Trypanosoma cruzi screening in Texas blood donors, 2008-2012.

Chagas disease is an important emerging disease in Texas that results in cardiomyopathy in about 30% of those infected with the parasite Trypanosoma cruzi. Between the years 2008 and 2012, about 1/6500 blood donors were T. cruzi antibody-confirmed positive. We found older persons and minority populations, particularly Hispanic, at highest risk for screening positive for T. cruzi antibodies during routine blood donation. Zip code analysis determined that T. cruzi is associated with poverty. Chagas disease has a significant disease burden and is a cause of substantial economic losses in Texas.

Expression, immunogenicity, histopathology, and potency of a mosquito-based malaria transmission-blocking recombinant vaccine.

Vaccines have been at the forefront of global research efforts to combat malaria, yet despite several vaccine candidates, this goal has yet to be realized. A potentially effective approach to disrupting the spread of malaria is the use of transmission-blocking vaccines (TBV), which prevent the development of malarial parasites within their mosquito vector, thereby abrogating the cascade of secondary infections in humans. Since malaria is transmitted to human hosts by the bite of an obligate insect vector, mosquito species in the genus Anopheles, targeting mosquito midgut antigens that serve as ligands for Plasmodium parasites represents a promising approach to breaking the transmission cycle. The midgut-specific anopheline alanyl aminopeptidase N (AnAPN1) is highly conserved across Anopheles vectors and is a putative ligand for Plasmodium ookinete invasion. We have developed a scalable, high-yield Escherichia coli expression and purification platform for the recombinant AnAPN1 TBV antigen and report on its marked vaccine potency and immunogenicity, its capacity for eliciting transmission-blocking antibodies, and its apparent lack of immunization-associated histopathologies in a small-animal model.

Fusion of Na-ASP-2 with human immunoglobulin Fcγ abrogates histamine release from basophils sensitized with anti-Na-ASP-2 IgE.

Na-ASP-2 is a major protein secreted by infective third-stage larvae (L3) of the human hookworm Necator americanus upon host entry. It was chosen as a lead vaccine candidate for its ability to elicit protective immune responses. However, clinical development of this antigen as a recombinant vaccine was halted because it caused allergic reactions among some of human volunteers previously infected with N. americanus. To prevent IgE-mediated allergic reactions induced by Na-ASP-2 but keep its immunogenicity as a vaccine antigen, we designed and tested a genetically engineered fusion protein, Fcγ/Na-ASP-2, composed of full-length Na-ASP-2 and truncated human IgG Fcγ1 that targets the negative signalling receptor FcγRIIb expressed on pro-allergic cells. The chimeric recombinant Fcγ/Na-ASP-2 protein was expressed in Pichia pastoris and shared the similar antigenicity as native Na-ASP-2. Compared to Na-ASP-2, the chimeric fusion protein efficiently reduced the release of histamine in human basophils sensitized with anti-Na-ASP-2 IgE obtained from individuals living in a hookworm-endemic area. In dogs infected with canine hookworm, Fcγ/Na-ASP-2 resulted in significantly reduced immediate-type skin reactivity when injected intradermally compared with Na-ASP-2. Hamsters vaccinated with Fcγ/Na-ASP-2 formulated with Alhydrogel(®) produced specific IgG that recognized Na-ASP-2 and elicited similar protection level against N. americanus L3 challenge as native Na-ASP-2.

Secretion of a proteolytic anticoagulant by Ancylostoma hookworms.

Hookworms of the genus Ancylostoma secrete an anticoagulant that both inhibits the clotting of human plasma and promotes fibrin clot dissolution. This anticoagulant activity is attributable to a 36,000 dalton proteolytic enzyme. The protease can degrade fibrinogen into five smaller polypeptides that intrinsically have anticoagulating properties, covert plasminogen to a mini-plasminogen-like molecule, and hydrolyze a synthetic peptide substrate with specificity for elastolytic enzymes. It is hypothesized that the parasite uses this enzyme to prevent blood clotting while feeding on villous capillaries.

Global prevalence of latent toxoplasmosis in pregnant women: a systematic review and meta-analysis.

