Trichinella Spiralis Infection Research Articles

Structural relationships between immune cells and longitudinal muscle during a Trichinella spiralis infection in the rat intestine.

A Trichinella spiralis infection produces an acute inflammatory reaction and tissue damage in the mucosa, and, in addition, functional changes occur in the external muscle layers. The aim of the present study was to characterize structural changes in the musculature that occur during early infection, and to identify relationships between immune cells and muscle cells, as part of an ongoing investigation into the immune modulation of motor function in the gut. Rats were infected with T. spiralis larvae and the gut fixed at 12 h, 24 h, 48 h and 6 days post-infection for electron microscopy of the longitudinal muscle. Macrophages and lymphocytes penetrated the longitudinal musculature 12-24 h post-infection. Distinct contacts were observed between specific cell types; cellular protrusions from macrophages or lymphocytes made close apposition contacts with smooth muscle cells. Resident macrophages in the subserosal space, fibroblast-like cells as well as smooth muscle cells showed marked activation during inflammation. Fibroblast-like cells were frequently seen intercalated between lymphocytes and smooth muscle cells, hence they may mediate communication between immune cells and the musculature.

Enhancement of resistance in mice treated with Lactobacillus casei: Effect on Trichinella spiralis infection

The ability of viable Lactobacillus casei, administered by an intraperitoneal route, to induce resistance in mice against Trichinella spiralis infection was tested. The percent reduction of adult worms in the intestine 5 days after T. spiralis infection observed in L. casei-treated animals, compared with those of the control group, fluctuated between 70.9 and 88.5%; reductions of larvae per gram of muscle tissue, evaluated at 30 days after infection, varied from 46.6 to 84.4% in L. casei-treated mice compared with controls. At 5 days after infection, intestinal villus height was shorter in T. spiralis-infected animals than in L. casei-treated or uninfected mice. There were fewer mast cells/crypt unit in L. casei-treated and uninfected mice than in T. spiralis-infected animals. Lactobacilli-treated mice showed twice as many mononuclear cells/crypt unit as T. spiralis-infected animals. The IFN- γ serum level was higher in L. casei-treated animals at the time of worm challenge as compared with T. spiralis-infected or uninfected mice. The results indicate that: (1) L. casei increased resistance that acted against T. spiralis infection in mice, (2) this resistance apparently was directed against adult worms in the intestine.

Diagnosis of porcine trichinellosis: parasitological and immunoserological tests in pigs from endemic areas of Argentina

In order to compare the reliability of serological and parasitological techniques for the diagnosis of porcine trichinellosis from endemic areas in Argentina, 116 pigs were studied: 61 animals from two separate outbreaks and 55 from a small abattoir. Direct diagnostic techniques included trichinoscopy and the artificial digestion method. Indirect diagnostic tests used in this study were the enzyme-linked immunosorbent assay (ELISA), employing the excretory–secretory products of muscle larvae (ML) as antigen, and the indirect immunofluorescence assay using as antigen ML in suspension (IIF-susp), cryostat sections of infected rat muscle or of free ML (IIF-slide). The percentage of parasitologically positive pigs was invariably lower than that of serologically positive animals (IIF-slide), even when digestion studies were carried out individually with a greater amount of muscle sample than required by current regulations. Close correlation was found between IIF using as antigen tissue sections and IIF using free ML sections, while IIF-susp proved unsuitable for diagnosis since this assay presented a high percentage of false negative results (20%). The IIF-slide technique proved positive in all parasitologically positive animals. ELISA rendered a lower percentage of positive reactions than IIF-slide, especially when worm burden was low. Since most parasitologically positive animals rendered at least two positive serological tests (two variations of IIF or IIF plus ELISA), those negative by digestion and positive by two serological methods were strongly suspected of having trichinellosis. Upon studying swine from a abattoir it was found that 9% of the pigs were positive when assayed by two serological techniques, but Trichinella spiralis infection could not be parasitologically confirmed. To sum up, serological methods may be used for screening all pigs and positive findings should be tested by the digestion method by analysing a greater quantity of pork than that required by current regulations, above all in areas with reported clinical trichinellosis in humans, to ensure that the pork is safe for human consumption.

