Quantitative trait loci for resistance to Heligmosomoides bakeri and associated immunological and pathological traits in mice: comparison of loci on chromosomes 5, 8 and 11 in F2 and F6/7 inter-cross lines of mice.

A comparison of F2 and F6/7 inter-cross lines of mice, derived from CBA and SWR parental strains, has provided strong evidence for several previously undetected quantitative trait loci (QTL) for resistance to Heligmosomoides bakeri. Five QTL affecting average faecal egg counts and/or worm burdens in week 6 were detected on mouse chromosomes 5 (Hbnr9 and Hbnr10), 8 (Hbnr11) and 11 (Hbnr13 and Hbnr14). Three QTL for faecal egg counts in weeks 4 and 6 were found on both chromosomes 5 (Hbnr9) and 11 (Hbnr13 and Hbnr14). Two QTL for the mucosal mast cell protease 1 (MCPT1) response were located on chromosomes 8 (Hbnr11) and 11 (Hbnr13), two for the IgG1 antibody response to adult worms on chromosomes 5 (Hbnr10) and 8 (Hbnr11), two for PCV in week 6 on chromosomes 5 (Hbnr9) and 11 (Hbnr13), and two for the granulomatous response on chromosome 8 (Hbnr12) and 11 (Hbnr15). Our data emphasize that the control of resistance to H. bakeri is multigenic, and regulated by genes within QTL regions that have a complex range of hierarchical relationships.

Prolonged elevation of galanin and tachykinin expression in mucosal and myenteric enteric nerves in trinitrobenzene sulphonic acid colitis.

Diverticulitis causes recurrent abdominal pain associated with increased mucosal expression of mucosal galanin and substance P (SP). We studied changes in mucosal and myenteric plexus neuropeptides in adult rats using a model of colonic inflammation, trinitrobenzenesulphonic acid colitis. We assessed the effects on the pan-neuronal markers protein gene product 9.5 (PGP9.5) and neurofilament protein, as well as specific neuropeptides at 1, 2, 3, 4, 6, 8, 10 and 14 weeks. Following the acute injury there was macroscopic resolution of inflammation but minor microscopic abnormalities persisted. Percent area stained of mucosal PGP9.5 fell initially but average levels on days 21 and 28 levels were significantly elevated (P < 0.001), returning to normal by day 42. Percent area staining of PGP9.5 in the muscle rose immediately and remained significantly elevated at 70 days (P < 0.001). SP, neuropeptide K and galanin followed a similar overall pattern. SP to PGP9.5 ratio was significantly increased in the muscle both acutely (days 1-28) and in the long term (days 70 and 98), whereas the galanin to PGP9.5 ratio was significantly increased in the mucosa throughout the study. Low-grade chronic inflammation after an acute initial insult causes a persistent increase in the expression of galanin in the mucosa and SP in muscle layer.

Naturally occurring variability in some phenotypic markers and correlates of haemonchotolerance in West African Dwarf goats in a subhumid zone of Nigeria.

West African Dwarf (WAD) goats of the Nigerian subhumid zone generally show strong resistance and resilience to Haemonchus contortus in laboratory experiments, although a relatively small proportion are susceptible to infection. Little is known about these extremes of response phenotype in nature. Therefore, a survey was carried out of gastrointestinal nematode infections in WAD goats, with emphasis on abomasal worms, at three goat markets in Southern Nigeria during the rainy season. Faecal samples (n=1070) were collected weekly from goats between April and September, and 352 abomasa and small intestines from local abattoirs were examined. Total strongyle (prevalence=65.0%) and H. contortus (prevalence=64.3%) faecal egg counts (FEC) varied between the three markets, being highest throughout at Opi. FEC increased from April to peak in August. Based on raw FEC, 76.1% of goats had FEC of <100, and 4.7%>500. Adjustment of these figures for monthly and between-market differences, gave figures of 78.8 and 3.4%, respectively. H. contortus worm burdens (WB) showed a similar pattern with 67.9% of goats harbouring <200 worms and 8.2% >1000, and after adjustment 69.6 and 6.0%, respectively. Fecundity, based on eggs in the uterus, did not vary between markets or monthly, but fell with increasing WB. Trichostrongylus colubriformis was less frequent (prevalence=42.4%) but goats from Opi also carried higher WB, and worms were similarly highly aggregated in hosts. When the between-market and monthly differences for both species were controlled, a highly significant positive correlation between the species emerged. Therefore, although a small subset of goats, highly susceptible to H. contortus, exists in this breed, the majority show resistance under field conditions and the resistant phenotype is also resistant to T. colubriformis. Both species are highly aggregated in the susceptible subset of the population. While, we cannot yet exclude alternative explanations, our data are compatible with a strong genetic basis for this phenomenon.

