Protective immunity in mice elicited by living infective third-stage hookworm larvae (Shanghai strain of Ancylostoma caninum). (1/63)

OBJECTIVE: To elucidate the mechanisms of protective immunity in mice elicited by living hookworm (Ancylostoma caninum third-stage infective larvae (L3). METHODS: The number of migrating infective larvae recovered from the lungs was used as an endpoint for vaccine immunity. The timing of maximal L3 lung entry was determined by counting the number of lung larvae at several time points after infection with 500 or 1000 L3. Mice were immunized either orally or subcutaneously with 500 L3, followed by two boosts of L3 once every two weeks. The immunized mice were challenged orally with 500 L3 one week after the final boost. To evaluate the protective immunity, the number of L3 recovered from the lungs of the immunized mice during the time of maximal larval entry was compared with that of controls. Host immunity was also evaluated by comparing circulating anti-L3 antibodies between immunized and controlled mice, using both enzyme immunoassays and immunoblotting techniques, and by lung histopathology. RESULTS: The peak time of larval entry into the lungs occurred 48 hours after infection. Mice immunized either orally or subcutaneously with L3 exhibited a marked reduction (90.2% and 86.2% respectively) in the number of recovered lung larvae in comparison to controls (P < 0.01). The protection might be associated with circulating anti-L3 antibodies, including antibodies directed against 132-200 kDa L3 antigens, as well as three major antigens ranging from 28 to 51 kDa. Larvae migrating through the lungs of vaccinated mice showed cuticular damages accompanied with host-inflammatory cell invasion. CONCLUSIONS: Immunization with living L3 protects mice against lung invasion after challenged with hookworm infection. Vaccine immunity is associated with circulating antibodies against L3 antigens and lung inflammatory responses. The mouse model is potentially useful for developing a hookworm vaccine.  (+info)

Antibody-dependent reductions in mouse hookworm burden after vaccination with Ancylostoma caninum secreted protein 1. (2/63)

Vaccination of mice with either third-stage Ancylostoma caninum infective hookworm larvae (L3) or alum-precipitated recombinant Ancylostoma secreted protein 1 from A. caninum (Ac-ASP-1) results in protection against hookworm challenge infections. Vaccine protection is manifested by reductions in lung hookworm burdens at 48 h postchallenge. Mice actively immunized 4 times with Ac-ASP-1 also exhibited reductions in hookworm burden in the muscles. Hookworm burden reductions from Ac-ASP-1 immunization were associated with elevations in all immunoglobulin subclasses, with the greatest rise observed in host IgG1 and IgG2b. The addition of a fourth immunization resulted in even higher levels of IgG and IgE. In contrast, L3-vaccinated mice exhibited marked elevations in IgG1 and IgM, including anti-Ac-ASP-1 IgM antibody. Passive immunization with pooled sera from recombinant Ac-ASP-1-vaccinated mice also resulted in lung hookworm burden reductions. It is hypothesized that recombinant Ac-ASP-1 vaccinations elicit antibody that interferes with parasite larval migration.  (+info)

Cutaneous larva migrans in travelers: a prospective study, with assessment of therapy with ivermectin. (3/63)

The purpose of this prospective study was to update epidemiological data on cutaneous larva migrans (CLM) and to assess the therapeutic efficacy of ivermectin. We performed the study between June 1994 and December 1998 at our travel clinic. Ivermectin (a single dose of 200 microg/kg) was offered to all the patients with CLM, and its efficacy and tolerability were assessed by a questionnaire. Sixty-four patients were enrolled. All were European and had stayed in tropical areas. After the patients had returned from their destinations, 55% had lesions occur within a mean of 16 days (range, 1-120 days; >1 month in 7 patients). The initial diagnosis was wrong in 55% of patients. The mean number of lesions was 3 (range, 1-15), and the main sites were the feet (48%) and buttocks (23%). The cure rate after a single dose of ivermectin was 77%. In 14 patients, 1 or 2 supplementary doses were necessary, and the overall cure rate was 97%. The median time required for pruritus and lesions to disappear was 3 and 7 days, respectively. No systemic adverse effects were reported. Physicians' knowledge of CLM, which can have a long incubation period, is poor. Single-dose ivermectin therapy appears to be effective and well tolerated, even if several treatments are sometimes necessary.  (+info)

Mitigation of hookworm disease by immunization with soluble extracts of Ancylostoma ceylanicum. (4/63)

Hookworms are a leading cause of anemia in developing countries, and a strategy aimed at reducing pathology caused by blood-feeding adult parasites would be a valuable addition to global control efforts. This article describes experiments designed to induce resistance to the major clinical sequelae (weight loss and anemia) of Ancylostoma ceylanicum hookworm infection in Syrian golden hamsters of the outbred LVG strain. Previously infected animals acquired long-lived resistance to weight loss and anemia caused by a secondary hookworm infection. Furthermore, transfer of pooled serum from twice-infected hamsters to animals undergoing a primary infection was associated with partial resistance to growth delay and anemia. Active vaccination of hamsters with soluble adult hookworm antigens emulsified in alum led to partial protection from hookworm-associated pathology in the absence of reductions in adult worm burden. This intriguing result may have important implications for human vaccine development.  (+info)