Persistent intestinal neuromuscular dysfunction after acute nematode infection in mice

BACKGROUND & AIMS: Although most acute enteric infections in humans resolve, some herald the onset of chronic symptomatology and persistent gastrointestinal dysfunction--so-called postinfectious irritable bowel syndrome. This entity is poorly understood, and there are no animal models for testing hypotheses. The aim of this study was to investigate changes in intestinal neuromuscular function during and after recovery from acute intestinal inflammation due to primary Trichinella spiralis infections in NIH Swiss mice.METHODS: Morphometric scores and myeloperoxidase activity were used to monitor mucosal inflammation. Neuromuscular function was assessed in vitro by pharmacological or electrical stimulation of longitudinal muscle.RESULTS: Acute inflammation resulted in an approximately 50% reduction of villus height, an approximately 50% increase in crypt depth, and a threefold increase in myeloperoxidase activity. Carbachol- and KCl-induced contractions of longitudinal muscle were also increased threefold, whereas contraction induced by electrical field stimulation of intramural nerves was decreased by 60%. Mucosal morphology and myeloperoxidase activity rapidly returned to control values, but the increased muscle contractility and the decreased excitatory neurotransmission persisted as long as 42 and 28 days after infection, respectively.CONCLUSIONS: These findings show that transient mucosal inflammation alters enteric neuromuscular function; this alteration persists after recovery from the infection and mucosal restitution.(Gastroenterology 1997 Oct;113(4):1224-32)

Pattern of proinflammatory cytokine mRNA expression during Trichinella spiralis infection of the rat.

Trichinella spiralis occupies an intramulticellular niche in the small intestinal epithelium, and thus we examined the intestine and gut-associated tissues for proinflammatory cytokines during the infection. We document the patterns of interleukin-1 (IL-1), IL-6, gamma interferon, and tumor necrosis factor alpha mRNA expression in the duodenum, jejunum, Peyer's patches, mesenteric lymph node, spleen, and liver in T. spiralis-infected rats. By reverse transcription-PCR detection of mRNAs, IL-1beta was found increased in the jejunum but only on day 2. The jejunal IL-1beta increase was attributed to the epithelium by isolating epithelial cells and then depleting them of intraepithelial lymphocytes prior to analysis. The only cytokine for which mRNA was substantially increased in tissues later in infection was tumor necrosis factor alpha in the spleen and, to a lesser extent, in the mesenteric lymph node. In fact mRNA levels for some cytokines declined below uninfected levels in some organs during the infection. IL-1 may be important in the initiation of the intestinal inflammatory response to this infection.

Intestinal transport and catabolism of IgE: a major blood-independent pathway of IgE dissemination during a Trichinella spiralis infection of rats.

Previous work has shown that Trichinella spiralis-infected rats transport IgE from plasma to intestinal tissue and fluids. In this study we quantitate IgE transport to the gut and circulation during T. spiralis infection in rats. Total IgE levels in intestinal fluid from infected rats were elevated by 4 days post-infection (dpi), but were not elevated in serum and lymph until 7 dpi. IgE levels in intestinal fluid ranged from 1 to 6 microg between 10 and 21 dpi, and serum and lymph IgE levels ranged from 100 to 200 ng/ml. Immunoprecipitation of intestinal fluid and enterocyte lysate at 11 dpi showed a protein of 190 kDa that was recognized by mouse anti-rat IgE-MARE-1 in Western blots. This protein was removed from intestinal wash samples with anti-IgE (A2)-Sepharose. The half-life of intact IgE in the intestinal lumen of rats 10 days after infection was 3.25 min. In serum, the half-life of IgE was 5 h. Analysis of IgE production and consumption in 10-day T. spiralis infected rats showed that about 4.67 microg IgE/day entered the serum, while 2570.00 microg IgE/day entered the intestinal lumen. The IgE present in serum 10 days after T. spiralis infection originated in the gut and/or associated lymphoid tissue and was transported to the circulation via thoracic duct lymph. However, most IgE produced in the intestine was transported to the gut lumen at a rate that exceeded transport to plasma by a factor of several-hundredfold.

Effects of oral L-NAME during Trichinella spiralis infection in rats.

