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Biomedical subjects

M Brossard

Publications and source records attributed to M Brossard.

At least 37 records · Page 2Linked to original sources

Investigations on a Swiss area highly endemic for Echinococcus multilocularis.

Anecdotal information suggested that a focus of hyperendemicity may be present in a small area of the Canton of Fribourg in Switzerland. Therefore, the prevalence of E. multilocularis was assessed both in the fox and the rodent population over a two-season-period. A high prevalence ranging between 47% and 56% was consistently determined in the fox population. An Arvicola terrestris population was infected at 39% in the first season of investigation and at 11% in the following season. A subsequent seroepidemiological survey in the population of inhabitants surrounding the area provided no indication of seroconversion and thus no indication of infection for humans. However, a longer-term survey will be needed to assess more precisely the risk of disease occurrence among these inhabitants.

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In vitro production of interleukin-4 and interferon-gamma by lymph node cells from BALB/c mice infested with nymphal Ixodes ricinus ticks.

In this study we compared the ability of lymphocytes taken from axillary and brachial lymph nodes of BALB/c mice that had been infested once three times with 15 nymphal Ixodes ricinus ticks, to produce interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) after in vitro stimulation with concanavalin A (Con A). They released high levels of IL-4 and low levels of IFN-gamma. An increase of IFN-gamma between the first and the third tick infestation was observed. Salivary gland extracts from female I. ricinus ticks induced specific in vitro proliferation of lymphocytes from infested mice. IL-4 production was correlated with the salivary gland extracts' ability to stimulate tick-specific lymphocyte proliferation. Its levels remained high from the first to the third infestation. IFN-gamma production was not necessarily associated with tick salivary gland antigen stimulation. In BALB/c mice, anti-tick immune response induction is regional and the contribution of other similar secondary lymphoid organs is negligible. Only cells from the lymph nodes which drained the tick-fixation site proliferated in vitro in the presence of tick antigens, and when stimulated with Con A produced IL-4 and IFN-gamma.

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IFN-gamma IL-2, and IL-4 mRNA expression in the skin and draining lymph nodes of BALB/c mice repeatedly infested with nymphal Ixodes ricinus ticks.

The skin and draining lymph nodes of BALB/c mice were examined for IFN-gamma, IL-2, and IL-4 mRNA expression by in situ hybridization in three successive infestations with nymphal Ixodes ricinus ticks. IFN-gamma and IL-2 mRNA positive cells were readily detected in lymph node sections during primary antigenic stimulation (72 hr post-tick attachment), whereas hybridization with IL-4 probe yielded no or only a faint positive signal. No changes in the cytokine pattern were observed in lymph node sections from reinfested mice, with IL-4 mRNA always being expressed to a lesser extent than IFN-gamma and IL-2 mRNA. Seventy-two hours post-tick attachment in primary infestation, some infiltrating cells in the skin were positive for IFN-gamma and IL-4 mRNA, but not for IL-2 mRNA. In skin sections of reinfested mice, mRNA coding for IFN-gamma, IL-2, and IL-4 were detected in infiltrating cells. Cells positive for IL-4 mRNA were lower in number than those positive for IFN-gamma and IL-1 mRNA. A significant decrease in the number of IL-4 mRNA positive cells in the tertiary infestation was noted. All together, these results indicate that I. ricinus nymphal ticks antigens are able to elicit expression of IFN-gamma, IL-2 mRNA and to a lesser extent IL-4 mRNA in both skin and draining lymph nodes. In addition, repeated infestations with ticks led to strong expression of IFN-gamma and IL-2 mRNAs in the skin that may be correlated with previous observations showing the occurrence of cutaneous delayed-type hypersensitivity in tick-infested mice. Notably, the cytokine pattern observed in the skin and draining lymph nodes is not associated with a protective immune response in mice against I. ricinus nymphal ticks infestations.

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Effects of human recombinant interleukin-2 on resistance, and on the humoral and cellular response of rabbits infested with adult Ixodes ricinus ticks.

