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Nobel Price 2015 for Dr William C. Campbell

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Present study aimed at analysing the set of epidemiologic, clinical and serological data in appraisal of trichinellosis focus in little towns near Poznań. It was stressed, that parasitic testing of the consumed meat associated with biological appraisal of Trichinella spiralis represent valuable criteria, which are helpful in interpretation mild clinical signs and symptoms, serological data and in establishing management of patients.
The prevalence of Trichinella sp. among 97, 193, 480 swine and 309040 wild boars was determined by artificial digestion method. Pork meat was about 6 times less infected with Trichinella sp. in the years 1997-1998 than in the years 1993-1994. In the years 1993-1994 894 positive Trichinella sp. cases (prevalence - 0,0029%) in pork meat were established. The prevalence of Trichinella larvae infection in pork meat in the years 1997-1998 was 0,00030% - larvae were found in 141 cases, only.
The effect of adriamycin on the behaviour of CD4⁺ and CD8⁺ cells in the course of trichinellosis in mice has been studied. The animals infected with 200 larvae per mouse were administered intraperitoneally adriamycin (Adriblastin from Farmitalia) at 1st and 28th day post infection (dpi), in a dosis of 0.2 mg. The mice were killed weakly for 6 weeks and then at the 60th dpi. The examinations were made on histologic sections from the spleen, mesenteric lymph node, jejunum and masseter muscle using immunofluorescent and immunoenzymatic methods with monoclonal antibody. The mice receiving adriamycin exhibited more CD8⁺ cells in the intestinal mucosa and by the end of the experiment also in the muscles in comparison with the control animals, which, however, did not affect the course of the infection.
The study involved mice of an inbred CFW line from the author’s own culture conducted according to the street light method (Lane Petter and Pearson 1971). All the animals, aged about 3 months and weighing 20 g each, were infected per os with 200 T. spiralis larvae per mouse. A total of 76 mice, divided into two groups, were used. Group I constituted the control and consisted of T. spiralis-infected mice, while Group II consisted of chitosan (chitosan adipinate)-treated mice receiving a dose of 0.4 mg per mouse, administered intra-peritoneally for 20 days (6 days prior to infection until day 13 post-infection). Four mice of each group were sacrificed by decapitation on invasion day 7, 14, 21, 28, 35, 42, and 60. Sections of the jejunum and mandibular muscle were used to prepare populations of cells involved in the inflammatory infiltration. The populations selected were the T (CD4+ and CD8+) lymphocytes and macrophages. The first were identified with immunofluorescence, using labelled monoclonal antibodies, while identification of the latter proceeded immunoenzymatically, with non-labelled monoclonal antibodies. In addition, the infected animals in each group were examined for the presence of parasites: on day 7, 10, 14, and 21 post-infection, the intestinal parasites were counted, while, the muscle-dwelling larvae being enumerated on day 60 post-infection. In this study, the macrophage count in the jejunum mucosa basement membrane of the chitosan-treated mice increased until day 21 post-infection and remained, until the observations were terminated, at a level higher than that in the control. On the other hand, the transversely striated muscles revealed, in addition to T CD4+ and T CD8+ lymphocytes, a stronger macrophage mobilisation throughout the period of observations. The chitosan-treated mouse jejunum were also a site of a faster removal (“expulsion”) of adult parasites than in the control, the muscle larval count in those mice being clearly lower than in the control.
The activity of splenocytes obtained from C3H/w mice during various phases of infection with T. spiralis was assessed using the GvH reaction in irradiated B6C3F₁ recipients of spleen cells. It was found that splenocytes obtained from mice during the intestinal phase of infection (i.e. on day 10 of infection) stimulated the GvH reaction whereas splenocytes obtained during later periods of infection (i.e. on days 20 or 30 of infection) inhibited the GvH reaction. The inhibitory effect persisted until day 60 of infection. Decreased function of T cells in the GvH reaction, as observed during the early muscle phase of trichinellosis (i.e. 30 days post injection of the larvae) was less pronounced in hosts infected with T. spiralis than in those infected with T. pseudospiralis. In addition, significant impairment of the GvH reaction was observed following the mixed transplant of splenocytes obtained from both uninfected C3H/w mice as well as from B6C3F₁ mice infected with T. pseudospiralis. The results indicate that the activity of T lymphocytes in mice infected with T. spiralis changes significantly during different phases of infection.
Lymphocyte-induced angiogenesis (LIA), a vascular response was observed when allogenic immunocompetent lymphocytes of C3H/w mice infected with Trichinella spiralis were inoculated intradermally into irradiated BALB/c. It was shown that intestinal phase of T. spiralis infection stimulated mainly CD5 T cells dependent angiogenesis. The enhancement of angiogenesis did not occur in the early and late muscular phases of the infection. It is suggested, that changes in the intensity of angiogenesis may play a role in spreading of new-born larvae in the host.
Mice infected with 200 Trichinella spiralis larvae were killed at 3-69 days post infection (dpi) and the jejunum and masseter muscles were sectioned in a cryostat and examined in the Tuneli method with "In situ Cell Detected Kid POD" of Boehringer-Mannheim. Data concering the exact localization and dynamie of apoptic cells in both organs are presented. The authors conclude that apoptosis plays a important role in the pathogenesis of trichinellosis.
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