BACKGROUND: Toxoplasma gondii infection, if acquired as an acute infection during pregnancy, can have substantial adverse effects on mothers, fetuses and newborns. Latent toxoplasmosis also causes a variety of pathologies and has been linked to adverse effects on pregnancy. OBJECTIVE: Here, we present results of a comprehensive systematic review and meta-analysis of the global prevalence of latent toxoplasmosis in pregnant women. DATA SOURCE: We searched PubMed, EMBASE, Web of Science, SciELO and Scopus databases for relevant studies that were published between 1 January 1988 and 20 July 2019. STUDY ELIGIBILITY CRITERIA: All population-based, cross-sectional and longitudinal studies reporting the prevalence of latent toxoplasmosis in healthy pregnant women were considered for inclusion. PARTICIPANTS: Pregnant women who were tested for prevalence of latent toxoplasmosis. INTERVENTIONS: There were no interventions. METHOD: We used a random effects model to calculate pooled prevalence estimates with 95% confidence intervals (CIs). We grouped prevalence data according to the geographic regions defined by the World Health Organization (WHO). Multiple subgroup and meta-regression analyses were performed. RESULTS: In total, 311 studies with 320 relevant data sets representing 1 148 677 pregnant women from 91 countries were eligible for inclusion in the meta-analysis. The global prevalence of latent toxoplasmosis in pregnant women was estimated at 33.8% (95% CI, 31.8-35.9%; 345 870/1 148 677). South America had the highest pooled prevalence (56.2%; 50.5-62.8%) of latent toxoplasmosis in pregnant women, whereas the Western Pacific region had the lowest prevalence (11.8%; 8.1-16.0%). A significantly higher prevalence of latent toxoplasmosis was associated with countries with low income and low human development indices (p < 0.001). CONCLUSION: Our results indicate a high level of latent toxoplasmosis in pregnant women, especially in some low- and middle-income countries of Africa and South America, although the local prevalence varied markedly. These results suggest a need for improved prevention and control efforts to reduce the health risks to women and newborns.

A phase 1 study of the safety, reactogenicity, and immunogenicity of a Schistosoma mansoni vaccine with or without glucopyranosyl lipid A aqueous formulation (GLA-AF) in healthy adults from a non-endemic area.

BACKGROUND: Schistosomiasis caused by Schistosoma mansoni (Sm) is a chronic, debilitating and potentially deadly neglected tropical disease. The licensure of a vaccine to prevent schistosomiasis would represent a major breakthrough in public health. METHODS: The safety and immunogenicity of a candidate Sm vaccine were assessed in this phase I, double-blind, dose-escalation trial. Seventy-two healthy Sm-naïve 18-50 year olds were randomized to receive 3 doses ∼ 8 weeks apart of saline placebo, or 10 µg, 30 µg, or 100 µg of recombinant Sm-Tetraspanin-2 vaccine formulated on aluminum hydroxide adjuvant (Sm-TSP-2/Al) with or without 5 µg of glucopyranosyl lipid A aqueous formulation (GLA-AF). Clinical and serologic responses were assessed for 1 year after dose 3. RESULTS: Vaccines were safe and well-tolerated. The most common reactions were injection site tenderness and pain, and headache and fatigue. Tenderness and pain were more frequent in groups receiving vaccine with GLA-AF than placebo (p = 0.0036 and p = 0.0014, respectively). Injection site reactions among those given Sm-TSP-2/Al with GLA-AF lasted 1.22 and 1.33 days longer than those receiving Sm-TSP-2/Al without GLA-AF or placebo (p < 0.001 for both). Dose- and adjuvant-related increases in serum IgG against Sm-TSP-2 were observed. Peak IgG levels occurred 14 days after dose 3. Seroresponse frequencies were low among recipients of Sm-TSP-2/Al without GLA-AF, but higher among subjects receiving 30 µg or 100 µg of Sm-TSP-2/Al with GLA-AF. More seroresponses were observed among those given 30 µg or 100 µg of Sm-TSP-2/Al with GLA-AF compared to placebo (p = 0.023 and p < 0.001, respectively). Seroresponse frequencies were 0%, 30%, 50%, and 89%, respectively, among those given placebo, or 10 µg, 30 µg or 100 µg of Sm-TSP-2/Al with GLA-AF, suggesting a dose-response relationship for Sm-TSP-2/Al with GLA-AF (p = 0.0001). CONCLUSIONS: Sm-TSP-2/Al with or without GLA-AF was safe and well tolerated in a Sm-naïve population. A vaccine like the one under development may represent our best hope to eliminating this neglected tropical disease.