Rabbits were treated with subcutaneous injections of ten doses of 5 x 10(3) units of human recombinant interleukin-2 (IL-2) during a first infestation with five adult pairs Ixodes ricinus per rabbit, while untreated controls were infested by either five (direct control) or 25 pairs (resistant control) per rabbit. During the second infestation with 25 pairs per rabbit in each group, rabbits treated with IL-2 became more resistant than the rabbits in the two untreated control groups. Stronger resistance was manifested by lower engorgement and egg laying weights, and by smaller numbers of normally fed or ovipositioning ticks. IL-2 treatment had no significant effects on the rabbit anti-tick antibody production and the lymphocyte proliferation to a salivary gland extract (SGE). By contrast, the highest cutaneous responses to SGE were observed in the IL-2 treated group. IL-2 may increase rabbit cell-mediated immunity, and stimulate an increase in the production of memory cells during the induction phase of the immune response (first infestation), allowing the development of a strong resistance in lightly infested rabbits.

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Infiltration of CD4+ CD8+ T cells, and expression of ICAM-1, Ia antigens, IL-1 alpha and TNF-alpha in the skin lesion of BALB/c mice undergoing repeated infestations with nymphal Ixodes ricinus ticks.

The skin cellular immune response of BALB/c mice was examined during three successive infestations with nymphal Ixodes ricinus ticks. An immunohistochemical analysis of skin cryostat sections 72 hr post-tick attachment revealed that CD4+ T cells outnumbered CD8+ T cells in all infestations. The CD4+:CD8+ T-cell ratio was 2.2:1 in the primary infestation, then increased to 3.2:1 and 4.7:1 in the secondary and tertiary infestations. No B lymphocytes (CD45R) were detected in the skin of control and infested mice. A positive staining of intercellular adhesion molecule-1 (ICAM-1) on vascular endothelial cells, dendritic cells and some other mononuclear cells was observed in the dermis. Also, a strong positive staining of Ia antigens on dendritic cells and infiltrated mononuclear cells was noted. The staining pattern was more intense and positive cells increased in number in the skin of re-infested mice compared to the primary infestation. In addition, cells such as epidermal keratinocytes, dermal dendritic cells and infiltrated mononuclear cells positive for the 'pro-inflammatory' cytokines interleukin-1 alpha (IL-1 alpha) and tumour necrosis factor-alpha (TNF-alpha) were localized in the skin of infested mice, as detected at the mRNA level by in situ hybridization and at protein level by immunostaining with antibodies. These results suggest that an antigen was presented to infiltrating T lymphocytes which then became activated. This event may explain the cutaneous delayed-type hypersensitivity previously described in tick-infected BALB/c mice. Importantly, this cutaneous reaction was not sufficient to protect the mouse against tick re-infestation. Furthermore, ICAM-1 could mediate, at least in part, the extravasation of inflammatory cells into the skin of infested mice.

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Absence of acquired resistance to nymphal Ixodes ricinus ticks in BALB/c mice developing cutaneous reactions.

BALB/c mice underwent 3 successive infestations with 15 Ixodes ricinus nymphs. No resistance was acquired as assessed by evaluating tick attachment, duration of blood meal, weights of engorged nymphs, and molting success. However, the hosts developed cutaneous immediate- and delayed-type hypersensitivity reactions when reinfested. Histological examination of tick attachment sites showed that inflammatory cells consisting of neutrophils, eosinophils, and mononuclear cells (lymphocytes and monocytes) infiltrated the skin more intensively during reinfestations. The number of intact mast cells did not vary between successive infestations, whereas the number of degranulated mast cells increased in the early stages of reinfestations. Basophils, which represent 12% of total infiltrating cells, were only observed and quantified in the skin of reinfested mice using transmission electron microscopy (TEM). Degranulating eosinophils were also observed by use of TEM.

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Changes in immunity to Ixodes ricinus by rabbits infested at different levels.

