concanavalin-a and Tick-Infestations

In the present study we compared the immunological reactions between Rhipicephalus sanguineus tick-infested susceptible (dogs and mice) and tick-resistant hosts (guinea pigs), elucidating some of the components of efficient protective responses against ticks. We found that T-cells from guinea pigs infested with adult ticks proliferate vigorously in the presence of concanavalin A (ConA), whereas ConA-induced cell proliferation of tick-infested mice and dogs was significantly decreased at 43.1 and 94.0%, respectively, compared to non-infested controls. Moreover, cells from mice and dogs submitted to one or three successive infestations did not exhibit a T-cell proliferative response to tick antigens, whilst cells from thrice tick-infested guinea pigs, when cultured with either a tick extract or tick saliva, displayed a significant increase in cell proliferation. Also, we evaluated the response of tick-infested mice to a cutaneous hypersensitivity test induced by a tick extract. Tick-infested mice developed a significant immediate reaction, whereby a 29.9% increase in the footpad thickness was observed. No delayed-type hypersensitivity (DTH) reaction was detected. Finally, the differential cell count at the tick attachment site in repeatedly infested mice exhibited a 6.6- and 4.1-fold increase in the percentage of eosinophils and neutrophils, respectively, compared to non-infested animals, while a decrease of 77.0-40.9 in the percentage of mononuclear cells was observed. The results of the cutaneous hypersensitivity test and the cellular counts at the tick feeding site for mice support the view that tick-infested mice develop an immune response to R. sanguineus ticks very similar to dogs, the natural host of this species of tick, but very different from guinea pigs (resistant host), which develop a DTH reaction in addition to a basophil and mononuclear cell infiltration at the tick-attachment site. In conclusion, saliva introduced during tick infestations reduces the ability of a susceptible animal host to respond to tick antigens that could stimulate a protective immune response. As a consequence, the animals present a lack of DTH response and disturbed cellular migration to tick feeding site, which can represent a deficient response against ticks.

Topics: Animals; Antigens; Cell Division; Concanavalin A; Disease Susceptibility; Dog Diseases; Dogs; Female; Guinea Pigs; Host-Parasite Interactions; Hypersensitivity; Ixodidae; Male; Mice; Mice, Inbred C3H; Saliva; Skin; T-Lymphocytes; Tick Infestations

The effects of repeated infestations with Dermacentor andersoni nymphs on the lymphocyte functions of BALB/c mice were investigated. The in-vitro proliferation responses to concanavalin-A or salivary-gland molecules, the production of cytokines, and the expression of two adhesion molecules-leucocyte function-associated antigen-1 (LFA-1) and very late activation-4 (VLA-4)-were all studied. In addition, the ability of salivary-gland extract or saliva from D. andersoni to modulate expression of lymphocyte adhesion molecules in vitro was determined. The proliferative responses of T-lymphocytes to concanavalin-A were significantly suppressed after first and second infestations, and significant increases in lymphocyte proliferation in the presence of tick salivary-gland antigen were observed in infested mice. After two infestations, production of interleukin-2 was significantly decreased but that of interferon-gamma remained unchanged. Production of interleukin-4 and interleukin-10 was significantly enhanced in infested mice after both the first and second infestations. Expression of LFA-1 and VLA-4 by lymphocytes from infested mice was suppressed. Furthermore, both a salivary-gland extract and the saliva of D. andersoni reduced the in-vitro expression of both of these adhesion molecules by lymphocytes from tick-naive mice.