We investigated the involvement of nitric oxide in transmural jejunal alterations induced by Trichinella spiralis (T. spiralis) infection in rats. Rats were gavaged with either saline or T.spiralis larvae, and, 1 h later, rats were treated orally with water, NG-nitro-L-arginine methyl ester (L-NAME; 30 mg/kg), or NG-nitro-D-arginine methyl ester (D-NAME; 30 mg/kg) on a daily basis. Although not observed in jejunum from uninfected rats, inducible nitric oxide synthase (iNOS) mRNA was present in the mucosa and neuromuscular layers of jejunum from T. spiralis-infected rats. On day 6, T. spiralis-infected rats had a 6-fold decrease in transmural nitric oxide synthase activity, an 11-fold increase in plasma nitrite, and a 7-fold elevation in transmural myeloperoxidase (MPO) activity compared with uninfected controls. Intestinal smooth muscle cell hyperplasia and hypertrophy were only detected in the infected rats. L-NAME, but not D-NAME, treatment of infected rats for 6 days caused a pronounced increase in transmural iNOS mRNA expression, coinciding with significantly increased mucosal nitric oxide synthase activity. T. spiralis numbers in L-NAME-treated rats were significantly lower compared with the other two infected groups although L-NAME had no direct effect on T. spiralis viability in vitro. Furthermore, L-NAME treatment significantly reduced plasma nitrite and jejunal MPO but not intestinal smooth muscle cell hyperplasia or hypertrophy. In contrast, D-NAME treatment of infected rats significantly enhanced intestinal smooth muscle hyperplasia and hypertrophy. Taken together, these results suggest that alterations in the T. spiralis-infected jejunum are mediated, in part, by a suppression of nitric oxide synthase activity in the inflamed jejunum.

Interleukin 1β induces the expression of interleukin 6 in rat intestinal smooth muscle cells

Background/Aims: The increased expression of several cytokines, including interleukin 6 (IL-6), has recently been reported in a study of the longitudinal muscle and myenteric plexus layers of rat intestine following Trichinella spiralis infection. However, the putative cellular sources and the mechanism underlying the induction of IL-6 in these tissues are presently unknown. The aim of this study was to examine the ability of cultured smooth muscle cells from rat jejunum to produce IL-6 messenger RNA and protein and to investigate the underlying mechanism. Methods: Cultured smooth muscle cells were treated with human recombinant interleukin 1β (HrIL-1β). The level of IL-6 messenger RNA was estimated by polymerase chain reaction, and the released IL-6 protein was estimated by bioassay. Results: HrIL-1β induced IL-6 messenger RNA expression in the smooth muscle cells in a time- and concentration-dependent manner. This was accompanied by the secretion of IL-6 protein into the medium. The effect of HrIL-1β was blocked by the IL-1 receptor antagonist, by actinomycin D, or by prior boiling of the cytokine. Conclusions: These findings show that HrIL-1β interacts with its receptor on smooth muscle cells to induce transcription of the IL-6 gene and to cause the secretion of IL-6. These results indicate that intestinal smooth muscle cells are not only targets for but also a source of cytokine.

Synergism between interleukins 1β and 6 on noradrenergic nerves in rat myenteric plexus

Background/Aims: Because levels of interleukins 1β and 6 (IL-1β and IL-6) are elevated during intestinal Trichinella spiralis infection, they may mediate the changes in enteric neural function in that model. IL-1β suppresses norepinephrine release from the myenteric plexus, but the effect of IL-6 is unknown. Therefore, we investigated the effects of IL-6 alone and in combination with IL-1β on norepinephrine release. Methods: Longitudinal muscle myenteric plexus or myenteric nerve varicosity preparations from jejunum of noninfected rats were loaded with [3H]norepinephrine, and 3H release was measured after a preincubation with or without human recombinant IL-6, alone or in combination with human recombinant IL-1β. Results: 1 ng/mL of IL-6 augmented 3H release, 100 ng/mL suppressed 3H release, whereas 10 ng/mL had no effect. However, IL-6 (10 ng/mL) plus a subthreshold concentration of human recombinant IL-1β significantly suppressed 3H release, and this was abolished by adding anti-IL-6 antibody or an IL-1 receptor antagonist. Conclusions: Because 3H release reflects [3H]norepinephrine release, our results show that IL-6 exerts a dual effect on norepinephrine release. Furthermore, there is synergism between IL-1β and IL-6 resulting in suppression of norepinephrine release. Therefore, both cytokines may contribute to the suppression of norepinephrine release observed in the inflamed intestine.

Minimal immunological changes in structurally malformed rats after prenatal exposure to cyclophosphamide

In order to compare the sensitivities of morphological and immunological parameters in a teratology study, effects in day 20 rat fetuses were stuudied after a single exposure to the immunosuppressive cytostatic agent cyclophosphamide (CP) on either day 11 or day 15 of gestation. Teratological methods included evaluation of external and skeletal morphology. Furthermore histology, immunohistochemistry and flow cytometry were performed on fetal thymus, liver and spleen. Immune function was assayed using the Trichinella spiralis infection model. Treatment resulted in dose-dependent gross morphological malformations, and in addition in overt skeletal anomalies such as brachygnathia, wavy ribs, and lordosis. In contrast, the immunological parameters tested revealed only minimal differences between treated and control groups. These results suggest either a remarkable recovery of the immune system after treatment, or a relatively high resistance of the immune system to the present treatment.