Genetic variation in resistance to repeated infections with Heligmosomoides polygyrus bakeri, in inbred mouse strains selected for the mouse genome project.

Since the publication of the mouse genome, attention has focused on the strains that were selected for sequencing. In this paper we report the results of experiments that characterized the response to infection with the murine gastrointestinal nematode Heligmosomoides polygyrus of eight new strains (A/J, C57BL/6, C3H, DBA/2, BALB/c, NIH, SJL and 129/J), in addition to the well-characterized CBA (poor responder) and SWR (strong responder) as our controls. We employed the repeated infection protocol (consisting of 7 superimposed doses of 125L3 each administered at weekly intervals, faecal egg counts in weeks 2, 4 and 6 and assessment of worm burdens in week 6) that was used successfully to identify quantitative trait loci for genes involved in resistance to H. polygyrus. SWR, SJL and NIH mice performed indistinguishably and are confirmed as strong responder strains to H. polygyrus. CBA, C3H and A/J mice all tolerated heavy infections and are assessed as poor responders. In contrast, DBA/2, 129/J and BALB/c mice performed variably between experiments, some tolerating heavy worm burdens comparable to those in poor responders, and some showing evidence of resistance, although only in one experiment with female 129/J females and one with female BALB/c was the pattern and extent of worm loss much like that in SWR mice. Because the genetic relationships between six of the strains exploited in this study are now well-understood, our results should enable analysis through single nucleotide polymorphisms and thereby provide more insight into the role of the genes that control resistance to H. polygyrus.

Enterochromaffin cell hyperplasia and decreased serotonin transporter in a mouse model of postinfectious bowel dysfunction.

Patients with postinfective irritable bowel syndrome and Trichinella spiralis-infected mice share many features including visceral hypersensitivity and disordered motility. We assessed enterochromaffin (EC) numbers and serotonin transporter (SERT) using National Institute of Health (NIH) female mice studied for up to 56 days post-T. spiralis infection. The effects of steroid treatment and the T-cell dependence of the observed responses were assessed by infection of hydrocortisone-treated or T-cell receptor knock out [TCR (betaxdelta) KO] animals. Enterochromaffin cell density in uninfected animals increased from duodenum 10.0 cells mm-2 (5.9-41.0) to colon 61.8. (46.3-162) cells mm-2 P<0.0001. Infection increased duodenal and jejunal counts which rose to 37.3 (22-57.7) cells mm-2 and 50.6 (7-110.8) cells mm-2, respectively, at day 14. Infection significantly reduced jejunal SERT expression, with luminance values falling from 61.0 (45.1-98.3) to a nadir of 11.6 (0-36.0) units at day 9, P<0.001. Specific deficiencies in all T cells reduced EC hyperplasia and abrogated infection-induced mastocytosis. Thus infection induced inflammation increases EC numbers, as has been reported in PI-IBS, and reduces SERT. This may increase mucosal 5HT availability and contribute to the clinical presentation of PI-IBS.

The modulatory influence of Trypanosoma brucei on challenge infection with Haemonchus contortus in Nigerian West African Dwarf goats segregated into weak and strong responders to the nematode.

