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1.
Parasitol Int ; 75: 102037, 2020 Apr.
Article in English | MEDLINE | ID: mdl-31841659

ABSTRACT

The aim of this study was to assess the immune response and the protective efficacy elicited by the vaccination with the recombinant Fasciola hepatica myosin regulatory light chain (FhrMRLC) in Adjuplex® adjuvant against the infection with F. hepatica in rats. Four groups of 15 animals each were used for the study, one group was immunized with the recombinant F. hepatica MRLC in Adjuplex® adjuvant and the other groups remained as adjuvant, positive and negative control groups. The parasitological study showed that a statistically significant reduction of 65.1% and 82.1% in fluke burden and fecal egg count, respectively, was detected in vaccinated animals. In addition, vaccination with FhrMRLC induced a well-defined humoral and cellular immune response characterized by a significant production of specific IgG and IL-2, IL-12, TNF-α and IFN-γ; which confirms the immunogenic capacity of the FhrMRLC.


Subject(s)
Fasciola hepatica/physiology , Fascioliasis/immunology , Immunization , Myosin Light Chains/therapeutic use , Th1 Cells/immunology , Animals , Immunity, Cellular , Immunity, Humoral , Male , Myosin Light Chains/immunology , Rats , Rats, Wistar , Recombinant Proteins/immunology , Recombinant Proteins/therapeutic use
2.
Arch Virol ; 161(4): 913-28, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26780774

ABSTRACT

Ectromelia virus (ECTV) is an orthopoxvirus (OPV) that causes mousepox, the murine equivalent of human smallpox. Fas receptor-Fas ligand (FasL) signaling is involved in apoptosis of immune cells and virus-specific cytotoxicity. The Fas/FasL pathway also plays an important role in controlling the local inflammatory response during ECTV infection. Here, the immune response to the ECTV Moscow strain was examined in Fas (-) (lpr), FasL (-) (gld) and C57BL6 wild-type mice. During ECTV-MOS infection, Fas- and FasL mice showed increased viral titers, decreased total numbers of NK cells, CD4(+) and CD8(+) T cells followed by decreased percentages of IFN-γ expressing NK cells, CD4(+) and CD8(+) T cells in spleens and lymph nodes. At day 7 of ECTV-MOS infection, Fas- and FasL-deficient mice had the highest regulatory T cell (Treg) counts in spleen and lymph nodes in contrast to wild-type mice. Furthermore, at days 7 and 10 of the infection, we observed significantly higher numbers of PD-L1-expressing dendritic cells in Fas (-) and FasL (-) mice in comparison to wild-type mice. Experiments in co-cultures of CD4(+) T cells and bone-marrow-derived dendritic cells showed that the lack of bilateral Fas-FasL signalling led to expansion of Tregs. In conclusion, our results demonstrate that during ECTV infection, Fas/FasL can regulate development of tolerogenic DCs and Tregs, leading to an ineffective immune response.


Subject(s)
Ectromelia virus , Ectromelia, Infectious/metabolism , Fas Ligand Protein/metabolism , fas Receptor/metabolism , Animals , Bone Marrow Cells , CD4-Positive T-Lymphocytes/physiology , CD4-Positive T-Lymphocytes/virology , Cell Line , Chlorocebus aethiops , Coculture Techniques , Dendritic Cells/physiology , Dendritic Cells/virology , Ectromelia, Infectious/immunology , Ectromelia, Infectious/virology , Fas Ligand Protein/genetics , Gene Expression Regulation , Killer Cells, Natural , Lymph Nodes , Male , Mice , Signal Transduction , Spleen , Time Factors , fas Receptor/genetics
3.
Mol Biochem Parasitol ; 190(1): 27-37, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23770026

ABSTRACT

Fasciola hepatica infections cause significant global problems in veterinary and human medicine, including causing huge losses in cattle and sheep production. F. hepatica host infection is a multistage process and flukes express papain-like cysteine proteases, termed cathepsins, which play pivotal roles in virulence through host entry, tissue migration and immune evasion. Expression of these proteases is developmentally regulated. Recent studies indicate that excystment of infective larvae is dependent on cysteine proteases and together FhCL3 and FhCB account for over 80% of total protease activity detectable in newly excysted juvenile (NEJ) fluke. This paper focuses on members of the cathepsin L gene family, specifically those belonging to the CL3 clade. The cDNA of two novel cathepsin L3 proteases--FhCL3-1 and FhCL3-2 were cloned. The mRNA transcript expression levels for these enzymes were significantly different at various time points in life development stages obtained in vitro, from dormant metacercariae to NEJ 24h after excystment. Maximum expression levels were observed in NEJ immediately after excystment. In all stages examined by Real Time PCR, FhCL3-2 was expressed at a higher level compared to FhCL3-1 which was expressed only at very low levels. Western blot and immunohistochemical analysis also indicated higher expression of the FhCL3-2 allele and its secretory nature. The ability of antibody responses from rats and sheep challenged with F. hepatica to recognize recombinant FhCL3-1 and FhCL3-2 was shown to differ. Differences were also confirmed through the use of anti-rFhCL3-1 and anti-rFhCL3-2 sera in Western blot analysis of juvenile excretory/secretory (ES) material separated by 2D electrophoresis. These results indicate analysis of relative expression of parasite virulence factors from different populations is required, as this will likely impact the effectiveness of vaccines based on these antigens.


Subject(s)
Cathepsin L/metabolism , Cattle Diseases/parasitology , Fasciola hepatica/enzymology , Fascioliasis/veterinary , Gene Expression Regulation, Enzymologic , Helminth Proteins/metabolism , Sheep Diseases/parasitology , Alleles , Amino Acid Sequence , Animals , Cathepsin L/chemistry , Cathepsin L/genetics , Cattle , Cloning, Molecular , Fasciola hepatica/genetics , Fasciola hepatica/growth & development , Fascioliasis/parasitology , Helminth Proteins/chemistry , Helminth Proteins/genetics , Molecular Sequence Data , Sequence Alignment , Sheep
4.
Pol J Vet Sci ; 12(4): 571-3, 2009.
Article in English | MEDLINE | ID: mdl-20169936

ABSTRACT

The distribution of parasitic nematodes of dogs from three shelters for homeless animals in the Warsaw region (Celestynów and Milanówek near Warsaw, Paluch in Warsaw) was investigated. It was found that since our previous investigations (1993-1995) the prevalence of nematode infections had increased in Celestynów and Milanówek and decreased in the municipal shelter in Warsaw (Paluch). The highest percentage of infected animals was found in Celestynów (as in 1993-1995). What can be the importance of local environmental conditions for the prevalence of nematode infections.


Subject(s)
Dog Diseases/epidemiology , Nematode Infections/veterinary , Animals , Dog Diseases/parasitology , Dogs , Hospitals, Animal , Nematode Infections/epidemiology , Nematode Infections/parasitology , Poland , Prevalence
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