The elastase, which belongs to the serine protease family, hydrolyses various proteins and may be involved in the parasite invasion. In this study, complete sequence of elastase-1 (TsE) the nematode Trichinella spiralis (Owen, 1835) was cloned into the plasmid pcDNA3.1 as TsE DNA vaccine. After intramuscular vaccination, serum anti-Trichinella antibodies (IgG and subclass IgG1/IgG2a, and IgA), total and specific intestinal mucosal sIgA in mice vaccinated with pcDNA3.1/TsE were measured by ELISA. The results showed that vaccination with pcDNA3.1/TsE induced a systemic humoral immune response (high levels of serum IgG and subclass IgG1/IgG2a and IgA) and local intestinal mucosal immune responses (high levels of TsE-specific sIgA). Vaccination of mice with TsE DNA vaccine also triggered a systemic and local concomitant Th1/Th2 response, as demonstrated by significant elevation of Th1 (IFN-γ and IL-2) / Th2 (IL-4 and IL-10) cytokine levels after the spleen, mesenteric lymph node and Peyer's patch cells from vaccinated mice were stimulated with recombinant TsE (rTsE). The vaccination of mice with pcDNA3.1/TsE displayed a 17% reduction of intestinal adult worms and a 39% reduction of muscle larvae. Our results indicated that TsE DNA vaccine elicited a systemic concomitant Th1/Th2 response and an enteral local sIgA response, and produced a partial protection against infection with T. spiralis. The TsE may be regarded as a potential candidate vaccine target against Trichinella infection. The oral polyvalent vaccines should be developed to improve the protective efficacy of anti-Trichinella vaccines.

• Klíčová slova
• Trichinella, protective immunity, trichinellosis, vaccination,
• MeSH
• DNA vakcíny aplikace a dávkování   MeSH
• myši MeSH
• pankreatická elastasa aplikace a dávkování   imunologie   farmakologie   MeSH
• proteiny červů aplikace a dávkování   imunologie   farmakologie   MeSH
• Trichinella spiralis imunologie   MeSH
• trichinelóza imunologie   parazitologie   MeSH
• vakcinace * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA vakcíny MeSH
• pankreatická elastasa MeSH
• proteiny červů MeSH

The enzyme-linked immunosorbent assay (ELISA) method is recommended for farm surveillance programs and may be useful for epidemiological studies in wildlife or for establishing Trichinella-free areas. In this study, our interest was to compare the specificity and the time of seroconversion of excretory-secretory (E/S) antigens prepared from Trichinella spiralis. A group of eight pigs was inoculated with 500 T. spiralis larvae per animal, and blood sampling was performed at 3 and 4-day intervals during all experiments. The numbers of muscle larvae were determined in four different muscles groups. The larvae per gram burden shows that the most heavily parasitized muscles were the diaphragm [mean = 43.7 larvae per gram (lpg)] and the tongue (mean = 16.9 lpg). Antibody responses were detected by any of eight infected pigs of T. spiralis. Using the ELISA method with E/S antigen, antibodies to T. spiralis were first found on the day 21st p.i. The initial detection of antibodies varied from 21st to 31st day p.i., and the peak was reported 42nd day p.i. Dynamic of antibodies was stable or increased slightly throughout the experimental period (60 days post-inoculation). Our results represent important data for validation of a serological test, especially if blood samples are taken during early stages of infection.

• MeSH
• antigeny helmintové * MeSH
• bránice parazitologie   MeSH
• časové faktory MeSH
• ELISA metody   MeSH
• jazyk parazitologie   MeSH
• nemoci prasat parazitologie   MeSH
• prasata MeSH
• protilátky helmintové krev   MeSH
• sérologické testy MeSH
• svaly parazitologie   MeSH
• Trichinella spiralis imunologie   izolace a purifikace   MeSH
• trichinelóza diagnóza   veterinární   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny helmintové * MeSH
• protilátky helmintové MeSH

Herbivorous animals can play a very important role in spreading trichinellosis. In the study presented here, the susceptibility and distribution of Trichinella spiralis infection was examined in 16 goat kids. The goats were inoculated with 10,000 T. spiralis larvae isolated by artificial digestion methods. The animals were necropsied per two animals in weekly intervals, and the larval burdens in different muscle tissue and anti-Trichinella antibodies measured with the indirect enzyme-linked immunosorbent assay (ELISA) serological method using excretory-secretory (E/S) antigen for detecting anti-Trichinella antibodies were assessed during the experiment. T. spiralis larval burden was maximal at 6 weeks postinoculation (480-5,057 larvae/g according to locality), and the larvae were also found in the myocardium (0.77 larvae/g). In this paper, our next step was to compare the specificity and the time of seroconversion by means of ELISA based on E/S antigen prepared from T. spiralis. Antibody response was detected in all 16 goats. The ELISA test carried out showed the first increments in optical density 2 weeks postinfection (p.i.), reached their peak 4 weeks p.i., and remained elevated from that day until the end of the experiment (10 weeks p.i.). These results indicated that specific anti-Trichinella antibodies in goats persist for a relatively long time.

