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Establishment of Babesia laboratory model and its experimental application
JALOVECKÁ, Marie
Growing incidence of infections caused by the tick-transmitted protozoan parasite Babesia spp. defines babesiosis as an emerging disease from the aspect of human and veterinary medicine. The thesis provides an insight to biology of two main agents of human babesiosis, Babesia microti and Babesia divergens. We introduce here the fully optimized quantification model of Babesia parasite enabling the detailed investigation of the parasite developmental cycle and identification of molecules playing a role in its acquisition and transmission by the vector Ixodes ricinus. Novel and detailed information about Babesia dissemination within the tick tissues are given by newly implemented visualization and quantification techniques. Special emphasis is paid to parasite development in the tick salivary glands, the primary site responsible for parasite transmission from the vector into the host. Using gene-specific silencing we screene the tick immune pathways including effector molecules and evaluate their role in Babesia acquisition. We also provide a detailed view to Babesia parasite sexual commitment by monitoring its kinetics upon various stimuli. Moreover, a new direction of anti-babesial therapy is proposed by validation of the Babesia proteasome as a drug target. Overall, the research presented in the thesis extends the current knowledge of the Babesia parasite biology including molecular interactions at the tick-Babesia interface and thereby could significantly contribute to a potential control of babesiosis.
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The immune response of mice to gastric cryptosporidium infections
JALOVECKÁ, Marie
The immune response in stomach mucosa during the primary infection and re-infection of Cryptosporidium muris (TS03 and CB03) and C. andersoni in immunocompetent BALB/c mice was observed in this study. No significant differences in the induction of a cellular response were observed in mice infected with the two strains of C. muris. Significantly elevated migration of T-lymphocytes (more than 1000-fold), especially CD8+ T lymphocytes, to the stomach mucosa was described during primary infection. Moreover, the persisting severalfold increased level of T-lymphocytes in stomach epithelium was observed 2 months after recovery from the primary cryptosporidiosis. Very low level of IFN-{$\gamma$} production in ex vivo cultures of splenocytes was recorded during the course of the primary infection (0.5 ng/ml), whereas during reinfection the concentration of IFN-{$\gamma$} rapidly increased 22-fold (10.7 ng/ml). After infection of BALB/c mice with C. andersoni LI03, migration of T-lymphocytes and production of INF-{$\gamma$} in ex vivo splenocyte primary cultures was also observed, even though this isolate of C. andersoni does not infect Mus musculus. These results imply that the CD8+ T-lymphocytes are involved in the immune response to gastric cryptosporidiosis and could play an important role in the elimination of C. muris infection in mice.
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Development of protective immune response in gastric mucosa of mice infected with \kur{Cryptosporidium muris} and \kur{Cryptosporidium andersoni}
JALOVECKÁ, Marie
The development of immune response accountable for the ability to control Cryptosporidium muris TS03 infection was studied using immunocompetent and various types of immunodeficient mouse models. Subsequently the immune response was characterized by analysis of leukocyte infiltration and cytokine production in gastric epithelium. Moreover, the potentiality of immunocompetent mice to develop effective immune response to C. andersoni LI03 infection with consequent protection to consequent infection of the same mice with C. muris TS03 was also studied by monitoring oocysts shedding, leukocyte infiltration of the gastric mucosa and cytokine production in ex vivo cultures of splenocytes.
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The immune response of mice to gastric cryptosporidium infections
JALOVECKÁ, Marie
The immune response in stomach mucosa during the primary infection and re-infection of Cryptosporidium muris (TS03 and CB03) and C. andersoni in immunocompetent BALB/c mice was observed in this study. No significant differences in the induction of a cellular response were observed in mice infected with the two strains of C. muris. Significantly elevated migration of T-lymphocytes (more than 1000-fold), especially CD8+ T lymphocytes, to the stomach mucosa was described during primary infection. Moreover, the persisting severalfold increased level of T-lymphocytes in stomach epithelium was observed 2 months after recovery from the primary cryptosporidiosis. Very low level of IFN-{$\gamma$} production in ex vivo cultures of splenocytes was recorded during the course of the primary infection (0.5 ng/ml), whereas during reinfection the concentration of IFN-{$\gamma$} rapidly increased 22-fold (10.7 ng/ml). After infection of BALB/c mice with C. andersoni LI03, migration of T-lymphocytes and production of INF-{$\gamma$} in ex vivo splenocyte primary cultures was also observed, even though this isolate of C. andersoni does not infect Mus musculus. These results imply that the CD8+ T-lymphocytes are involved in the immune response to gastric cryptosporidiosis and could play an important role in the elimination of C. muris infection in mice.
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