Hookworm: developmental biology of the infectious process.

Hookworms cause severe anemia and malnutrition in developing countries of the tropics, with an estimated one billion people infected worldwide. An in vitro system that models the early events of infection has provided new information about the linkage between the infectious process and the parasite's developmental biology. The cloning and expression of Ancylostoma secreted protein, ASP 1 - a secreted molecule associated with these developmental processes - is an example of how this system allows us to dissect the infectious process at the molecular level.

Modelling heterogeneity and the impact of chemotherapy and vaccination against human hookworm.

There is a growing emphasis on the development of vaccines against helminths (worms), and mathematical models provide a useful tool to assess the impact of new vaccines under a range of scenarios. The present study describes a stochastic individual-based model to assess the relative impact of chemotherapy and vaccination against human hookworm infection and investigates the implications of potential correlations between risk of infection and vaccine efficacy. Vaccination is simulated as a reduction in susceptibility to infection and the model includes population heterogeneities and dynamical waning of protection. To help identify appropriate measures of vaccine impact, we present a novel framework to quantify the vaccine impact on the infection-associated morbidity and introduce a measure of symmetry to study the correspondence between reduction in intensity and reduction in morbidity. Our modelling shows that, in high-transmission settings, the greatest impact of vaccination will be attained when vaccine efficacy is the greatest among individuals harbouring the heaviest worm burdens, and that the decline of morbidity primarily depends on the level of protection attained in the most at risk 8-12% of the population. We also demonstrate that if risk of infection and vaccine protection are correlated, there is not always a direct correspondence between the reduction in worm burden and in morbidity, with the precise relationship varying according to transmission setting.

Ancylostoma secreted protein 2: cloning and characterization of a second member of a family of nematode secreted proteins from Ancylostoma caninum.

Invading infective third-stage larvae (L3) of parasitic nematodes execute a series of programmed developmental events in response to a host-specific signal encountered during infection. One of these early events is the release of excretory/secretory products. Using an in vitro feeding assay that mimics these early events of infection, a protein released by in vitro activated larvae of the hookworm Ancylostoma caninum was identified. This protein, Ac-ASP-2, was partially sequenced, and the cDNA encoding it isolated by PCR and screening of an A. caninum L3 cDNA library. The Ac-asp-2 cDNA encodes a protein of 219 amino acids that is related to a previously identified protein, Ac-ASP-1, from hookworms. Both molecules are members of an evolutionarily diverse family of molecules that include the venom allergens of the Hymenoptera, and the testes specific proteins/sperm-coating glycoproteins of mammals. Homologues are present in nearly all nematodes tested, as demonstrated by PCR-hybridization and database searching. The Ac-asp-2 mRNA is synthesized in all life history stages, but the gene product is released only by L3 activated to feed in vitro. The wide distribution of the Ac-asp-2 in nematodes and its release in response to host specific signals suggests that Ac-ASP-2 serves an important function in nematode physiology and development, and possibly in the infective process of parasitic species.

Ancylostoma caninum anticoagulant peptide: cloning by PCR and expression of soluble, active protein in E. coli.

Ancylostoma caninum Anticoagulant Peptide (AcAP) is the major anticoagulant activity present in extracts of adult Ancylostoma caninum hookworms. This 8.7 kDa protein is a potent and specific inhibitor of human coagulation factor Xa. Using PCR, we have isolated a cDNA encoding for AcAP from an adult A. caninum cDNA library. The 5' end of the AcAP cDNA was identified by reverse transcription PCR (RT-PCR) using A. caninum cDNA and a 5' primer corresponding to a nematode spliced leader sequence. The AcAP cDNA was expressed in E. coli using a prokaryotic expression vector, and the recombinant fusion protein (rAcAP) was purified to homogeneity using nickel resin affinity chromatography and reverse phase HPLC. Purified rAcAP is comparable to the native protein in inhibitor activity, with an apparent equilibrium inhibitory dissociation constant (Ki*) for the inhibition of factor Xa of 265 +/- 71 pM. The purified protein also prolongs the prothrombin and partial thromboplastic times of human plasma in a dose dependent manner.