Although Nigerian West African Dwarf (WAD) goats are relatively resistant to infection with Haemonchus contortus and are also trypanotolerant, natural outbreaks of both infections are known to occur. Despite their relative resistance to H. contortus WAD goats nevertheless show variability in response phenotype and it was of interest to examine the effect of this variability on the outcome of concurrent trypanosome infection. Trypanosoma brucei infections were established in goats that were initially classified as good or poor responders to H. contortus. Thirty-nine goats were exposed to an escalating infection with H. contortus, and on the basis of their mean faecal egg counts (FEC) were allocated to high FEC (poor responders, 18 goats with the highest FEC) or low FEC (good responders, 18 goats with the lowest FEC) classes. Nine uninfected naive control goats were included to provide reference baseline values. Retrospective analysis of parasitological and pathological parameters after allocation into high/low FEC classes showed that FECs differed significantly, in both classes packed cell volume (PCV) values fell relative to naive controls, neither class lost weight and both generated marked IgG responses. All goats received anthelmintic on day 61, half of each group was infected with 50 million trypanosomes and on day 67, excepting the controls, all goats were challenged with 3000 L3 of H. contortus. Trypanosome parasitaemia was generally low, and marginally, but not significantly, higher in the low compared with high FEC class, peaking 12-16 days after exposure in both groups and then falling to below microscopically detectable levels (although still detectable by sub-inoculation into mice) by week 3. At autopsy (days 109/110), worm burdens were significantly higher in the trypanosome-infected goats from the high FEC class, relative to all other groups. Trypanosome infected goats showed a tendency (although not significant) towards higher FEC and, irrespective of their FEC class, had lower PCV values although body weight did not vary significantly. All goats challenged with H. contortus had higher antibody levels than naive controls, but neither trypanosome infection nor FEC class affected the magnitude of responses. These results confirm that WAD goats comprise a range of response phenotypes to initial H. contortus infection and that trypanotolerance is a key trait of this breed. Although immunity to nematode infection develops even in poor responders, these animals harbour higher nematode burdens during concurrent infection with T. brucei.

Variability in the resistance of the Nigerian West African Dwarf goat to abbreviated escalating trickle and challenge infections with Haemonchus contortus.

The West African Dwarf (WAD) goat is known to be relatively resistant to infection with Haemonchus contortus. An experiment was carried out to assess variability in resistance under escalating trickle and single pulse challenge infection protocols. Thirty two 7-8 month old goats were subjected to trickle infection beginning with a thrice weekly dose of 100 L3 and increasing to 500 L3 by week 5, the last dose being administered on D30. Infection was abbreviated with fenbendazole on D32 and the animals challenged with 4000 L3 (=404-741L3/kg body weight) on D46 and necropsied on D81. The goats exhibited marked individual variability in faecal egg counts (FEC) and worm burdens (Wb). For convenience, we arranged the goats into FEC classes 1, 2 and 3, corresponding to peak counts of <1000, 1000-2500 and >2500 epg and Wb classes 1, 2 and 3, with counts of <50, 50-100 and >100 worms, respectively, broadly reflecting relatively resistant, intermediate and poorly resistant response phenotypes. Wb classes broadly reflected the FEC phenotypes and although there were no significant differences between the Wb of the FEC classes at necropsy, significant differences in FEC were detected when analysis was carried out only on Wb classes. There was also a significant positive correlation between FEC class on each of the days of challenge (D67 to D80) and worm burden at necropsy. In general, the goats showed strong resistance/tolerance to challenge, with more than 80% having Wb not exceeding 5% of the challenge dose, and only in class three goats were weight loss and anaemia detected in some animals. PCV and weight changes also closely reflected the FEC and Wb classes, with a significant negative correlation between each and Wb. Infection level did not significantly affect the sizes of either male or female worms and no significant difference was detected between the levels of circulating eosinophils in any FEC or Wb class. Overall, the data are consistent with the existence of a range of response phenotypes, which reflect variations in resistance of WAD goats to H. contortus infection, the genetic basis of which will be explored in future experiments.

Effect of priming/booster immunisation protocols on immune response to canine parvovirus peptide induced by vaccination with a chimaeric plant virus construct.

