Gerhard A. Schad

Eosinophils Can Function as Antigen-Presenting Cells To Induce Primary and Secondary Immune Responses to Strongyloides stercoralis

ABSTRACT Several studies have demonstrated roles for eosinophils during innate and adaptive immune responses to helminth infections. However, evidence that eosinophils are capable of initiating an immune response to parasite antigens is lacking. The goal of the present in vitro study was to investigate the potential of eosinophils to serve as antigen-presenting cells (APC) and initiate an immune response to parasite antigens. Purified eosinophils were exposed to soluble Strongyloides stercoralis antigens, and the expression of various surface markers involved in cell activation was examined. Antigen-exposed eosinophils showed a sixfold increase in expression levels of CD69 and major histocompatibility complex (MHC) class II, a fourfold increase in levels of T-cell costimulatory molecule CD86, and a twofold decrease in levels of CD62L compared to eosinophils cultured in medium containing granulocyte-macrophage colony-stimulating factor. The ability of eosinophils to present antigen to T cells was determined by culturing them with T cells in vitro. Eosinophils pulsed with antigen stimulated antigen-specific primed T cells and CD4+ T cells to increase interleukin-5 (IL-5) production. The blocking of MHC class II expression on eosinophils inhibited their ability to induce IL-5 production by CD4+ T cells in culture. Antigen-pulsed eosinophils were able to prime naïve T cells and CD4+ T cells in culture and polarized them into Th2 cells producing IL-5 similar to that induced by antigen-loaded dendritic cells. These results demonstrate that eosinophils are capable of activating antigen-specific Th2 cells inducing the release of cytokines and assist in the priming of naïve T cells to initiate Th2 responses against infection. This study highlights the potential of eosinophils to actively induce immune responses against infection by amplifying antigen-specific Th2-cell responses.

Role of IL-5 in Innate and Adaptive Immunity to Larval Strongyloides stercoralis in Mice

Protective immunity to Strongyloides stercoralis infective larvae in mice has been shown to be dependent on IL-5 based on mAb depletion studies. The goal of this study was to determine the functional role of IL-5 during the innate and adaptive immune response to larval S. stercoralis in mice. In these studies, three strains of mice were used: wild-type C57BL/6J (WT), IL-5 knockout (KO), and IL-5 transgenic (TG). Innate responses to the larvae indicated that there was enhanced survival in the KO animals and decreased survival in the TG animals compared with WT. Furthermore, killing of larvae in TG mice was associated with eosinophil infiltration and degranulation. In studying the adaptive immune response, it was observed that immunization of KO mice did not lead to the development of protective immunity. Experiments were then performed to determine whether KO mice reconstituted with Abs or cells could then develop protective immunity. KO mice displayed protective immunity via a granulocyte-dependent mechanism following injection of purified IgM from immune wild-type animals. Immunity in KO mice could also be reconstituted by the injection of eosinophils at the time of immunization. These eosinophils did not participate in actively killing the challenge infection, but rather were responsible for the induction of a protective Ab response. We conclude that IL-5 is required in the protective immune response for the production of eosinophils, and that eosinophils were involved in larval killing during innate immunity and in the induction of protective Abs in the adaptive immune response.

Hookworm larval infectivity, arrest and amphiparatenesis: the Caenorhabditis elegans daf-c paradigm

Arrested development dramatically alters the life history of some species of soil-transmitted nematodes and elicits profound variations in the epidemiology of the infections they cause. Here, Peter Hotez, John Hawdon and Gerhard Schad show how an understanding of the cellular and molecular bases of arrested development may lead to new approaches for the control of ancylostomiasis and related infections.

Necator americanus and Ancylostoma duodenale: Life history parameters and epidemiological implications of two sympatric hookworms of humans

Abstract The life history data available for two species of hookworm that commonly infect humans, Necator americanus and Ancylostoma duodenale , were compiled. The data were then analyzed in the light of two ecological theories: r and K selection and the constraints of body size on reproduction. Field and laboratory experiments have shown that N. americanus is smaller than A. duodenale and produces fewer but larger eggs. The former is less virulent but the latter is hardier during its free-living stages and can infect orally as well as via the skin. Such species differences reflect the greater opportunism of A. duodenale and the greater degree of accommodation to the human host of N. americanus . A. duodenale optimizes the probability of finding a host and invading it, while N. americanus optimizes the chance of survival once within the host. On the basis of population density, longevity, and fecundity, N. americanus is the more K selected of the two species. However, the theory of r and K selection alone is insufficient to explain the different adaptations of closely related and sympatric parasites with complex life histories. N. americanus does not show greater competitive ability in its freeliving stage, and its mortality is not demonstrably more dependent on density than A. duodenale , as would be predicted by r and K selection theory as presently applied. Epidemiological differences between the two hookworm species are related to such life history parameters as lifespan, fecundity, age-specific survivorship, and density relationships.

