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Biomedical subjects

J Uhlír

Publications and source records attributed to J Uhlír.

At least 19 recordsLinked to original sources

Ultrastructural localization of a sialic acid-specific hemolymph lectin in the hemocytes and other tissues of the hard tick Ixodes ricinus (Acari; Chelicerata).

Lectins have been suggested to function as pattern-recognition molecules in invertebrate immune mechanisms. A lectin from the hemolymph of the tick Ixodes ricinus with main specificity for sialic acid was characterized and antibodies directed against this lectin were prepared. In this study, these antibodies were used to localize the lectin in the tissues of I. ricinus. Immunoreactivity with poly- and monoclonal antibodies was detected in the granules of both types of granular hemocytes, at the membrane of hemocytes, and at the basal laminae surrounding the hemocoel. Furthermore, cells attached to the midgut, invaginations of Géné's organ, and granular inclusions of nephrocytes were labeled. The immunoreactivity detected in hemocytes and the hemocoel lining supports the idea that the hemolymph lectin may function as a recognition molecule in the immune system of I. ricinus. Another function could be protection of eggs that are coated with secretions by Géné's organ. The lectin activity could also be involved in transmission of Borrelia burgdorferi, the causative agent of Lyme borreliosis, and the tick-borne encephalitis virus.

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Novel agglutinin in the midgut of the tick Ixodes ricinus.

Haemagglutination activity (HA) was found and characterized in a midgut homogenate of Ixodes ricinus (L.). HA was induced by tick feeding; it was not detected in starved ticks. In a haemagglutination inhibition test, HA showed an affinity for some carbohydrates (N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, rhamnose, and dulcit) and glycoconjugates (especially lipopolysaccharides). Midgut protein components of 37, 60, 65, and 73 kDa were identified by immunoblotting as potential structural subunits of the new agglutinin.

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Immunosuppression and feeding success of Ixodes ricinus nymphs on BALB/c mice.

The effect of repeated infestations of Ixodes ricinus (L.) nymphs on BALB/c mice was studied. Four successive infectations resulted in an increase of tick feeding success. Tick yield and mean engorged weight increased and the length of the feeding period was reduced significantly (P < 0.05-0.01). The increase of specific anti-tick antibodies was not significant (P > 0.05). The blastogenic response of spleen lymphocytes to T-cell mitogens (Con A and PHA-P) was unimpaired or slightly enhanced, whereas the response to B-cell activators (LPS and PWM) was suppressed, as was the total antibody generation in vitro. The numbers of mast cells in murine skin at the tick attachment sites slightly decreased during the third infestation. The suppression of B-cell competence and of antibody generation, together with decrease of skin mast cell numbers in tick attachment sits, are considered to be responsible for enhancement of tick feeding success.

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Humoral and cellular immune response of BALB/c mice to repeated infestations with Ixodes ricinus nymphs.

The capacity of female BALB/c mice to mount an immune response and effective resistance to repeated infestations with I. ricinus nymphs was studied. An anamnestic antibody response and transient in vitro responsiveness of spleen lymphocytes to tick antigens were demonstrated in repeatedly infested mice. On the other hand, the response to concanavalin A--a T-cell mitogen, was suppressed at the same time. In the presented experiment, BALB/c mice did not manifest tick resistance after three successive infestations (with a reinfestation period of 2 weeks). The possibility of an infestation-dependent modulation of immune response in BALB/c mice is discussed.

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Age-dependent immune response of chickens to feeding by Argas persicus larvae.

Chickens of the dwarf Velsum breed acquired immunity to Argas persicus larvae during repeated infestations with 100 larval ticks in 2-week intervals, when primarily infested at the age of 10 weeks. This immunity was manifested by 74.6% and 75.9% rejection of larvae during secondary and tertiary infestations (P < 0.01), respectively, in relation to controls. The mean weight reductions observed were 16.5% and 12.2% in secondary and tertiary infestations, respectively (P < 0.05). Weak immunity also occurred after secondary and tertiary infestations in chickens infested first at the age of 6 and 8 weeks, but not in chickens aged 2 or 4 weeks at first infestation. No changes were observed in mean feeding periods of larvae engorged on chickens of all age groups (P > 0.05). A significant increase in the leucocyte counts in peripheral blood (P < 0.01 or P < 0.05), mostly of lymphocytes and heterophil granulocytes, followed each infestation in chickens of all age groups. Specific anti-tick circulating antibodies were detected by ELISA in the sera of 10-week-old chickens after primary infestations, in the 4-8-week chicken groups following secondary infestations when aged 6-10 weeks, and in the 2-week chicken group only after tertiary infestations, when the chickens reached the age of 6 weeks. A 53 kDa protein was recognized in the whole-body extract of larval ticks by immune serum during immunoblotting. This was probably the antigenic protein responsible for eliciting the circulating antibodies. The acquired immunity to ticks seen in chickens aged 6 weeks or more may be connected with maturation of the bursa of Fabricius and the development of complete immunocompetence, which is believed to occur in chickens at the age of 4-5 weeks.

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Antigens and glycoproteins of larvae, nymphs and adults of the tick Ixodes ricinus.