Expression of a 17-mer peptide sequence from canine parvovirus expressed on cowpea mosaic virus (CPMV) to form chimaeric virus particles (CVPs) creates vaccine antigens that elicit strong anti-peptide immune responses in mice. Systemic (subcutaneous, s.c.) immunisation and boosting with such CVP constructs produces IgG(2a) serum antibody responses, while mucosal (intranasal, i.n.) immunisation and boosting elicits intestinal IgA responses. Combinations of systemic and mucosal routes for priming and boosting immunisations were used to examine their influence on the level, type and location of immune response generated to one of these constructs (CVP-1). In all cases, s.c. administration, whether for immunisation or boosting, generated a Th1-biased response, reflected in a predominantly IgG(2a) serum antibody isotype and secretion of IFN-gamma from in vitro-stimulated lymphocytes. Serum antibody responses were greatest in animals primed and boosted subcutaneously, and least in mucosally vaccinated mice. The i.n. exposure also led to IFN-gamma release from in vitro-stimulated cells, but serum IgG(2a) was significantly elevated only in mice primed intranasally and boosted subcutaneously. Peptide- and wild-type CPMV-specific IgA responses in gut lavage fluid were greatest in animals exposed mucosally and least in those primed and boosted subcutaneously or primed subcutaneously and boosted orally. Lymphocytes from immunised mice proliferated in response to in vitro stimulation with CPMV but not with peptide. The predominant secretion of IFN-gamma from all immunising/boosting combinations indicates that the route of vaccination and challenge does not alter the Th1 bias of the response to CVP constructs. However, optimal serum and intestinal antibody responses were achieved by combining s.c. and i.n. administration.

Chasing the genes that control resistance to gastrointestinal nematodes.

The host-protective immune response to infection with gastrointestinal (GI) nematodes involves a range of interacting processes that begin with recognition of the parasite's antigens and culminate in an inflammatory reaction in the intestinal mucosa. Precisely which immune effectors are responsible for the loss of specific worms is still not known although many candidate effectors have been proposed. However, it is now clear that many different genes regulate the response and that differences between hosts (fast or strong versus slow or weak responses) can be explained by allelic variation in crucial genes associated with the gene cascade that accompanies the immune response and/or genes encoding constitutively expressed receptor/signalling molecules. Major histocompatibility complex (MHC) genes have been recognized for some time as decisive in controlling immunity, and evidence that non-MHC genes are equally, if not more important in this respect has also been available for two decades. Nevertheless, whilst the former have been mapped in mice, only two candidate loci have been proposed for non-MHC genes and relatively little is known about their roles. Now, with the availability of microsatellite markers, it is possible to exploit linkage mapping techniques to identify quantitative trait loci (QTL) responsible for resistance to GI nematodes. Four QTL for resistance to Heligmosomoides polygyrus, and additional QTL affecting faecal egg production by the worms and the accompanying immune responses, have been identified. Fine mapping and eventually the identification of the genes (and their alleles) underlying QTL for resistance/susceptibility will permit informed searches for homologues in domestic animals, and human beings, through comparative genomic maps. This information in turn will facilitate targeted breeding to improve resistance in domestic animals and, in human beings, focused application of treatment and control strategies for GI nematodes.

Upregulation of the oncogene c-myc in Barrett's adenocarcinoma: induction of c-myc by acidified bile acid in vitro.

BACKGROUND AND AIMS: C-myc over expression is implicated in malignancy although to date this has not been studied in Barrett's metaplasia. We sought to determine c-myc expression in the malignant progression of Barrett's metaplasia and whether it may be induced by bile acids seen in gastro-oesophageal refluxate. METHODS: C-myc protein and mRNA levels were assessed in 20 Barrett's metaplasia and 20 oesophageal adenocarcinoma samples by western blotting and real time polymerase chain reaction. Levels of c-myc and proliferation were also assessed in cell lines OE21, OE33, SW-480, and TE-7 stimulated with pulses or continuous exposure to the bile acids deoxycholic acid and chenodeoxycholic acid. RESULTS: C-myc protein was upregulated in 50% of Barrett's metaplasia and 90% of oesophageal adenocarcinoma samples compared with squamous, gastric, and duodenal controls. C-myc immunolocalisation in Barrett's metaplasia revealed discrete nuclear localisation, becoming more diffuse with progression from low to high grade dysplasia to adenocarcinoma. Both continual and pulsed bile acid induced c-myc at pH 4, with no effect at pH 7 or with acidified media alone. Pulsed bile acid treatment induced proliferation (p<0.05); in contrast, continuous exposure led to suppression of proliferation (p<0.05). CONCLUSIONS: We have shown upregulation of c-myc with malignant progression of Barrett's metaplasia and suggest that acidified bile may be a novel agent responsible for induction of this oncogene.

