title: Immunotherapy with mutated onchocystatin fails to enhance the efficacy of a sub-lethal oxytetracycline regimen against Onchocerca ochengi.
abstract: Human onchocerciasis (river blindness), caused by the filarial nematode Onchocerca volvulus, has been successfully controlled by a single drug, ivermectin, for over 25 years. Ivermectin prevents the disease symptoms of severe itching and visual impairment by killing the microfilarial stage, but does not eliminate the adult parasites, necessitating repeated annual treatments. Mass drug administration with ivermectin does not always break transmission in forest zones and is contraindicated in individuals heavily co-infected with Loa loa, while reports of reduced drug efficacy in Ghana and Cameroon may signal the development of resistance. An alternative treatment for onchocerciasis involves targeting the essential Wolbachia symbiont with tetracycline or its derivatives, which are adulticidal. However, implementation of antibiotic therapy has not occurred on a wide scale due to the prolonged treatment regimen required (several weeks). In the bovine Onchocerca ochengi system, it has been shown previously that prolonged oxytetracycline therapy increases eosinophil counts in intradermal nodules, which kill the adult worms by degranulating on their surface. Here, in an "immunochemotherapeutic" approach, we sought to enhance the efficacy of a short, sub-lethal antibiotic regimen against O. ochengi by prior immunotherapy targeting onchocystatin, an immunomodulatory protein located in the adult female worm cuticle. A key asparagine residue in onchocystatin was mutated to ablate immunomodulatory activity, which has been demonstrated previously to markedly improve the protective efficacy of this vaccine candidate when used as an immunoprophylactic. The immunochemotherapeutic regimen was compared with sub-lethal oxytetracycline therapy alone; onchocystatin immunotherapy alone; a gold-standard prolonged, intermittent oxytetracycline regimen; and no treatment (negative control) in naturally infected Cameroonian cattle. Readouts were collected over one year and comprised adult worm viability, dermal microfilarial density, anti-onchocystatin IgG in sera, and eosinophil counts in nodules. Only the gold-standard antibiotic regimen achieved significant killing of adult worms, a profound reduction in microfilarial load, and a sustained increase in local tissue eosinophilia. A small but statistically significant elevation in anti-onchocystatin IgG was observed for several weeks after immunisation in the immunotherapy-only group, but the antibody response in the immunochemotherapy group was more variable. At 12 weeks post-treatment, only a transient and non-significant increase in eosinophil counts was apparent in the immunochemotherapy group. We conclude that the addition of onchocystatin immunotherapy to a sub-lethal antibiotic regimen is insufficient to induce adulticidal activity, although with booster immunisations or the targeting of additional filarial immunomodulatory proteins, the efficacy of this strategy could be strengthened.
authors.affiliation: Institute of Infection & Global Health, University of Liverpool, 146 Brownlow Hill, Liverpool Science Park IC2, Liverpool L3 5RF, UK; Institut de Recherche Agricole pour le Développement, Regional Centre of Wakwa, BP 65 Ngaoundéré, Adamawa Region, Cameroon.
authors.forename: Germanus S
authors.initials: GS
authors.lastname: Bah
authors.affiliation: Institut de Recherche Agricole pour le Développement, Regional Centre of Wakwa, BP 65 Ngaoundéré, Adamawa Region, Cameroon; Cameroon Academy of Sciences, BP 1457 Yaoundé, Centre Region, Cameroon.
authors.forename: Vincent N
authors.initials: VN
authors.lastname: Tanya
authors.affiliation: Institute of Infection & Global Health, University of Liverpool, 146 Brownlow Hill, Liverpool Science Park IC2, Liverpool L3 5RF, UK. Electronic address: blm1@liv.ac.uk.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Makepeace
mesh_terms: D000818:Animals; D002163:Cameroon; D002417:Cattle; D002418:Cattle Diseases / Q000188:drug therapy / Q000628:therapy*; D004359:Drug Therapy, Combination / Q000592:standards / Q000662:veterinary; D005260:Female; D015801:Helminth Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D007074:Immunoglobulin G / Q000097:blood; D007167:Immunotherapy / Q000662:veterinary*; D008297:Male; D009154:Mutation; D009854:Onchocerca / Q000187:drug effects / Q000502:physiology; D010118:Oxytetracycline / Q000494:pharmacology / Q000627:therapeutic use*; D011897:Random Allocation; D016896:Treatment Outcome; D014612:Vaccines / Q000008:administration & dosage / Q000737:chemistry
chemical_list: D015801:Helminth Proteins; D007074:Immunoglobulin G; D014612:Vaccines; C076278:Onchocystatin protein, Onchocerca volvulus; D010118:Oxytetracycline
keywords: Cysteine proteinase inhibitor; Filariasis; Immunochemotherapy; Macrofilaricide; Symbiosis