Protein components of homogenates of unfed larvae and nymphs of Ixodes ricinus (L.), and of ovary, haemolymph, Malpighian tubules, rectal ampulla, fat body, integument, salivary glands and midgut of partially fed adult females were studied for their antigenicity and carbohydrate moieties using immunoblotting and lectin affinity blotting (LAB) techniques. Comparing the individual anti-larval, anti-nymphal and anti-adult immune sera for their capacity to recognize the specific and trans-stadially cross-reactive antigenic proteins, larval feeding induced the most effective humoral response. The majority of immunogens recognized by rabbit anti-tick immune sera are glycoproteins. Most of the glycosylated antigens were modified with N-type glycans; however, O-type glycans were also demonstrated in some antigens. The correlation of the type of glycosylation with antigenicity, and the sharing of common antigenic epitopes by various tissues, are discussed.

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Effect of repeated infestations of BALB/c mice with Ixodes ricinus nymphs on tick-borne encephalitis virus infection.

The effect of repeated infestations of BALB/c mice with Ixodes ricinus (L.) nymphs on tick borne encephalitis (TBE) virus infection was studied. Enhancement of nymphal feeding, occurring in noninfected mice during the quaternary infestations, was less apparent or absent in female nymphs engorged on TBE virus infected mice. The mice infected with TBE virus during quaternary tick infestation survived significantly longer (P < 0.01) than mice infected with TBE virus during the primary tick infestation. The mean titre of virus in murine blood (determined by plaque assay) was significantly lower (P < 0.01) and the number of nymphs acquiring virus was reduced (P < 0.05) when feeding on hosts infected during the quaternary infestation. The results indicate that repeated infestations of I. ricinus nymphs on BALB/c mice, although enhancing tick feeding, reduced infection with TBE virus when inoculated intraperitoneally.

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Isolation and partial characterisation of an immunogen from the mite, Psoroptes cuniculi.

The immunological reactions of Psoroptes cuniculi protein components with antigen-specific rabbit antisera and with six plant lectins are described. In immunoblotting, the antisera from rabbits heavily and mildly infested with Psoroptes cuniculi recognized from two to five protein components. Affino-blotting analysis showed that eight mite components were glycoproteins with different types of glycan structures. The immunogenic antigen was isolated by both immunoaffinity chromatography and identified by crossed immunoelectrophoresis. It was a glycoprotein with a relative molecular mass of 48 kDa and with complex or high mannose type of oligosaccharides.

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Ivermectin: its effect on the immune system of rabbits and rats infested with ectoparasites.

The influence of subcutaneously administered ivermectin on the specific immune response was studied in rabbits infested with mites (Psoroptes cuniculi) and in rats infested with lice (Polyplax spinulosa). A pronounced specific antibody activity and a change in immunoblotting pattern was observed in rabbits after the ivermectin treatment. However, in rats the antibody activity decreased and the profile of specific antibodies, tested by immunoblotting, remained the same as before the treatment. The specific immune response in rabbits artificially immunized with whole-body Psoroptes cuniculi extract was not affected by ivermectin. It was concluded that ivermectin has no direct effect on the immune response of rabbits and rats and that the enhanced immune response in the mite-infested rabbits was caused by the massive release of antigens associated with the synchronous death of the mites.

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Immunization of rabbits with antigens from Psoroptes cuniculi, the rabbit scab mite.

Rabbits immunized with the whole body extract of Psoroptes cuniculi (Delafond, 1859) developed partial immunity to the infestation with this mite. These rabbits manifested P. cuniculi antigen-induced cell response and a high level of specific serum antibody after the immunization. Electrophoretic separation of the mite extract followed by immunostaining with various sera revealed differences between artificially immunized and naturally infested rabbits to most of the P. cuniculi antigens. However, the specific antibody pattern, that was developed by the immunized rabbits, was not changed after these rabbits were infested with the mites.

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Humoral and cellular immune response of rabbits to Psoroptes cuniculi, the rabbit scab mite.

Rabbits infested for the first time with Psoroptes cuniculi (Group A) and heavily infested ones with unknown aetiology (Group B) were examined for specific serum antibody activity and responsiveness of their peripheral blood lymphocytes by the enzyme-linked immunosorbent assay and lymphocyte transformation assay methods. These parameters were examined after treatment with ivermectin and after subsequent challenge infestation with P. cuniculi. Group A rabbits developed a small number of mite-caused lesions, and exhibited a significant P. cuniculi antigen-induced T cell response and a high level of specific serum antibody. However, both lymphocyte responsiveness and antibody production were observed to be suppressed in Group B rabbits that were highly susceptible to P. cuniculi.

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Effect of ivermectin on the development of serum antibody activity in rabbits infested with Psoroptes cuniculi (Acari: Psoroptidae).

Effect of ivermectin injected subcutaneously in a single dose on the serum specific antibody activity against. Psoroptes cuniculi antigens was studied on four heavily and three weakly P. cuniculi infested rabbits. It has been shown that ivermectin enhanced the production of the specific antibody, especially in weakly infested rabbits. The serum specific antibody activity in both heavily and weakly infested rabbits reached the peak levels on fifth after injection of ivermectin.

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