Trypanosome-induced modulation of responses to concurrent helminth infection.

Infections with African trypanosomes are known to suppress immune responses to vaccines and to gastrointestinal nematode infections in livestock. Experimental infections with Trypanosoma brucei (Tb) and the gastrointestinal nematode Nippostrongylus brasiliensis (Nb) in mice were used to identify possible mechanisms involved in interference with anti-worm responses and to examine the effects of host genotype on the extent of suppression seen. Concurrent infections with T. brucei resulted in a prolongation of worm survival and a dramatic increase in faecal egg output. Infection also resulted in a marked suppression of the proliferative response of mesenteric lymphocytes (MLNC) to in vitro mitogenic stimulation. When MLNC from concurrently infected mice were stimulated in vitro with the mitogen ConA they released more IFN-gamma and less IL-5 than cells from mice infected only with N. brasiliensis. These data are interpreted in terms of a trypanosome-mediated influence on the development of host-protective type-2 T helper cell responses against N. brasiliensis. The degree to which T. brucei altered the kinetics of the nematode infection was influenced by the particular mouse strain concerned.

Paneth and intermediate cell hyperplasia induced in mice by helminth infections.

Hyperplasia of Paneth and intermediate cells is a recently described component of the response of the small intestine of mice to infection with the nematode Trichinella spiralis. To investigate whether this hyperplasia is parasite specific or represents a generic intestinal response to infection, mice were infected with T. spiralis, Nippostrongylus brasiliensis, Heligmosomoides polygyrus or Schistosoma mansoni and tissue samples taken at various time-points post-infection to determine Paneth and intermediate cell numbers. All infections induced Paneth and intermediate cell hyperplasia, but the patterns of response varied between the parasite species concerned, reflecting differences in their relationships with the host. Increases in the numbers of these cells appeared to correlate with known patterns of T-helper-2 immune responses.

Characterization of the immune response to canine parvovirus induced by vaccination with chimaeric plant viruses.

NIH mice were vaccinated subcutaneously or intranasally with chimaeric cow pea mosaic virus (CPMV) constructs expressing a 17-mer peptide sequence from canine parvovirus (CPV) as monomers or dimers on the small or large protein surface subunits. Responses to the chimaeric virus particles (CVPs) were compared with those of mice immunized with the native virus or with parvovirus peptide conjugated to keyhole limpet haemocyanin (KLH). The characteristics of the immune response to vaccination were examined by measuring serum and mucosal antibody responses in ELISA, in vitro antigen-induced spleen cell proliferation and cytokine responses. Mice made strong antibody responses to the native plant virus and peptide-specific responses to two of the four CVP constructs tested which were approximately 10-fold lower than responses to native plant virus. The immune response generated by the CVP constructs showed a marked TH1 bias, as determined by a predominantly IgG(2a) isotype peptide-specific antibody response and the release of IFN-gamma but not IL-4 or IL-5 from lymphocytes exposed to antigen in vitro. In comparison, parvovirus peptide conjugated to KLH generated an IgG(1)-biased (TH2) response. These data indicate that the presentation of peptides on viral particles could be used to bias the immune response in favor of a TH1 response.Anti-viral and anti-peptide IgA were detected in intestinal and bronchial lavage fluid of immunized mice, demonstrating that a mucosal immune response to CPV can be generated by systemic and mucosal immunization with CVP vaccines. Serum antibody from both subcutaneously-vaccinated and intranasally-vaccinated mice showed neutralizing activity against CPV in vitro.

Parameters of intestinal inflammation in mice given graded infections of the nematode Trichinella spiralis.