Enzyme-linked immunoelectrotransfer blotting analysis of human serologic responses to infective hookworm larval antigen. (5/63)

OBJECTIVE: To explore the possibility of using specific antigens for immunodiagnosis of hookworm disease in endemic area. METHOD: Infective third-stage larvae of the canine hookworm, Ancylostoma caninum (A. caninum), were prepared as the source of antigen. Enzyme-linked immunoelectrotransfer blotting (EITB) was employed as an immunodiagnostic method. RESULTS: Two immunodominant bands of hookworm antigens (42 kDa and 55 kDa) were recognized by the sera of hookworm-infected patients (serum dilution 1:200; antigen centrifuged at 36,000 r/m for 20 minutes, but not by sera from negative controls. CONCLUSION: The 42 kDa and 55 kDa A. caninum antigens might be the specific antigens that could be used for immunodiagnosis of hookworm disease in endemic area.  (+info)

Length of protection by murine vaccination with living infective third-stage hookworm larvae. (6/63)

OBJECTIVE: To determine the length of protection by murine immunization with living third-stage hookworm larvae (L3) as measured by reduction in worm burden and host serologic antibody responses. METHODS: Outbred male (Kunming strain) mice were immunized subcutaneously with 500 L3 once every 2 weeks for a total of immunization for 3 times, and then challenged orally with 1000 L3 for 1 to 8 weeks after the final immunization. Host protective immunity was determined both by the reduction in worm burden as measured by the number of L3 recovered from murine lungs 48-hour post-challenge, as well as by measurement of circulating antibodies. Histopathological responses were also examined. Non-immunized mice served as negative controls. RESULTS: The protection by L3 immunization declined over time. One or 2 weeks after the final immunization, worm burdens were reduced 72% and 77.5% after challenge respectively. In contrast, only 37% reduction in worm burden was observed when the L3 challenge was delayed by 4 weeks and protection was almost entirely lost when there was an 8 week delay between the time of final immunization and challenge. The reduced level of protection over time partially correlated with diminishing L3-specific antibody responses. Host inflammation in the lungs of immunized mice also diminished. CONCLUSION: The protection afforded by living L3 immunization is maximal for the first two weeks after immunization, but then declines significantly over the ensuing weeks.  (+info)

Cutaneous and subcutaneous mast cell and eosinophil responses after challenge in mice vaccinated with living infective third-stage hookworm larvae. (7/63)

OBJECTIVE: To examine the quantitative and qualitative alterations in mast cells and eosinophils distributed in the cutaneous and subcutaneous tissues of hookworm-infected, uninfected and vaccinated mice. METHODS: Outbred male Kunming strain mice, weighing 18-22 g, were vaccinated thrice by subcutaneous inoculation with 500 living third-stage infective larvae (L3) of Ancylostoma caninum (A. caninum) every 2 weeks, and then challenged subcutaneously with 500 L3 one week after the final immunization. Uninfected mice and non-immunized mice but infected with L3 served as controls. The abdominal skin at the site of percutaneous entry was excised from challenged mice at intervals between 6 hours and 7 days after challenge, fixed, and then examined by light microscopy after staining with either toluidine blue or hematoxylin and eosin. RESULTS: The total number of mast cells appearing in cutaneous, and subcutaneous tissues, and underlying abdominal musculature of immunized mice increased significantly compared with non-immunized mice. Mast cells from hookworm-infected mice showed evidence of membrane rupture and degranulation in contrast to the intact appearance of mast cells from uninfected mice. The degree of mast cell degranulation was greater in vaccinated and challenged mice when compared with non-immunized and infected mice. Similarly, eosinophilic infiltration was greatly enhanced after L3 infection. Tissues from vaccinated mice had a greater number of eosinophils than non-immunized mice after infection. CONCLUSIONS: Mast cell alterations appearing earlier than tissue eosinophilic infiltration is major inflammatory response to Ancylostoma caninum (A. caninum) L3 infection in mice. These processes are more obvious in L3-vaccinated mice.  (+info)

Short report: Ancylostoma ceylanicum: exsheathment is not required for successful cryopreservation of third stage hookworm larvae. (8/63)

Third-stage larvae (L3) of the human hookworm parasite Ancylostoma ceylanicum were cultured from the feces of infected hamsters and frozen for up to 100 days in liquid nitrogen. Upon thawing, viable larvae were recovered and used to inoculate naive hamsters. The larvae recovered from this second group of hamsters were used to inoculate a third group of naive animals, which demonstrated that the originally frozen larvae were successfully maintained for two full generations following thawing. These data suggest that exsheathment, which has previously been reported to be essential for successful cryopreservation, is not necessary for recovery of viable, infectious A. ceylanicum L3.  (+info)

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