Role of Eosinophils and Neutrophils in Innate and Adaptive Protective Immunity to Larval Strongyloides stercoralis in Mice

ABSTRACT The goal of this study was to determine the roles of eosinophils and neutrophils in innate and adaptive protective immunity to larval Strongyloides stercoralis in mice. The experimental approach used was to treat mice with an anti-CCR3 monoclonal antibody to eliminate eosinophils or to use CXCR2−/− mice, which have a severe neutrophil recruitment defect, and then determine the effect of the reduction or elimination of the particular cell type on larval killing. It was determined that eosinophils killed the S. stercoralis larvae in naïve mice, whereas these cells were not required for the accelerated killing of larvae in immunized mice. Experiments using CXCR2−/− mice demonstrated that the reduction in recruitment of neutrophils resulted in significantly reduced innate and adaptive protective immunity. Protective antibody developed in the immunized CXCR2−/− mice, thereby demonstrating that neutrophils were not required for the induction of the adaptive protective immune response. Moreover, transfer of neutrophil-enriched cell populations recovered from either wild-type or CXCR2−/− mice into diffusion chambers containing larvae demonstrated that larval killing occurred with both cell populations when the diffusion chambers were implanted in immunized wild-type mice. Thus, the defect in the CXCR2−/− mice was a defect in the recruitment of the neutrophils and not a defect in the ability of these cells to kill larvae. This study therefore demonstrated that both eosinophils and neutrophils are required in the protective innate immune response, whereas only neutrophils are necessary for the protective adaptive immune response to larval S. stercoralis in mice.

Chemo- and thermosensory neurons: structure and function in animal parasitic nematodes.

Nematode parasites of warm-blooded hosts use chemical and thermal signals in host-finding and in the subsequent resumption of development. The free-living nematode Caenorhabditis elegans is a useful model for investigating the chemo- and thermosensory neurons of such parasites, because the functions of its amphidial neurons are well known from laser microbeam ablation studies. The neurons found in the amphidial channel detect aqueous chemoattractants and repellants; the wing cells-flattened amphidial neurons-detect volatile odorants. The finger cells-digitiform amphidial neurons-are the primary thermoreceptors. Two neuron classes, named ADF and ASI, control entry into the environmentally resistant resting and dispersal dauer larval stage, while the paired ASJ neurons control exit from this stage. Skin-penetrating nematode parasites, i.e. the dog hookworm Ancylostoma caninum, and the threadworm, Strongyloides stercoralis, use thermal and chemical signals for host-finding, while the passively ingested sheep stomach worm, Haemonchus contortus, uses environmental signals to position itself for ingestion. Amphidial neurons presumably recognize these signals. In all species, resumption of development, on entering a host, is probably triggered by host signals also perceived by amphidial neurons. In the amphids of the A. caninum infective larva, there are wing- and finger-cell neurons, as well as neurons ending in cilia-like dendritic processes, some of which presumably recognize a sequence of signals that stimulate these larvae to attach to suitable hosts. The functions of these neurons can be postulated, based on the known functions of their homologs in C. elegans. The threadworm, S. stercoralis, has a complex life cycle. After leaving the host, soil-dwelling larvae may develop either to infective larvae (the life-stage equivalent of dauer larvae) or to free-living adults. As with the dauer larva of C. elegans, two neuron classes control this developmental switch. Amphidial neurons control chemotaxis to a skin extract, and a highly modified amphidial neuron, the lamellar cell, appears to be the primary thermoreceptor, in addition to having chemosensory function. The stomach worm, Haemonchus contortus, depends on ingestion by a grazing host. Once ingested, the infective larva is exposed to profound environmental changes in the rumen. These changes stimulate resumption of development in this species. We hypothesize that resumption of development is under the control of the ASJ neuronal pair. Identification of the neurons that control the infective process could provide the basis for entirely new approaches to parasite control involving interference with development at the time and place of initial host-contact.