Effect of maternal ethanol consumption on in vitro tumor necrosis factor, interleukin-6 and interleukin-2 production by rat milk and blood leukocytes.

It has been shown that the transfer of immunity via lactation plays an important role in providing early protection to the neonate. Maternal ethanol consumption also results in a reduced transfer of immunity to their neonates against a Trichinella spiralis infection. Because of the known presence of cytokines in milk and their important role in inflammation, we tested the effects of maternal ethanol consumption on cytokine production by milk and blood cells from T. spiralis-infected rats. With T. spiralis antigen, Concanavalin A (Con A) or lipopolysaccharide stimulation, milk cells from both ethanol-treated and pair-fed groups were capable of producing tumor necrosis factor (TNF), interleukin (IL)-6 and IL-2. There were no differences between groups for TNF or IL-6 production by milk cells. Milk cells from the ethanol group produced a significantly higher amount of IL-2 upon Con A stimulation, as compared with that from the pair-fed group (16 +/- 4 units/10(6) cells vs. 4 +/- 1 units/10(6) cells, p < 0.05). After stimulation with Con A, blood cells from the ethanol group produced significantly lower amounts of TNF (17 +/- 15 units/10(6) cells) than that from the pair-fed group (102 +/- 64 units/10(6) cells, p < 0.05). The amount of TNF and IL-6 produced by milk cells was significantly lower, as compared with that produced by blood cells. This study suggests that ethanol consumption has some modulatory effects on cytokine production by milk and blood cells.

Dynamics of rat muscle mitochondria uncoupling in trichinellosis

Wandurska-Nowak E. and Boczon K. 1993. Dynamics of rat muscle mitochondria uncoupling in trichinellosis. International Journal for Parasitology 23: 587–590. Parallel oxygraphic measurements of respiratory control index (RCI) and spectroscopic measurements of the activity of two mitochondrial enzymes from rat muscle, mtATPase and succinate dehydrogenase were performed. In Trichinella spiralis infection, in comparison with Trichinella pseudospiralis infection, a delayed stimulation of mtATPase in 3rd week p.i. was observed and the degree of stimulation was weaker. A second peak of mtATPase stimulation appeared at the beginning of the encystation phase. The two-phase phenomenon was accompanied by a sharp decrease of RCI to 39% of normal values and almost 2-fold increase in the ratio of Mg 2+-stimulated to Mg 2+ -unstimulated mtATPase. By the end of investigations (7th week) the level of SDH normalized, but mtATPase was still elevated. The results, as in the case of T. pseudospiralis infection, support the testing of mtATPase activity to follow bioenergetic changes in tissue parasite infections. In comparison with oxygraphic measurements ATPase testing needs a much smaller amount of biological material and ATP.

Occurrence and characteristics of hypodense eosinophils in rats infected with Trichinella spiralis.

Hypodense eosinophils are observed in peripheral blood and tissue from patients with eosinophilia due to helminthic infections. In this study, the variation in eosinophil density was examined in rats during Trichinella spiralis infection. Hypodense eosinophils were observed in the peripheral blood in association with Trichinella infection. In peritoneal fluid, which was representative of tissue fluid, a majority of eosinophils were hypodense regardless of the infection. During the course of tissue eosinophilia after the infection, there was a particular increase in the lowest dense cells in the population of hypodense eosinophils. Eosinophils with lower density, demonstrated larger diameters and had more potent cytotoxic activity.

Influence of resistant and susceptible genotype, IL-1, and lymphoid organ on Trichinella spiralis-induced cytokine secretion.

