Kinetics Of Expulsion Research Articles

Mast cell deficiency in mice results in biomass overgrowth and delayed expulsion of the rat tapeworm Hymenolepis diminuta.

Infection with helminth parasites evokes a complex cellular response in the host, where granulocytes (i.e. eosinophils, basophils and mast cells (MCs)) feature prominently. In addition to being used as markers of helminthic infections, MCs have been implicated in worm expulsion since animals defective in c-kit signaling, which results in diminished MC numbers, can have delayed worm expulsion. The role of MCs in the rejection of the rat tapeworm, Hymenolepsis diminuta, from the non-permissive mouse host is not known. MC-deficient mice display a delay in the expulsion of H. diminuta that is accompanied by a less intense splenic Th2 response, as determined by in vitro release of interleukin (IL)-4, IL-5 and IL-13 cytokines. Moreover, worms retrieved from MC-deficient mice were larger than those from wild-type (WT) mice. Assessment of gut-derived IL-25, IL-33, thymic stromal lymphopoietin revealed lower levels in uninfected MC-deficient mice compared with WT, suggesting a role for MCs in homeostatic control of these cytokines: differences in these gut cytokines between the mouse strains were not observed after infection with H. diminuta. Finally, mice infected with H. diminuta display less severe dinitrobenzene sulphonic acid (DNBS)-induced colitis, and this beneficial effect of the worm was unaltered in MC-deficient mice challenged with DNBS, as assessed by a macroscopic disease score. Thus, while MCs are not essential for rejection of H. diminuta from mice, their absence slows the kinetics of expulsion allowing the development of greater worm biomass prior to successful rejection of the parasitic burden.

The circadian regulator BMAL1 programmes responses to parasitic worm infection via a dendritic cell clock

Resistance to the intestinal parasitic helminth Trichuris muris requires T-helper 2 (TH2) cellular and associated IgG1 responses, with expulsion typically taking up to 4 weeks in mice. Here, we show that the time-of-day of the initial infection affects efficiency of worm expulsion, with strong TH2 bias and early expulsion in morning-infected mice. Conversely, mice infected at the start of the night show delayed resistance to infection, and this is associated with feeding-driven metabolic cues, such that feeding restriction to the day-time in normally nocturnal-feeding mice disrupts parasitic expulsion kinetics. We deleted the circadian regulator BMAL1 in antigen-presenting dendritic cells (DCs) in vivo and found a loss of time-of-day dependency of helminth expulsion. RNAseq analyses revealed that IL-12 responses to worm antigen by circadian-synchronised DCs were dependent on BMAL1. Therefore, we find that circadian machinery in DCs contributes to the TH1/TH2 balance, and that environmental, or genetic perturbation of the DC clock results in altered parasite expulsion kinetics.

Disruption of Th2 Immunity Results in a Gender-Specific Expansion of IL-13 Producing Accessory NK Cells during Helminth Infection

Host gender has previously been identified as a determining factor in the resolution of Trichuris muris infection in mice lacking IL-4 (IL-4KO BALB/c). Worm expulsion in these mice is delayed, but occurs in females. In this study we were able to demonstrate delayed expulsion occurs at day 26 post infection and is associated with the production of the key Th2-associated cytokine IL-13 by both CD4(+) T cells and an auxiliary DX5(+) NK cell source, as well as a concurrent reduction in proinflammatory cytokines. NK cell number was comparably increased in both sexes, but NK cells from male mice were found to express higher levels of the chemokine receptor CXCR3. Depletion of CD4(+) T cells completely prevented parasite expulsion, whereas loss of NK cells resulted in a mild, but significant delay. Furthermore, IL-18 is a cytokine with the capacity to enhance both Th1 and Th2 responses found to be dispensable for worm expulsion in female mice but was a key factor for the suppression of the Th2 response in male IL-4KO mice. In contrast neutralization of IFN-gamma resulted in a complete restoration of typical wild-type BALB/c expulsion kinetics. This study sheds further light on the role of accessory NK cells in supplementing the IL-13-driven immune response when normal Th2 immunity is disrupted, and further identifies host gender as a key factor in determining the generation of "NK cell help".