Eosinophils Act as Antigen-Presenting Cells to Induce Immunity to Strongyloides stercoralis in Mice

The objective of the present study was to explore the ability of eosinophils to present Strongyloides stercoralis antigen in naive and immunized mice. Antigen-pulsed eosinophils were injected intraperitoneally into naive or immunized mice, and then mice were examined for antigen-specific immune responses. A single inoculation of antigen-pulsed eosinophils was sufficient to prime naive mice and to boost immunized mice for antigen-specific T helper cell type 2 (Th2) immune responses with increased interleukin (IL)-4 and IL-5 production. Mice inoculated 3 times with live eosinophils pulsed with antigen showed significant increases in parasite antigen-specific immunoglobulin (Ig) M and IgG levels in their serum. Antigen-pulsed eosinophils deficient in major histocompatibility complex class II molecules or antigen-pulsed dead eosinophils failed to induce immune responses, thereby demonstrating the requirement for direct interaction between eosinophils and T cells. These experiments demonstrate that eosinophils function as antigen-presenting cells for the induction of the primary and the expansion of the secondary Th2 immune responses to S. stercoralis in mice.

Strongyloides stercoralis: Hyperinfection in immunosuppressed dogs

Hyperinfective strongyloidiasis involving the threadworm , Strongyloides stercoralis, is well known in humans and primates. Although this nematode also frequently parasitizes dogs, canine hyperinfective strongyloidiasis has not been reported. To determine whether a fulminant pattern of nematode development can occur in dogs, and to test the S. stercoralis/dog system for suitability as a model for human hyperinfective and disseminated strongyloidiasis, five canine infections with a dog-derived strain of S. stercoralis were monitored by the quantitative recovery of larvae from feces. Even 3-month-old pups controlled their initial infections successfully, the number of larvae excreted declining to near zero in 90 days. Immunosuppressive treatment with prednisolone, prednisolone and azathiaprine , or niridazole resulted in a rapid return to former or greater intensities of infection, as judged by larval output. Only first stage ( rhabditiform ) larvae were passed in the feces, although third stage (filariform) larvae occurred in the intestinal contents of dogs when they were examined at necropsy. In 3 of the 5 dogs, the adult worm recovery exceeded the inoculated dose greatly and, in one of these, adults and rhabditiform larvae were found in distant, extraintestinal sites. In the remaining 2 of the 5 dogs, the adult worm population was less than the inoculated dose, but, in both, the infection was terminated by the hosts death before hyperinfection could have developed. The observations demonstrate that autoinfection occurs in dogs infected with S. stercoralis and that, if it is allowed to continue for a sufficiently long time in immunosuppressed hosts, massive hyperinfection, and even disseminated infection, may occur. This spectrum of increasingly invasive parasitism closely resembles strongyloidiasis in humans. Therefore, the S. stercoralis/dog system has excellent potential as a model for human hyperinfective and disseminated strongyloidiasis.

IL‐12 eliminates the Th‐2 dependent protective immune response of mice to larval Strongyloides stercoralis

The goal of the present study was to determine if immune‐mediated killing of S. stercoralis L3 in mice could be modulated by shifting from a Th‐2 to a Th‐1 type immune response. L3 killing in immunized mice was ablated in CD4+ T cell‐depleted animals, but not in CD8+ T cell‐depleted or β2‐microglobulin‐deficient mice. Treatment of immunized mice with IL‐4 or IL‐5 neutralizing MoAb significantly reduced the protective effects of vaccination against S. stercoralis, while protective immunity was unimpaired in IFN‐γ knockout mice. Recombinant IL‐12 was administered to infected mice to switch the immune response from a Th‐2 to a Th‐1 type response. Protective immunity was ablated in immunized mice that received IL‐12 therapy. Eosinophil numbers, eosinophil peroxidase levels, and parasite‐specific IgG1 levels were lowered in IL‐12 treated immunized animals, and parasite‐specific IgG2a levels were increased in these animals. The data indicate that eosinophils are important as mediators of larval killing, and that the establishment of Th‐2 type immunity results in killing of infective S. stercoralis L3, while a shift to Th‐1 type immunity abrogates protective responses.

TRICHINELLA SPIRALIS IN AN AGRICULTURAL ECOSYSTEM. II. EVIDENCE FOR NATURAL TRANSMISSION OF TRICHINELLA SPIRALIS SPIRALIS FROM DOMESTIC SWINE TO WILDLIFE

Epidemiological investigations of an outbreak of trichinellosis were carried out in a domestic swine herd and it was established that the parasite also occurred in rats, and in skunks, opossums, and raccoons. Because considerable uncertainty exists regarding the role of sylvatic trichinellosis as a reservoir for the synanthropic cycle, studies were conducted to determine the genetic nature of the various isolates from this ecosystem. Pig infectivity trials, isoenzyme analyses, and repetitive DNA sequence analyses were performed. The results showed that all isolates from the farm environs were genetically similar and that they are related to Trichinella spiralis isolated from domestic pigs. The implication of these findings, in contrast to studies on isolates from wildlife elsewhere, is that this parasite is transmitted from domestic swine to sylvatic hosts and that any control or eradication efforts must take into account the potential for reinfection of hogs from wild animals.