Four parameters of the intestinal inflammatory response (numbers of mucosal mast cells (MMC) and Paneth cells, villus:crypt ratios and mitotic figures) were measured in mice exposed to varying doses of infective larvae of Trichinella spiralis. The aim of the experiments was to determine whether generation of these components of inflammation required a threshold level of infection and whether, once triggered, inflammation became pan-mucosal. Near maximal MMC and Paneth cell responses were elicited even with infections as low as 35 larvae; changes in villus:crypt ratios and in mitotic indices also occurred at this level of infection, but were progressively greater with increasing levels of infection. In all infected mice, including those infected with 35 larvae, MMC and Paneth cell responses extended over most of the small intestine. These data are interpreted as showing: (i) that the intestinal mucosa is highly responsive to T. spiralis infection; (ii) that once triggered, components of the inflammatory response are amplified by T cell-dependent mechanisms, becoming pan-mucosal; and (iii) that MMC and Paneth cell responses, which require cell division and differentiation, become maximal at a lower infection threshold than changes in the villus:crypt ratio or in mitotic indices, which directly reflect increased rates of division in crypt cells.

Tyvelose and protective responses to the intestinal stages of Trichinella spiralis.

The unusual sugar tyvelose is the immunodominant portion of the major larval glycoprotein antigens of Trichinella spiralis, which play an important role in generating immunity against the intestinal stages of infection. The possibility that the tyvelose component itself may have a host- or parasite-protective role in the intestine was tested by following the outcome of challenge infections in mice primed and boosted with tyvelose-BSA, or in mice primed with tyvelose-BSA before boosting with larval antigen. Although antibody responses were raised against tyvelose there was no evidence of protective immunity against the intestinal stages, as assessed by total adult worm recovery or by size and fecundity of female worms in immunized mice. Equally, priming with tyvelose-BSA before boosting with larval antigen had no effect on the expression of immunity against a challenge infection. The predominant antibody isotype recorded in all immunized mice was IgG1, suggesting the induction of type 2 T cell responses, and this was confirmed by cytokine analysis, mesenteric node lymphocytes of all mice showing production of IL-5 but not IFN-gamma. Clearly immunization with tyvelose had no significant effect on T cell polarization. The data show that, with the experimental design employed, there was no evidence for a functional role of tyvelose in either host- or parasite-protection during the intestinal phase of infection.

Expression of acquired immunity to a local isolate of Haemonchus contortus by the Nigerian West African Dwarf goat.

The capacity of young Nigerian West African Dwarf (WAD) goats to express good acquired immunity to their native geographic strain of Haemonchus contortus and the correlates of this responsiveness were studied in a laboratory experiment involving forty 7-8 month old kids. A primary immunising infection with 2000 L3 (equivalent to 260-450 L3/kg body weight) with or without challenge on D42 with 2000 L3 resulted in a mild chronic infection with a pre-patent period of 18-20 days and little or no reduction in worm burden between D14 and D56. In contrast, another group (D) of kids, whose immunising infection had been truncated with fenbendazole on D35 and later received similar challenge infection, developed good protection against challenge. Thus, worm burdens were largest in group E (challenge control), larger in group C (primary+challenge) and least in group D. Of the measures of infection used, namely faecal worm egg counts (FECs), circulating eosinophil (EOS) responses, packed cell volume (PCV) and body weight, FEC and EOS responses exhibited marked individual variability, but only FEC (geometric mean of transformed counts) and PCV showed strong correlation with worm burden. There was also a significant negative correlation between FEC and PCV. The size of inoculum used was well tolerated by the kids, as it induced only mild changes in PCV in some goats and no effect at all on body weights. This suggests that the WAD goat may possess a good measure of resistance to the pathogenic effects of its native strain of H. contortus. The wide individual variability in FEC and its strong relationships to worm burden and PCV are pointers to its likely genetic basis. There are, therefore, good prospects for further studies to identify H. contortus resistant genotypes among the WAD goat population.

Variation and immunity to intestinal worms.