Two groups of rabbits were infested twice with different numbers of Ixodes ricinus adults: one group (high infestation) with twenty-five females and twenty-five males and the other group (low infestation) with five pairs. A third infestation was performed in both groups with fifteen adult pairs. Tick biology was monitored for resistance effects. At the second infestation, the feeding and the egg production were more perturbed in ticks fed on high infestation rabbits. The embryogenesis was only affected in ticks from high infestation rabbits. At the third infestation, resistance was increased only in low infestation rabbits, which became more resistant than high infestation rabbits. The blastogenic response of peripheral blood lymphocytes and antibody production against ticks were assessed. A salivary gland extract and an integumental antigen from I. ricinus adult females were able to initiate lymphocyte proliferation. The response was significantly higher in high infestation rabbits, especially at the end of the second infestation, and higher in low infestation rabbits during the third infestation. Non-specific proliferation with concanavalin A was temporarily decreased in both rabbits groups during the first and the second infestations. Specific antibody response to salivary and integument antigens were always the highest in high infestation rabbits. The involvement of tick-induced immunosuppression is discussed.

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Vaccination of cattle against Rhipicephalus appendiculatus with detergent solubilized tick tissue proteins and purified 20 kDa protein.

Three groups of 4 cattle have been vaccinated with either detergent solubilized tick tissue proteins (SMP) of male and female Rhipicephalus appendiculatus, a 20 kDa soluble integumental antigen, a mixture of both SMP and 20 kDa. Two weeks after one booster injection all cattle were challenged by infestation with adult ticks. Treatment had no influence on tick attachment but on cattle vaccinated by the 20 kDa 32.5% fed ticks died (p < 0.001). Moreover, the mean weight of ticks fed on 7 out of 12 vaccinated cattle was significantly lower (p < 0.05 to p < 0.001). Individual differences could be seen where the mean weight reduction was up to 30%. Moreover, ticks fed on 1 (group SMP) or 2 cattle (group 20 kDa) had some difficulties in converting their blood meal into eggs (p < 0.05 to p < 0.001).

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Induction of host resistance to Rhipicephalus appendiculatus in rabbits: effects of immunizing with detergent-solubilized tick tissue proteins.

Resistance to the hard tick, Rhipicephalus appendiculatus, was induced in rabbits by immunizing them with tick tissue proteins extracted with a detergent, Triton X-100. There was 25% mortality in female ticks fed on immunized rabbits as compared with those fed on controls. Similarly, there was a 40% and 60% reduction in the engorged weight and the weight of egg batches, respectively, of ticks fed on immunized rabbits. Western blot analysis of detergent-solubilized tick tissue proteins, carried out using immune sera, recognized a complex pattern of proteins. A strong reaction was observed with proteins with apparent molecular weights of 94,000 and 40,000 daltons.

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Immunity against female Ixodes ricinus L.: effect on feeding and haemoglobin digestion.

Rabbits have been infested 3 times with 10 females and 10 males Ixodes ricinus. Immunity which is acquired when ticks feed on naive rabbits (first infestation) perturbs tick feeding on reinfested animals (third infestation). Then ticks ingest less blood (p less than 0.001). Blood meal digestion in also altered. It was estimated by measuring haemoglobin concentration in ixodid midgut during 20 days after their drop off. After the first infestation this concentration decreases linearly with time (r2(1) = 46.14%, n1 = 63, p less than 0.001). After 3 infestations it is no longer correlated with time, indicating an impaired digestive process (r2(3) = 7.15%, n3 = 49, p greater than 0.05). This observation was corroborated by an analysis of multiple regression. Haemoglobin concentration of tick midgut only correlates with time after a first infestation (r2(1) = 45.25%). In ticks fed on immune animals this concentration is predicted with the quantity of midgut C3 and the weight of fed ticks and not with time (r2(3) = 60.99%).

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Rabbits infested with Ixodes ricinus L. adults: effects of a treatment with cyclosporin A on the biology of ticks fed on naive and immune hosts.