Topics: Analysis of Variance; Animals; Cell Adhesion Molecules; Cell Division; Cells, Cultured; Concanavalin A; Cytokines; Dermacentor; Female; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Linear Models; Mice; Mice, Inbred BALB C; Recurrence; Salivary Proteins and Peptides; T-Lymphocytes; Tick Infestations

Several studies have revealed that T lymphocytes and cytokines play a crucial role in determining the outcome of parasitic infections in terms of protective immunity. In this study we found that Rhipicephalus sanguineus tick saliva stimulates transforming growth factor-beta (TGF-beta), and reduces interleukin-12 (IL-12) secretion by cells from normal C3H/HeJ mice. Moreover, murine lymph node cells harvested 6 days after the fourth infestation with ticks presented an 82.4% decrease in their proliferative response to concanavalin A (Con A) compared with the response of control cells. In addition, lymph node cells cultured in the presence of Con A showed a T-helper 2-type (Th2-type) cytokine profile, represented by augmented IL-4 and IL-10 and TGF-beta. On the other hand, the IL-2, interferon-gamma (IFN-gamma) and IL-12 synthesis was significantly inhibited. These results indicate that ticks may modulate the host's immune response through saliva injection. Considering that C3H/HeJ mice develop no protective immunity to R. sanguineus infestation, our results suggest that tick-induced Th2-type cytokines and a decreased proliferative response probably lead the host to a susceptible state to both tick and tick-transmitted pathogens.

Topics: Animals; Cell Culture Techniques; Cell Division; Concanavalin A; Cytokines; Dogs; Female; Interleukin-12; Macrophages; Mice; Mice, Inbred C3H; Recurrence; Saliva; Spleen; Th2 Cells; Tick Infestations; Transforming Growth Factor beta

In this study, we investigated tick saliva effects on T cell proliferation, antigen presentation and IFN-gamma-induced macrophage activation, events which are associated with host immune defense mechanisms. Mice repeatedly infested with Rhipicephalus sanguineus ticks, similarly to dogs, did not develop resistance to further infestations. We determined that R. sanguineus tick saliva inhibited, in a dose-dependent manner, both Con-A and specific antigen-induced splenic T cell proliferation. Tick saliva diluted twenty times (64 microg/ml) inhibited Con-A-induced and antigen-specific T cell proliferation in 83% and 69%, respectively. In addition, the inhibition of cell proliferation correlated with a decrease in IL-2 production. Microconcentrator fractionated saliva was tested on a Con-A-induced cell proliferation assay, and showed that one fraction between 3 and 10 kDa and another smaller than 3 kDa can be responsible for the inhibition of T cell proliferation. Although saliva inhibited cell proliferation, it did not impair antigen presentation. Tick saliva further abrogated the killing of intracellular forms of Trypanosoma cruzi by IFN-gamma-activated macrophages. Moreover, saliva-induced macrophage inhibition of IFN-gamma-induced-trypanocidal activity was paralleled with 69% less nitric oxide (NO) production. Finally, tick saliva doubled the production of IL-10 and reduced 84.6% production of IFN-gamma by splenocytes cultured with T. cruzi, suggesting that decreased macrophage NO production may be due to a saliva-induced cytokine imbalance, leading to decreased NO synthase activity. Together, these data indicate that tick saliva can modulate host immune response, thus, contributing to its feeding success and favoring the transmission of tick-borne pathogens.

Topics: Animals; Antigen Presentation; Concanavalin A; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Guinea Pigs; Interferon-gamma; Interleukin-10; Interleukin-2; Lymphocyte Activation; Macrophage Activation; Macrophages; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Nitric Oxide; Saliva; Spleen; T-Lymphocytes; Tick Infestations; Ticks

T cells from BALB/c mice infested 9 days before with Ixodes ricinus nymphs had a suppressed response to in vitro concanavalin A (Con A) stimulation compared to cells from uninfested mice. When laminin (the main component of the extracellular matrix) was used as a coating agent, the Con A response of naive mice was characterized by a decrease in cell proliferation, whereas there was no significant effect on the mitogen response of cells from infested mice. In contrast, an increased response to lipopolysaccharide (LPS) was observed when assaying lymph node cells of infested mice, probably reflecting an increase in B-lymphocyte number or activity. LPS cell stimulation was not modified by laminin. Supernatants of lymph node cells, taken 9 days after the first infestation of mice, stimulated with Con A in vitro, contained interleukin-10 (IL-10) but no significant levels of IL-5 as tested by enzyme-linked immunosorbent assay. At this stage of the infestation all T cells reactive with tick antigens generated in lymph nodes that drain the tick fixation site, were CD4+ cells, as determined by CD4+ depletion. With cells taken 9 days after the third infestation an increase of IL-5 and IL-10 was observed. The IL-10 levels were higher than the IL-5. According to these observations, we conclude that the reduction of T-cell proliferation in response to Con A observed in lymph node cells from infested mice, may be due to the combined effect of laminin interaction with T lymphocytes during migration and IL-10 production by these lymphocytes.

