Fasciola hepatica — the pilot study of in vitro assessing immune response against native and recombinant antigens of the fluke

Abstract

Fasciola hepatica is a liver fluke that infects 2.4 million of people and causes great economical loss in animal production. To date a 100% effective vaccine has not been developed and the disease is controlled by drug therapy. Great efforts are put into development of effective vaccine against parasite what is difficult since Fasciola spp. (like other helmints) during evolutionary process has developed sophisticated and efficient methods to evade immune response. During preliminary experiments it is convenient to use cell lines which are relatively cheap and allow for reproducible comparison of results between laboratories. We stimulated BOMA (bovine monocyte/macrophage cell line) and BOMAC (bovine macrophage cell line) with native or recombinant antigens of Fasciola hepatica and assessed IFN-γ, IL-4 and TNF-α level upon stimulation. We observed diminished secretion of proinflammatory TNF-α in LPS activated BOMA cells stimulated with Excretory/Secretory products of adult fluke (Fh-ES). We also observed greater changes in gene expression in LPS activated BOMA cells than in non activated BOMA cells upon stimulation using Fh-ES. The results show possibility of using cell lines for in vitro research of bovine immune response against liver fluke, although this model still requires validation and further characterization.

Toxocara canis mucins among other excretory-secretory antigens induce in vitro secretion of cytokines by mouse splenocytes

“…P. pastoris has been successfully used for production of many parasitic antigens (Bąska et al 2013a ; Bąska et al 2013b ; Rogé et al 2013 ; Zawistowska-Deniziak et al 2013 ) including T. canis TES-120 (Fong and Lau 2004 ). This expression system is highly effective, secretion of recombinant proteins into the medium simplifies affinity purification and what is the most important Pichia yeast carry out both O - and N -glycosylation (Ahmad et al 2014 ).…”

Fasciola hepatica isolates induce different immune responses in unmaturated bovine macrophages

“…BOMA cell stimulation with Fasciola hepatica excretory – secretory ( Fh -ES) products . The bovine monocyte/macrophage cell line was propagated as previously described (3). Briefly, cells were cultured in Dulbecco’s modified Eagle’s medium (with 5% FCS, 2 mM of glutamine, 100 units/mL of penicillin, 100 μg/mL of streptomycin); during the experiment the cells were seeded into 24-well dishes at a concentration of 6.4 × 10 5 cells/mL and stimulated in quadruplicate with Fh -WeyES or Fh -WildES at a final concentration of 10 μg/mL for 24 h, scraped and centrifuged.…”

The Role of the Intestinal Epithelium in the “Weep and Sweep” Response during Gastro—Intestinal Helminth Infections

“…The high prevalence, detrimental effects, and increasing drug resistance of helminths [ 12 , 13 ] have led to much research on developing vaccines [ 14 , 15 , 16 , 17 , 18 ]; however, despite promising results, there is still much to be done to achieve final success. Fully eradicating helminth infections may even in some aspects be controversial, since they may mitigate autoimmune diseases [ 19 , 20 , 21 ] and allergies [ 22 , 23 ] by dampening Th 1 and inducing Th 2 host immune responses [ 24 , 25 ]. The resulting main effects of this response typically are the presence of alternatively activated macrophages, tissue repair, eosinophilia, and the production of IgE [ 26 , 27 ].…”