• MeSH
• ELISA MeSH
• imunoglobulin G krev   MeSH
• kozy MeSH
• larva imunologie   MeSH
• nemoci koz imunologie   parazitologie   MeSH
• protilátky helmintové krev   MeSH
• svaly parazitologie   MeSH
• Trichinella spiralis růst a vývoj   imunologie   izolace a purifikace   MeSH
• trichinelóza imunologie   parazitologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• imunoglobulin G MeSH
• protilátky helmintové MeSH

Susceptibility and reactive manifestation to Trichinella spiralis infection were studied in atypical hosts (sheep) for the period of 247 days. Sheep produced anti-Trichinella antibodies as early on Day 11 (low titer 1:200), with maximum reached at Day 35 (titer 1:800). From Day 42 the antibody level was declining with a negative result of examination on Day 70. Mice exhibited anti-Trichinella antibodies only on Day 32 (titer 1:200). This level was rising, reaching high titer (1:1600) on Day 56. This antibody level persisted until Day 156. In the following period, a rapid decrease in the titer was observed (Graph). On Day 32, T. spiralis larvae in sheep were present in all groups of the muscles examined. The highest larval counts during the entire experiment were detected in the masseter. The initially high counts in the diaphragm and tongue were reduced to only 1/4 or 1/10 at the end of the experiment. In mice, the larvae occurred evenly throughout the entire experiment (Tab. I). The first appearance of a capsule around the T. spiralis larva in muscles was observed on Day 32 p. i. No cell response was detected around the capsule (Fig. 1). Neither was any response observed around necrotizing larvae, even though the surrounding myofibrils were caused to die off (Fig. 2). Certain differences in the degree of myofibril degradation by larvae were evident as early as on Day 32. The least damaged myofibrils were those in the masseter, tongue and diaphragm. This finding correlates with the histological recovery of a different number of necrotized larvae from the individual muscle groups examined. Fresh blood extravassations around larvae were observed on Day 59 (Fig. 3). They could be caused by the migration of larvae to a parasitation site. Live uncapsulated larvae were also found on Day 115 p. i. (Fig. 4). An increased cellular presence around some larvae was observed on Day 84. The larvae surrounded by lymphocytes consequently died off, those without lymphocytic responses formed capsules and survived (Fig. 5). The necrotizing larvae were subject to a powerful phagocytic process, presented by histiocytes, forming multinuclear symplasms (Fig. 6). On Day 11 p. i., larvae inside a capsule were dying off as well. The initial stage of larval necrosis in a capsule is also accompanied by an increased lymphocytic response (Fig. 7). The condition of larvae in capsules and the cellular unresponsiveness as late as on Day 247 indicate the long-lasting viability of the larvae. The capsules surrounding T. spiralis larvae in mice were distinctly seen as early as on Day 32 p. i. Lymphocytic aggregations around the capsule were observed throughout the entire experiment (247 days)-Fig. 8.

• MeSH
• kosterní svaly parazitologie   patologie   MeSH
• myši MeSH
• náchylnost k nemoci MeSH
• nemoci ovcí imunologie   parazitologie   patologie   MeSH
• ovce MeSH
• protilátky helmintové analýza   MeSH
• Trichinella spiralis * imunologie   MeSH
• trichinelóza imunologie   parazitologie   patologie   veterinární   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• protilátky helmintové MeSH

The influence of Trichinella spiralis on infections with Trichuris muris was studied in non-responder B10.BR mice. Mice infected only with T. muris were unable to expel parasites and had many adult worms 35 days later. Infection with 300 larvae of T. spiralis, given seven or 14 (but not 28) days after T. muris, enabled mice to expel up to 90% of T. muris; expulsion of T. spiralis was not altered. Concurrently infected mice produced less T. muris-specific IgG2a antibody than mice infected with T. muris only, and showed higher proliferative responses when spleen and mesenteric lymph node cells were cultured in vitro with T. muris antigens. When T. spiralis was present mucosal mast cells were generated in T. muris-infected mice, whereas almost no mast cells were seen with only T. muris. Lymphocytes from doubly-infected mice produced significantly more interleukin 4 and 5 (IL-4, IL-5) and significantly less interferon-gamma (IFN-gamma) when stimulated in vitro with Concanavalin A (Con-A) than cells from mice infected with T. muris only. These data demonstrate that B10.BR mice, which in single infections produce a Th1 response to T. muris and develop no protective immunity, can mount a protective T-helper-2 (Th2) response and expel T. muris when concurrently infected with the 'Th2-inducing' nematode T. spiralis.

• MeSH
• aktivace lymfocytů imunologie   MeSH
• antigeny helmintové imunologie   MeSH
• cytokiny biosyntéza   MeSH
• imunoglobulin G biosyntéza   MeSH
• inbrední kmeny myší MeSH
• kultivované buňky MeSH
• lymfatické uzliny imunologie   MeSH
• mastocyty imunologie   MeSH
• myši MeSH
• protilátky helmintové biosyntéza   MeSH
• slezina imunologie   MeSH
• T-lymfocyty pomocné-indukující imunologie   MeSH
• Trichinella spiralis imunologie   MeSH
• trichinelóza imunologie   MeSH
• trichurióza imunologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny helmintové MeSH
• cytokiny MeSH
• imunoglobulin G MeSH
• protilátky helmintové MeSH