Expulsion of Trichuris muris is associated with increased expression of angiogenin 4 in the gut and increased acidity of mucins within the goblet cell

BackgroundTrichuris muris in the mouse is an invaluable model for infection of man with the gastrointestinal nematode Trichuris trichiura. Three T. muris isolates have been studied, the Edinburgh (E), the Japan (J) and the Sobreda (S) isolates. The S isolate survives to chronicity within the C57BL/6 host whereas E and J are expelled prior to reaching fecundity. How the S isolate survives so successfully in its host is unclear.ResultsMicroarray analysis was used as a tool to identify genes whose expression could determine the differences in expulsion kinetics between the E and S T. muris isolates. Clear differences in gene expression profiles were evident as early as day 7 post-infection (p.i.). 43 probe sets associated with immune and defence responses were up-regulated in gut tissue from an E isolate-infected C57BL/6 mouse compared to tissue from an S isolate infection, including the message for the anti-microbial protein, angiogenin 4 (Ang4). This led to the identification of distinct differences in the goblet cell phenotype post-infection with the two isolates.ConclusionDifferences in gene expression levels identified between the S and E-infected mice early during infection have furthered our knowledge of how the S isolate persists for longer than the E isolate in the C57BL/6 mouse. Potential new targets for manipulation in order to aid expulsion have been identified. Further we provide evidence for a potential new marker involving the acidity of the mucins within the goblet cell which may predict outcome of infection within days of parasite exposure.

Isolates of Trichuris muris elicit different adaptive immune responses in their murine host

The J and S isolates of Trichuris muris have different infection profiles in C57BL/6 mice; J worms are expelled, S worms survive to chronicity. Building on this, the ability of the J and S isolates to survive, and the quality of the immune response induced was explored in three different strains of mouse. The resistant BALB/c mouse mounted a strong Th2 response against both isolates, which were quickly expelled. The susceptible AKR host mounted a Th1 response and retained both isolates. Despite equivalent worm exposure, mesenteric lymph node cells from AKR mice infected with the S isolate produced significantly higher levels of IL-12 and the intestinal mastocytosis was reduced. IgG1 and IgG2a from S-infected AKR mice recognized low molecular weight antigens not recognized by J-infected mice. Differential expulsion kinetics was observed in the slower-responding C57BL/6 strain; J worms were expelled but S isolate worms were retained. Survival of the S isolate was again associated with elevated IL-12 and decreased Th2 responses. In resistant mouse strains, the outcome of infection is thus dominantly influenced by host genetics. However, in the slower-responding host, isolate-derived factors may play a role in shaping the quality of the adaptive immune response, thus influencing parasite survival.

NK1.1+ cell depletion in vivo fails to prevent protection against infection with the murine nematode parasite Trichuris muris.

Protection against the murine nematode parasite Trichuris muris has been shown to involve interleukin 4 (IL-4). NK1.1+ T cell receptor alphabeta+ cells, designated Natural Killer T (NKT) cells, produce a large amount of IL-4 in response to anti-CD3 stimulation and numerous pieces of evidence suggest that NKT cells provide the initial source of IL-4 for T helper 2 (Th2) priming. These observations allow the hypothesis that NKT cells produce a large amount of IL-4 in response to T. muris infection and augment Th2 responses and IL-4 production, thus achieving protection against T. muris. To investigate the involvement of NKT cells in protection against T. muris infection, NK1.1+ cell-depleted B10.BR mice were prepared by anti-NK1.1 monoclonal antibody injection. Efficient expulsion of T. muris worms occurred in NK1.1+ cell-depleted infected mice, and the expulsion kinetics of T. muris worms, the levels of IL-4 production by mesenteric lymph node cells, and the kinetics of the specific IgG1 and IgG2a responses to T. muris were similar to those in mouse IgG-treated or non-treated control B10.BR mice. These observations suggest that NK1.1+ cells and NKT cells are not involved in the induction of Th2 responses and protective immunity to T. muris infection.