Weight loss associated with an endotoxin-induced mediator from peritoneal macrophages: the role of cachectin (tumor necrosis factor).

Endotoxin-induced cells of the reticuloendothelial system were shown to produce mediator(s) that evoke a state of cachexia in recipient animals. The factor(s) responsible were assayed in endotoxin-resistant (C3H/HeJ) mice, which were injected with dialyzed conditioned medium obtained from lipopolysaccharide-induced peritoneal macrophages. The mice exhibited weight loss and anorexia, and they died if sufficient quantities of medium were administered. The syndrome was reversible if injections were discontinued. Endotoxin alone did not produce this effect, and no gross pathologic lesions were discernable in the treated animals. In this model system, cachexia appears to result from the action of soluble macromolecules produced by activated macrophages in vitro. Cachectin (murine tumor necrosis factor) is thought to play a central role in this phenomenon.

Cutaneous and subcutaneous granulomata formation in mice immunized and challenged with third-stage infective hookworm (Ancylostoma caninum) larvae.

To determine the inflammatory and immunological mechanisms associated with live third-stage (L3) hookworm larval vaccines, mice were immunized either subcutaneously or orally with three doses of 500 L3 of Ancylostoma caninum at 2-week intervals, and then challenged percutaneously (via abdominal skin) with 500 L3. Non-immunized mice served as negative controls. Skin was excised from post-challenge mice at intervals between 6 h and 28 days, and then examined by light microscopy. In non-immunized mice the L3 exhibited no structural damage and infiltrating inflammatory cells were absent from the surrounding tissues. There were no changes in the cutaneous architecture. In contrast, skin recovered from the immunized mice was edematous and exhibited marked inflammatory changes with resultant destruction of the challenge L3. At 6 h post-challenge the L3 exhibited cuticular swelling and damage; the surrounding tissue was infiltrated by polymorphonuclear inflammatory cells. By 24 h granulomata in the dermis, subcutaneous tissues and underlying abdominal muscles were first observed surrounding dead L3. The number of granulomata peaked at 72 h, with the majority distributed in the subcutaneous tissues. Plasma cells predominated in the early granulomata, but by 3-7 days post-challenge foreign body giant cells began to appear. In some cases, intact and presumably living L3 were noted in the abdominal muscles 14-28 days post-challenge, which suggested that protection against larval challenge was not absolute. Granuloma formation appears to be a major component of the post-vaccination murine host immune response against challenge larvae. The observation generates several hypotheses to investigate the mechanisms of protection afforded by living helminth vaccines.

Electron and light microscopy of peritoneal cellular immune responses in mice vaccinated and challenged with third-stage infective hookworm (Ancylostoma caninum) larvae.

The role of peritoneal macrophages in a murine model of immunity to living hookworm third-stage larvae (L3) was investigated. Mice immunized orally with 500 L3 once every 2 weeks for three times were challenged intraperitoneally with 2000 L3 1 week after the final immunization. The challenged larvae were collected from the peritoneal cavity at intervals between 2 and 72 h and then examined by inverted light microscopy, scanning electron microscopy and transmission electron microscopy. Peritoneal cellular responses in non-immunized mice served as negative controls. The numbers of peritoneal macrophages in immunized mice were 6-7-fold higher than in non-immunized mice. In the peritoneal cavity of immunized mice, host macrophages adhered to the challenged L3 within 2 h and created a cocoon-like encasing which surrounded the parasite. Extensive damage to the L3 was observed which included swelling, collapse and deformation of the larval cuticle. Lysis and vacuolization of the parasite's internal structures were also observed. In contrast, no significant cellular adherence and damage were observed in L3 recovered from non-immunized mice. L3-specific antibody levels were also elevated in the peritoneum of immunized mice relative to non-immunized controls. These studies implicate macrophages as important effector cells in hookworm larval vaccine immunity.

Epidemiology of human hookworm infections among adult villagers in Hejiang and Santai Counties, Sichuan Province, China.