The relative importance of cell-mediated inflammatory responses and antibody-mediated responses in controlling parasitic helminth infection is debated. To study the relationship between these responses and resistance or susceptibility to primary Trichinella spiralis infection, we infected resistant AKR mice and susceptible B10.BR mice and analyzed the lymphokines IL-2, IFN-gamma, and IL-5 produced by their T cells as a function of time and lymphoid organ. IL-2-secretors occurred maximally between days 3 and 6 postinfection, whereas IL-5-secretors peaked between days 6 and 9. Previously, we found that IFN-gamma producers peaked before day 6, whereas IL-4 producers peaked between days 6 and 9. Most cytokine secretors were CD4+. The simultaneous development of IL-2- and IFN-gamma-secreting cells, and IL-4- and IL-5-secreting cells, suggests that the infection may be stimulating T cells to differentiate into cells capable of secreting specific cytokine sets, analogous to the postulated Th1 and Th2 subsets. In the spleen and mesenteric lymph nodes, cells from B10.BR mice secreted more IL-5 than cells from AKR mice, as we found previously for IL-4. For both strains, mesenteric lymph node cells produced more IL-5 than splenocytes. The AKR mesenteric lymph node cells produced more IL-2 than the B10.BR cells, but the reverse occurred in splenocytes. The AKR peripheral lymph node cells also secreted more IFN-gamma than the B10.BR cells, but the strains were equivalent for peritoneal exudate cell IFN-gamma production. Thus, the lymphoid organ microenvironment plays an important role in regulating cytokine-secreting cell development in this system. We also tested the possible regulatory role of IL-1. Exogenous rIL-1 alpha increased IFN-gamma secretion early but not late in mesenteric lymph node cells from both strains; this reflected an increased IFN-gamma-secreting cell frequency, not a change in IFN-gamma mRNA transcript level. Exogenous rIL-1 alpha did not consistently affect IL-2, IL-4, or IL-5 secretion. These data suggest that IL-1 alpha availability in vivo may regulate IFN-gamma-secreting cell development. In sum, early activation of IFN-gamma-secreting T cells in lymph nodes, with little subsequent activation of IL-4- and IL-5-secreting cells, distinguished the resistant from susceptible strain responses to T. spiralis infection, and IL-1 alpha and lymphoid organ environment influence IFN-gamma-secreting cell activation.

IFN-gamma- and IL-5-producing cells compartmentalize to different lymphoid organs in Trichinella spiralis-infected mice.

The differential induction of cytokines associated with Th1 and Th2 subsets has recently been described during Trichinella spiralis infection. Increased levels of resistance appear to correlate with elevated levels of the Th1-associated cytokines, IFN-gamma and IL-2. In the present report, a filter immunoplaque assay is used to quantify the actual numbers of cells that secrete IFN-gamma and IL-5. It is demonstrated that, in T. spiralis-infected B10.Q mice, Th1- and Th2-associated responses are compartmentalized to different lymphoid organs. Thus, Ag-induced IFN-gamma-producing cells predominate in the spleen, whereas IL-5-producing cells prevail in the mesenteric lymph nodes (MLN). A corresponding compartmentalization of Ag-specific IgA and IgG1 antibody-secreting cells to the MLN is also noted. The virtual absence of Th1-associated responses in the MLN appears to be an Ag-associated phenomenon. MLN from either naive or T. spiralis-infected mice do have the capacity to secrete IFN-gamma if stimulated with Con A. The striking compartmentalization of Ag-driven cytokine responses seen in this parasite system may facilitate study of the mechanisms that regulate the induction of Th1 and Th2 subsets.

T-lymphocyte modulation of intestinal muscle function in the Trichinella-infected rat

Jejunal longitudinal muscle from Trichinella-infected Sprague-Dawley rats generates increased tension in vitro 6 days after infection. To investigate the extent to which this increase is dependent on T lymphocytes, smooth muscle contraction was examined in athymic rats (rnu/rnu) and in their euthymic (rnu/+) littermates 6 days after infection. In both thymus-bearing and athymic rats, the highest concentrations of Trichinella were found in the jejunum where mucosal myeloperoxidase activity was increased. Muscle from infected euthymic rats generated more tension to carbachol or 5-hydroxytryptamine than that of noninfected controls. In contrast, there was no difference between the responses of muscle from infected and control athymic animals. However, when athymic rats were reconstituted with splenic mononuclear cells, increased tension to carbachol or 5-hydroxytryptamine was observed after infection. T-lymphocyte activity was demonstrated by in vitro assays in euthymic or reconstituted rats but not in athymic rats. It is concluded that some changes in intestinal smooth muscle function following Trichinella spiralis infection in the rat are T lymphocyte dependent. These results support the concept of immunomodulation of intestinal smooth muscle function.