Genetic difference in susceptibility and fatality of three strains of mice experimentally infected with Neodiplostomum seoulense.

The genetic influence on host susceptibility to Neodiplostomum seoulense infection and fatality of the host was studied in 3 inbred strains of mice (BALB/c [H-2d], C3H/He [H-2k], and C57BL/6 [H-2b]). The survival of the mice, worm expulsion kinetics, worm size, number of eggs produced per day (EPD), and number of uterine eggs were observed from day 1 to day 40 postinfection (PI) with 100 or 200 metacercariae per mouse. Infection with N. seoulense was highly lethal to all 3 strains of mice, but the lethality was dose-dependent and varied according to the genetic backgrounds of the mice. The C3H/He mice exhibited the highest mortality, the lowest worm burdens and EPD, and the quickest expulsion of worms. It is suggested that different genetic backgrounds of mice appear to affect the host's capacity to expel N. seoulense and the fatality of the hosts themselves.

Avermectin\\milbemycin resistance in trichostrongyloid nematodes

Resistance to levamisole and the benzimidazoles appears to be achieved by one or, at most, two mechanisms in the common trichostrongyloid parasites of sheep. For the avermectin\\milbemycin anthelmintic class the picture is more complex. In-vitro assays employing the free-living stages of trichostrongyloid nematodes were used to investigate structure–activity relationships for the avermectins\\milbemycins. While avermectin\\milbemycin-susceptible isolates of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta were found to differ in their intrinsic sensitivities to avermectin\\milbemycin inhibition of larval development and L3 motility, structure–activity profiles against all three species were similar. In-vivo avermectin\\milbemycin resistance was associated with a reduced sensitivity to avermectin\\milbemycin inhibition of larval motility and\\or development in some, but not all, isolates. Where a reduced sensitivity to avermectin\\milbemycin inhibition of larval development was observed, different groups of resistant isolates displayed different structure–activity profiles. Many avermectin\\milbemycin-resistant isolates showed an increased sensitivity to paraherquamide. These in-vitro data have allowed the classification of avermectin\\milbemycin-resistant isolates into a number of distinct types. Study of the inheritance of avermectin\\milbemycin resistance in two resistance types suggests that the in-vitro differences between resistant isolates reflect important differences in the mechanism of resistance present. The kinetics of expulsion of H.contortus, T. colubriformis and O. circumcincta from sheep following treatment with ivermectin indicate that, in vivo, the critical action of avermectins\\milbemycins against O.circumcincta may be different to that which results in H. contortus and T. colubriformis elimination. This observation provides some explanation for the differences between resistant isolates. If, for different species, the critical event(s) leading to expulsion are different, then it follows that the mechanisms of resistance observed may also differ.

Kinetics of expulsion of Haemonchus contortus from sheep and jirds after treatment with closantel

Experiments were conducted in sheep after intramuscular treatment with closantel and in jirds after oral treatment with closantel to determine when expulsion of established H.contortus commences. Expulsion starts at about 8 h in sheep and coincides with the onset of reduced motility in worms recovered from the abomasum. In jirds, expulsion starts by 2 h after treatment. Experiments also conducted in jirds showed that infective larvae are first killed by circulating closantel 3 days after infection, when blood feeding starts, and that by 8 days 80% of larvae are lost.

Characterization of Nippostrongylus brasiliensis Infection in Different Strains of Mice

Five mouse strains, CBA/J, BALB/c, C3H/HeJ, A/J, and C57Bl/6J-bg-bg, all showed similar expulsion kinetics for Nippostrongylus brasiliensis (infective dose = 500 L3). Typically, parasite recovery was maximal on day 2 in the lungs and by day 4 in the small intestine. Few worms (less than 5% infective dose) were recovered on day 14 in all strains. These same mouse strains exhibited immune depression on day 5 of infection with mesenteric lymph node cells (MLN) showing reduced (10-30% normal) IgM, IgG, and IgA responses against heterologous antigen. The intestinal mast cell numbers and tissue histamine levels were examined in CBA/J mice. Mast cell numbers increased (normal = less than 1/villous crypt unit; VCU) from day 5 and peaked on day 12 (greater than 15/VCU). Intestinal histamine levels did not completely correlate with mast cell numbers with maximum concentrations (240 +/- 73 ng/g, 2-fold over normal) reached by day 8. Histamine concentrations in the intestine returned to normal levels by day 20.

