating behaviour and nodule formation, as well as development of more potent drugs that can clear the developmental (L3, L4, L5) and adult stages of the parasite and inhibit parasite’s reproductive and behavioural pattern.Methodology/Principal Findings Prior to culture, freshly harvested O. volvulus L3 larvae from dissected Simulium were purified by centrifugation using a 30% Percoll solution to eliminate fly tissue debris and contaminants. Parasites were cultured in both cell-free and cell-based co-culture systems, and monitored daily by microscopic visual inspection. Exhausted culture medium was replenished every 2–3 days. The cell-free culture system supported the viability and motility of O. volvulus larvae for up to 84 days (DMEM–10%NCS), while the co-culture system (DMEM–10%FBS–LLC-MK2) extended the worm survival period to 315 days. Co-culture systems alone promoted the two consecutive parasite moults (L3 to L4 and L4 to L5) with highest moulting rates observed in DMEM–10%FBS–LLC-MK2 (69.2±30 %), while no moult was observed in DMEM–10%NCS–LEC condition. O. volvulus adult worms mating and even mating competitions were observed in DMEM–10% FBS –LLC-MK2 co-culture system. Early nodulogenesis was observed in both DMEM–10% FBS–LLC-MK2 and DMEM– 10%NCS–LLC-MK2 systems.Conclusions/Significance The present study describes an in vitro system in which O. volvulus L3 larvae can be maintained in culture leading to the development of reproductive adult stages. Thus, this platform gives potential for the investigation of mating, mating competition and early stage of nodulogenesis of O. volvulus adult worms that can be used as additional targets for onchocercacidal drug screening.Author summary River blindness affects people living in mostly remote and underserved rural communities in some of the poorest areas of the world. Although significant efforts have been achieved towards the reduction of disease morbidity, onchocerciasis still affect million of people in Sub-Saharan Africa. The current control strategy is the annual mass administration of ivermectin which have accumulated several drawbacks overtime: as the sole microfilaricidal action of the drug, very long treatment period (15-17 years) and reports of ivermectin losing its efficacy; Therefore, raising the urgent need for new onchocercacidal molecules. Our study has established an in vitro platform capable of supporting the growth and development of all developmental stages of O. volvulus (L3 infective stage, L4, L5 and adult worms), moreover the platform provided more insight on O. volvulus adult worms reproductive and behavioural pattern. Our findings provide more avenues for mass production of different parasite stages, the investigation of parasite developmental biology and the identification of targets for drug discovery against different phases of development of this filaria parasiteAbbreviationsAPOCAfrican Programme for Onchocerciasis ControlOCPOnchocerciasis Control ProgrammePTSPost-Treatment SurveillanceHC-04Human Hepatocyte cellsHEK-293embryonic human kidneys cellsLLC-MK2Monkey Kidney cellsLECMouse embryonic lung cellsL3Infective LarvaeL4Stage 4 larvaeL5Stage 5 larvaeRPMIRoswell Park Memorial InstituteDMEMDulbecco’s Minimum Essential MediumMEMMinimum Essential MediumIMDMIscove’s Modified Dulbecco MediumNCTCNew jersey Cell Type CollectionNCSNew-born Calf SerumFBSFoetal Bovine SerumBCSBovine Calf SerumMDAMass Drug AdministrationWHOWorld Health OrganisationCMFLCommunity Microfilariae LoadsEGFEpidermal Growth FactorTGF-αTransforming Growth Factor - alphaIGF - IInsulin-like Growth Factor – IPDGFPalettes-Derived Growth FactorDDTDichlorodiphenyltrichloroethane.View Full Text

developmental biology↗