A Dot-ELISA Mimicry Western Blot Test for the Detection of Swine Trichinellosis

A dot enzyme-linked immunosorbent assay (dot-ELISA) using antigens purified by monoclonal antibody affinity chromatography was developed for detecting Trichinella spiralis infection in swine. The test was as sensitive as an ELISA using excretory-secretory products as antigen and western blot analysis, and nearly as specific as the western blot. The dot-ELISA detected all of 20 low infections (0.08-4.74 larvae per gram of diaphragm), most of them by 5-6 wk postinfection. Sera from 1,960 farm-reared swine were tested by conventional ELISA, dot-ELISA, and western blot. Of the 1,960 sera, 262 (13.4%) were considered positive on conventional ELISA, 16 (0.82%) by dot-ELISA, and 15 (0.77%) by western blot. The improved specificity was achieved by employing species-specific denatured antigens. More importantly, the dot-ELISA was much simpler to perform than western blot analysis. The principles employed in this test can be adapted to other infectious diseases, such as AIDS.

Peripheral blood mononuclear cell subsets during Trichinella spiralis infection in pigs

The immune response of 'Yugoslav meat breed' pigs inoculated with low doses of Trichinella spiralis muscle larvae was followed over two to nine weeks of primary infection, by analysing changes in peripheral blood mononuclear cell subsets, the development of a humoral antibody response and muscle larvae burden. During the course of the infection, infected animals showed a persistent elevation of both CD4+ and CD8+ T cell subsets from days 15 to 60 after the parasite exposure. During this time, the number of peripheral blood mononuclear cells expressing major histocompatibility complex class II antigens was also increased, while no significant differences were found in the number of circulating monocytes/macrophages and B cells over time. Humoral antibody responses to muscle larvae excretory-secretory products were evident as early as 41 days after infection, while the muscle larvae were recovered as early as 27 days after infection. The increased levels of CD4+ and CD8+ T cell subsets, as well as cells expressing major histocompatibility complex class II antigens in pigs exposed to T spiralis, may be indicative of some considerable alterations in cell subsets that are involved in the regulation of the swine immune response to this parasite.

A Spectrum of Antibody Response with Time after Trichinella spiralis Infection in Rats

A chronology of class-specific antibody response against the Trichinella spiralis infection in Fischer rats was investigated. G-class antibodies against the cuticle inner layer(s), hypodermis, hemolymph, glycogen aggregates, discrete areas in genital primordial cells, intestinal gland cell granules, and cytoplasmic granules in the cords were detectable 2 wk after infection (the rapid-responding group), whereas G-class antibodies against the cuticle surface, stichocyte granules, and the esophagus-occupying substance were detected 6 wk after infection (the slow-responding group). M-class antibodies recognized a narrower spectrum of antigens than did G-class antibodies; M-class antibodies against hemolymph, cord granules, and intestinal gland cell granules were not detectable. M-class antibodies tended to decrease in titer with time after infection. This tendency was more striking with antibodies against the rapid-responding group than with those against the slow-responding group. This information sheds light upon antibody response against many antigenic components of T. spiralis muscle larvae.

The Effects of Ethanol Consumption on the Expression of Immunity to Trichinella spiralis in Rats

The effects of ethanol ingestion on immune responses of female rats against Trichinella spiralis (T. spiralis) infections were investigated. Female rats were pair-fed either ethanol-containing or isocaloric control liquid diets for 68 days, during which time they underwent one pregnancy cycle. For some studies, animals were fed ethanol diets for 68 days beginning prior to pregnancy and continuing through lactation and involution. They were infected on Day 7 of involution with 1000 L1 larvae of T. spiralis and tested for a primary rejection response 10 days later. To test for a secondary immune response, rats were infected with T. spiralis, placed on ethanol-containing liquid diet 15 days later, and after 68 days on diets, challenged with 1000 T. spiralis larvae and killed 3 days later. For primary immunized studies, ethanol-treated animals demonstrated significantly lower levels of anti-T. spiralis serum antibodies in ELISA, reduced rates of H3 thymidine incorporation by lymph node cells stimulated with T. spiralis antigen and significantly higher numbers of intestinal worm burdens (decreased immunity) compared with pair-fed controls. For animals sensitized to T. spiralis prior to pregnancy and given a secondary challenge during involution, no differences were found between ethanol and pair-fed animals in their ability to reject their worm burdens or in anti-T. spiralis serum antibody levels; however, ethanol-treated animals showed reduced rates of thymidine incorporation by lymph node cells when stimulated with T. spiralis antigen.(ABSTRACT TRUNCATED AT 250 WORDS)