Characteristics of Hydrocarbon Deposits in the Green Tuff

Several hydrocarbon deposits have been found in volcanic rocks of the Nanatani stage (so-called Green Tuff) in the Niigata basin, Japan.Their characteristics are summarized as follows.1) Hydrocarbon deposits are formed in anticlinal structures and are mostly located close to depressions.2) Oil accumulations have been found in anticlinal structures which are formed in the early stage.3) All hydrocarbon deposits except for the Shiunji gas field are located in the Chuetsu sub-basin.4) Larger oil and gas fields are located in or close to the kitchen areas of oil and gas. This suggests the short lateral migration of hydrocarbons together with the distribution of oil and gas in neighboring areas of the Mitsuke oil field.5) In larger gas fields, the Teradomari formation shows a typical abnormally high pressur e, whereas the Geen Tuff is hydrostatical or slightly highly-pressured. On the other hand, formation pressure in the Mitsuke oil field increases toward lower straigraphic horizons that is similar to the pressure distribution in the Shiiya and Nishiyama reservoirs. These pressure patters are closely related to the manner of accumulation of oil and gas.6) The migration of oil and gas in larger oil and gas fields is thought to have occurred during and after the Nishiyama stage.7) The distributions of oil and gas in the Green Tuff depend rather on the kinetics of hydrocarbon expulsion from source rocks than on the type of organic matter. The kinetics of expulsion is controlled by the relative positions of the hydrocarbon generation zone and the overpressured zone.The Niigata basin can be sub-divided into three parts according to their geologic conditions: Kaetsu sub-basin, Kakuta-Sanjo uplift and Chuetsu sub-basin. Considering the combination of trap, source rock, reservoir and migration mechanism, it is concluded that the Chuetsu sub-basin is the most prospective for future exploration.

Kinetics of expulsion of the nematode, Nippostrongylus brasiliensis, in mast-cell deficient W/WV mice.

Mucosal mast-cell hyperplasia is frequently observed in intestinal nematode infections and it has been suggested that mast-cell responses to parasite antigens are involved in worm expulsion (self cure). To evaluate the importance of this mechanism, the course of infection and expulsion of Nippostrongylus brasiliensis was compared in mast-cell deficient W/WV and normal (+/+) mice. Initial infectivity rates were similar, but the subsequent kinetics of expulsion of adult worms differed principally in that the onset of expulsion in mast-cell deficient mice appeared to occur 24-36 h later than that in normal mice. Expulsion was complete by the 14th day post infection in both W/WV and normal mice. Worm fertility (as estimated by faecal egg output) also differed in W/WV and normal mice, with maximal egg output in W/WV mice occurring 24 h later than that in normal mice. Although a few mast cells were present in the intestinal mucosa and tongue of W/WV mice, their numbers did not change during the course of infection with N. brasiliensis. In contrast, worm expulsion in normal mice was associated with a moderate increase in numbers of intestinal mast cells, commencing at the onset of expulsion and peaking several days after expulsion was completed.

Comparison of the Kinetics of Expulsion of Trichostrongylus colubriformis from Previously Uninfected, Reinfected, and Vaccinated Guinea Pigs

Comparison of the Kinetics of Expulsion of Trichostrongylus colubriformis from Previously Uninfected, Reinfected, and Vaccinated Guinea Pigs

Studies on the kinetics of expulsion by guinea-pigs of primary infections with the intestinal nematode parasite Trichostrongylus colubriformis

Heston strain guinea-pigs were infected with 1000 Trichostrongylus colubriformis infective larvae and the kinetics of parasite expulsion from the host studied. Expulsion began approximately 15 days after infection, after which log worm count declined from an initial level of approximately 2·77 at a rate of approximately 0·1 per day. There was no significant difference in worm counts in each sex.