Genetically determined variation in host capacity to express resistance to a given parasite plays a major role in determining the outcome of infection. It can be assumed that the same is true of variation in parasites, but very much less is known of its influence on the host-parasite relationship. Phenotypic and genotypic variation within species of intestinal worms is now well documented, detailed studies having been made of parasites such as Ascaris in humans and trichostrongyles in domestic animals. However, the extent to which this variation affects the course of infection or the host immune response in these hosts is limited. Of the nematodes used as experimental models in laboratory rodents, detailed data on phenotypic or genotypic variation are limited to Strongyloides and Trichinella. Parasite variation is known to be subject to host-mediated selection, the emergence of anthelmintic resistance being a good example. Repeated passage has been used to select lines of parasite that survive in abnormal hosts or which show adaptation to host immunity. Experimental studies with Trichinella genotypes in mice have demonstrated the extent to which parasite variation influences the nature and degree of the host's immune and inflammatory responses, the complex interplay between immunogenicity and pathogenicity influencing both partners in the relationship. Recent studies with isolates of Trichuris muris have shown how parasite variation influences the capacity of mice to express the T helper cell responses necessary for resistance. Molecular differences between T. muris isolates have been shown in their excreted/secreted products as well as at the level of their DNA. Knowledge of the functional consequences of parasite variation will add to our understanding of host-parasite evolution as well as providing a rational basis for predicting the outcome of controls strategies that rest on the improvement of host resistance through vaccination or selective breeding.

Methods for the stable isotopic analysis of chlorine in chlorate and perchlorate compounds.

Chlorate and perchlorate compounds, used as herbicides, solid fuel propellants, and explosives, are increasingly recognized as pollutants in groundwater. Stable isotope characterization would permit both environmental monitoring of extent of remediation and forensic characterization. Stoichiometric reduction to chloride (greater than 98% yield), by Fe(II) for chlorate and alkaline fusion-decomposition for perchlorate, allows analysis by standard methods to give highly reproducible and accurate delta37Cl results (0.05/1000, 2 x standard error). Analysis of various compounds from different suppliers yielded delta37Cl values for chlorate samples near to +0.2/1000 (SMOC), but one has within-sample heterogeneity of 0.5/1000, possibly due to crystallization processes during manufacture. Results for perchlorate samples also are generally near +0.2/1000, but one is +2.3/1000 (SMOC). The initial results suggest that both forensic and environmental applications might be feasible.

Mucosal T cells regulate Paneth and intermediate cell numbers in the small intestine of T. spiralis-infected mice.

Secretions of Paneth, intermediate and goblet cells have been implicated in innate intestinal host defense. We have investigated the role of T cells in effecting alterations in small intestinal epithelial cell populations induced by infection with the nematode Trichinella spiralis. Small intestinal tissue sections from euthymic and athymic (nude) mice, and mice with combined deficiency in T-cell receptor beta and delta genes [TCR(beta/delta)-/-] infected orally with T. spiralis larvae, were examined by electron microscopy and after histochemical and lineage-specific immunohistochemical staining. Compared with uninfected controls, Paneth and intermediate cell numbers increased significantly in infected euthymic and nude mice but not infected TCR(beta/delta)-/- mice. Transfer of mesenteric lymph node cells before infection led to an increase in Paneth and intermediate cells in TCR(beta/delta)-/- mice. In infected euthymic mice, Paneth cells and intermediate cells expressed cryptdins (alpha-defensins) but not intestinal trefoil factor (ITF), and goblet cells expressed ITF but not cryptdins. In conclusion, a unique, likely thymic-independent population of mucosal T cells modulates innate small intestinal host defense in mice by increasing the number of Paneth and intermediate cells in response to T. spiralis infection.

Mucosal responses to infection with Trichinella spiralis in mice.

Infections with T. spiralis in mice elicit strong inflammatory responses. The nature and control of these responses, and their relationship to the process of worm expulsion, have been debated for many years. Many components of inflammation are, like worm expulsion, T cell-dependent, but some are not. The paper describes novel observations on Paneth cell responses to infection in immunologically normal mice and in a variety of T cell-deficient mice. Responses occurred normally in nu/nu and scid/scid mice but not in beta/delta knock out mice incapable of generating cells with functional TCR. However all of these mice failed to expel worms in the pattern seen in immunologically normal controls. These data are incorporated into a discussion of the causal relevance of intestinal inflammatory changes to the process of worm expulsion.