Hookworm infection as well as other intestinal nematodiases are endemic to Sichuan Province in China. In order to research the prevalence and intensity of these infections we visited two villages in Hejiang County (southern Sichuan Province) and Santai County (northwestern Sichuan Province) between July and October of 1997. Fecal examinations were performed on adult villagers over the age of 15 years (currently children under this age are dewormed annually with anthelmintic drugs). Among 310 residents of Lugao Village (Hejiang County), 87, 63 and 60% were infected with hookworm, Ascaris or Trichuris, respectively. The prevalence of hookworm determined to rise linearly with age (r = 0.97). High intensity infections with hookworm still occur in this region as 22% of the residents have over 3000 eggs per gram (PEG) of feces as determined by quantitative egg counts. The majority of these individuals harbored mixed infection with Necator americanus and Ancylostoma duodenale, although the former predominated when adult hookworms were collected from 30 village residents treated with pyrantel pamoate. In contrast, among the 334 Xinjian villagers examined (Santai County) the majority harbored predominantly light hookworm infections--66.1% of the residents has less than 400 EPG by quantitative fecal examination and only 3.7% exhibited greater than 3000 EPG. Again, N. americanus was the predominant hookworm seen after worm expulsion. We have round that despite economic development which is occurring in some parts of China, significant hookworm infections and clinical hookworm anemia still exist in areas of Sichuan Province. In Hejiang County we found that the intensity of hookworm infection has actually risen within the last 10 years. Hookworm is a medical problem among the elderly in Sichuan.

Electron and light microscopy of neutrophil responses in mice vaccinated and challenged with third-stage infective hookworm (Ancylostoma caninum) larvae.

The role of neutrophils in mediating host inflammation was examined in mice vaccinated with living third-stage infective hookworm larvae (L3). Mice were vaccinated by oral immunization with 500 L3 (Ancylostoma caninum) once every 2 weeks for a total of three immunizations. The vaccinated mice were then challenged intraperitoneally with 2000 L3) 1 week after the final immunization. To stimulate peritoneal production of neutrophils, 2 ml of 2% glycogen were injected intraperitoneally at 16 h prior to the challenge infection. Neutrophils were found to comprise 85% of the peritoneal cell population. L3 from the challenge infection were collected and then examined at timed intervals by inverted light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Greater than a fivefold increase in the total numbers of peritoneal cells was noted in the vaccinated mice as compared to unvaccinated mice. In the peritoneal cavity of vaccinated mice, the neutrophils adhered to the L3 within 2 h, and over 55% of the L3 were surround by clusters of neutrophils to form a sausage-like sheath 4 h later. At 24-72 h after challenge, almost all of the L3 recovered from the vaccinated mice were covered with thick clusters of cells. Both SEM and TEM demonstrated extensive ultrastructural damage to the L3. In contrast, the L3 recovered from the unvaccinated mice appeared to be unaffected by neutrophils. These studies suggest that neutrophils, like macrophages, can have an important role as effector cells in L3-vaccinated mice.

Epidemiology of Necator americanus hookworm infections in Xiulongkan Village, Hainan Province, China: high prevalence and intensity among middle-aged and elderly residents.

Hookworm is highly endemic to Hainan Province, an island located in the South China Sea. To investigate the prevalence and intensity of infection in the area, the village of Xiulongkan was surveyed between April and July 1998. A cross-sectional study was conducted in which fecal samples of 80% of the village residents (631 individuals) were tested for the presence of helminth eggs. Hookworm was the predominant intestinal helminth in Xiulongkan, where it was determined that 60% of those tested were infected. Necator americanus was the predominant species of hookworm in this population. The prevalence of hookworm increased with age, and then leveled to a plateau for ages 41 yr and up. This observation was in contrast to infections with Ascaris lumbricoides, where the highest prevalences occurred among school-aged children. Women had a significantly higher prevalence of hookworm than men and this difference emerged in early adulthood. The intensity of hookworm infection also significantly increased with age, with the highest intensity infections occurring among middle-aged and elderly residents. Females were more likely to have moderate or heavy infections, whereas males were more likely to have light infections. The rates of hookworm transmission are particularly high among the middle-aged and elderly residents of Xiulongkan.

Hookworm infection.

It retards growth and intellectual development in millions of children yet is largely ignored by researchers. New findings suggest excellent possibilities for a vaccine.

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

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.

Length of protection by murine vaccination with living infective third-stage hookworm larvae.

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.

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

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.