Rabbits have been infested 3 times with 10 female and 10 male Ixodes ricinus. Immunity which is induced when ticks feed on naive animals (1st infestation) perturbs feeding, oviposition and embryogenesis during reinfestations. Treatment of rabbits during a 3rd infestation (resistant animals) with cyclosporin A (CsA), an immunosuppressive agent which works on the cellular compartment (chiefly T helper cells), partially reversed the negative effects of the immunity on the biology of the ticks. Conversely, CsA may also directly affect the reproductive processes of ticks. Thus, the weight of the eggs laid and the egg conversion factor of ticks fed on naive treated hosts (1st infestation) were diminished. In addition, the preoviposition was prolonged, and finally failure in oviposition and hatching occurred more frequently.

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Effects of cyclosporin A on humoral immunity to ticks and on cutaneous immediate and delayed hypersensitivity reactions to Ixodes ricinus L. salivary-gland antigens in re-infested rabbits.

When rabbits are repeatedly infested with Ixodes ricinus L. adults, they acquire resistance to these ticks. A humoral response occurs and the skin of the host becomes progressively sensitized to the saliva of the ectoparasites. The present study examined the effects of cyclosporin A on these two aspects of immunity in re-infested rabbits. Using the enzyme-linked immunosorbent assay (ELISA), moderate suppression of the secondary IgG response to bites of these ticks was observed in animals given this immunosuppressive drug. The agent blocked the immediate cutaneous (type I) reaction normally developed in response to the intradermal injection of I. ricinus salivary gland antigens, as detected by measuring the skinfold thickness. The same method revealed a decreased delayed (type IV) hypersensitivity to these antigens. The present experiments demonstrate the major role of cyclosporin A-sensitive cells (mostly T-lymphocytes, mast cells and basophils) involved in the complex phenomenon of resistance to ticks.

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Repetitive detection by immunoblotting of an integumental 25-kDa antigen in Ixodes ricinus and a corresponding 20-kDa antigen in Rhipicephalus appendiculatus with sera of pluriinfested mice and rabbits.

Mice were pluriinfested with nymphs and rabbits, with adult Ixodes ricinus. As determined by immunoblotting, greater than 50% of sera from these animals reacted against a tick antigen with a molecular weight of 25 kDa, which was detected in total extracts of partially fed I. ricinus females and in tick integumental extract. It was also found in engorged nymphs but was absent from larvae. Sera of I. ricinus-infested rabbits and mice or of rabbits infested with Rhipicephalus appendiculatus adults reacted with a 20-kDa antigen in total extracts of partially fed R. appendiculatus females and the integument of this species.

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C3 levels in the sera of rabbits infested and reinfested with Ixodes ricinus L. and in midguts of fed ticks.

Serum C3 levels of rabbits infested 3 times with Ixodes ricinus L. females and C3 in midgut extracts of fed ticks have been measured by a single radial immunodiffusion test. From the first tick attachment, the mean serum C3 level of hosts increased. A peak of C3 occurred 6 days after the beginning of each infestation and was highest during the third infestation (about 8 times more pronounced than the level measured before the first). After the end of all infestations of the rabbits, the serum C3 level decreased and approached normal levels. Modifications of serum C3 levels during reinfestations influenced C3 contents in the blood meal of fed ticks. Midguts of ticks fed during the third infestation contained more C3 than midguts of ticks of the same engorged weights fed during the first or second infestations.

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Sero-epidemiological survey for alveolar echinococcosis (by Em2-ELISA) of blood donors in an endemic area of Switzerland.

Sera from 17,166 blood donors living in 10 cantons of northern Switzerland in an area endemic for Echinococcus multilocularis were investigated by serological survey for alveolar echinococcosis (AE). A highly species-specific antigen (Em2) and a commonly used E. granulosus hydatid fluid antigen (EgHF) were compared for their suitability in seroepidemiology. EgHF showed a degree of nonspecificity which did not allow direct detection of AE cases. Antibody reaction with Em2 resulted in the detection of 2 asymptomatic clinical cases of AE (seroprevalence 0.01%) within this population of blood donors. A further 4 persons were positive in Em2-ELISA. These 4 persons had negative imaging studies and will be followed serologically and clinically.

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