Topics: Animals; Cell Culture Techniques; Concanavalin A; Female; Immune Tolerance; Interleukin-10; Interleukin-5; Ixodes; Laminin; Lipopolysaccharides; Lymph Nodes; Mice; Mice, Inbred BALB C; Salivary Glands; Tick Infestations

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.

Topics: Animals; Axilla; Axillary Vein; Cell Division; Cells, Cultured; Concanavalin A; Female; Interferon-gamma; Interleukin-4; Lymph Nodes; Mice; Mice, Inbred BALB C; T-Lymphocytes; Tick Infestations

The proliferative response of lymphocytes from tick-infested Zebu type, N'Dama and Friesian cattle and acaricide-treated Zebu types and Friesians in concanavalin A (Con A) stimulated cultures was monitored regularly for periods ranging from 11 to 27 months. The numbers of ticks on the animals and the presence of dermatophilosis were also noted. The Friesian cattle carried most and the N'Dama fewest Amblyomma variegatum ticks. The tick-infested Friesians all developed severe clinical dermatophilosis within 5 months of becoming tick-infested. Dermatophilosis lesions on the tick-infested Zebu type and N'Dama cattle were less common and less severe especially in the N'Damas. The proliferative response of lymphocytes from tick-infested Friesians in Con A stimulated cultures fell to almost half that of the acaricide-treated Friesians soon after the former became tick-infested. The tick-infested Zebu types also developed a depressed response compared with the tick-free Zebu group over a 27 month study period. However, the responses of the N'Damas was similar to that of the tick-free Zebu types. The addition of autologous serum to Con A stimulated cultures of lymphocytes derived from the tick-infested Zebu types and N'Damas suppressed their proliferative response compared with that of similar cultures for the tick-free Zebu types.

Topics: Actinomycetales Infections; Animals; Cattle; Cattle Diseases; Cell Division; Cells, Cultured; Concanavalin A; Female; Ghana; In Vitro Techniques; Lymphocyte Activation; Male; Skin Diseases, Bacterial; T-Lymphocytes; Tick Infestations; Ticks

Responses of infested and vaccinated Hereford cattle to Boophilus microplus antigens were measured by enzyme-linked immunosorbent assay (ELISA), lymphocyte blastogenesis assay (LBA) and intradermal skin tests. Responses against soluble salivary gland extracts (SGS), salivary gland membrane (SGM), soluble gut extracts (GS), gut membrane (GM), soluble larval extracts (LS) and larval membrane (LM) antigens were tested. In one experiment, cattle infested with up to 160,000 ticks had positive cellular responses to SGS and significant antibodies against LM, GM, SGM, and SGS. Cellular responses to Concanavalin A were not depressed following infestation. Cattle vaccinated with GM, using Quil A as adjuvant, had positive cellular responses to gut and salivary gland antigens and significant antibody responses to all antigens tested. The antibody levels of vaccinated cattle were significantly higher than the antibody levels of infested cattle (P less than 0.05). In a second experiment, immune responses of cattle infested with 40,000 ticks were studied during 38 days. Cellular responses in LBA to several tick antigens were transiently elevated and significant levels of antibody were measured against LM, GM, SGM and SGS, from day 25 (P less than 0.05). Infested cattle had positive skin reactions following intradermal injection of larval and adult tick antigens (P less than 0.05).

Topics: Animals; Antibody Formation; Antigens; Cattle; Cattle Diseases; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Lymphocyte Activation; Male; Skin Tests; Tick Infestations; Ticks; Vaccination