Alteration of cellular immune responses by nutrition and weaning in calves

The effects of two levels of nutrition (400 g and 1000 g air dry matter milk substitute powder per day) and three ages of weaning (rive, nine and 13 weeks) on cellular immune responses were determined in 32 calves. The lower level of nutrition was found to increase skin sensitivity responses to keyhole limpet haemocyanin (KLH) and decrease lymphocyte blastogenesis test (LBT) responses to ConA and pokewood mitogen (P<0·05). Weaning at five weeks old resulted in increased KLH skin responses at nine weeks old compared with unweaned calves and decreased LBT responses to ConA and phyto-haemagglutinin at 10 weeks old compared with calves weaned at nine weeks old (P<0·05). Weaning at five weeks old also increased peripheral blood concentrations of BoCD2+ and BoCD8+ lymphocytes (P<0·05). The results show that the choice of husbandry conditions alters cellular immune responses in young calves and suggest that early weaning effects are essentially nutritional.     

Immune responses of infected and vaccinated Hereford cattle to antigens of the cattle tick, Boophilus microplus

Responses of infested and vaccinated Hereford cattle to Boophilus microplus antigens were measured by enzyme-linked immunosorbent assay (ELISA), lymphocyte blastogenesis assay (LBA) and intradermal skin tests. Responses against soluble salivary gland extracts (SGS), salivary gland membrane (SGM), soluble gut extracts (GS), gut membrane (GM), soluble larval extracts (LS) and larval membrane (LM) antigens were tested. In one experiment, cattle infested with up to 160,000 ticks had positive cellular responses to SGS and significant antibodies against LM, GM, SGM, and SGS. Cellular responses to Concanavalin A were not depressed following infestation. Cattle vaccinated with GM, using Quil A as adjuvant, had positive cellular responses to gut and salivary gland antigens and significant antibody responses to all antigens tested. The antibody levels of vaccinated cattle were significantly higher than the antibody levels of infested cattle (P less than 0.05). In a second experiment, immune responses of cattle infested with 40,000 ticks were studied during 38 days. Cellular responses in LBA to several tick antigens were transiently elevated and significant levels of antibody were measured against LM, GM, SGM and SGS, from day 25 (P less than 0.05). Infested cattle had positive skin reactions following intradermal injection of larval and adult tick antigens (P less than 0.05).     

Immune modulation by Ostertagia ostertagi and the effects of diet

IgG1 antibody responses to Ostertagia ostertagi third stage larvae (L3) and the third party antigen, keyhole limpet haemocyanin (KLH), and faecal egg counts were determined in calves infected with a single dose of O. ostertagi and in uninfected, pair-fed calves. The infected and uninfected calves were given diets either high (H) or low (L) in protein and energy. The diets were within the normal range of husbandry practice in the UK. IgG1 antibody responses to L3 antigen were significantly greater from 6 weeks post-infection in infected calves given the L diet than in infected calves given the H diet (P less than 0.05). The effects of diet and infection on anti-KLH IgG1 responses were independent of each other. IgG1 responses to KLH were decreased by infection and by the L diet compared with the H diet.     

Experimentally induced Cooperia oncophora infection in calves: lymphocyte blastogenic and delayed hypersensitivity responses

Lymphocytic responses in peripheral blood and visceral lymph to Cooperia oncophora antigen and skin tests were determined in 35 Holstein male calves that were inoculated orally with single or multiple doses of C oncophora infective larvae. Several calves were vaccinated or given immune serum before larvae were inoculated. Antigen-specific in vitro blastogenesis of blood and lymph lymphocytes and delayed-type hypersensitivity reactions were observed in several inoculated, vaccinated, and/or passively immunized calves. Most calves that had delayed skin reactions also had in vitro lymphocyte responses to C oncophora antigen. The lymphocyte and skin responses were inconsistent and variable in time of onset--the earliest lymphocyte response occurring 7 days after calves were inoculated. A cellular immune response was induced by both dermal vaccination and oral inoculation; however, passive immunization by IV administration of immune serum simultaneously with inoculation did not have an apparent effect on the cellular response, as measured by the lymphocyte blastogenesis test or dermal testing. Although cellular immune responses were observed in several calves infected with C oncophora, there was no apparent relationship between the specific responses and number of nematodes establishing infection in calves after either single- or multiple-dose oral inoculations.     

Evaluation of relationship between plasma fibrinogen concentration and tuberculin testing in red deer

After intradermal injection of bovine purified derivative (PPD), increases in plasma fibrinogen concentration and plasma viscosity developed in red deer (Cervus elaphus) with a history of tuberculosis caused by Mycobacterium bovis. Serum haptoglobin concentrations were also found to increase under similar circumstances. The increases were reproducible and did not appear to be related to mustering, stress, or the handling associated with injection of PPD. A significant (P less than 0.05) direct relationship was found between the increase in plasma fibrinogen concentration and various markers of bovine tuberculosis infection, such as stimulation of lymphocyte transformation in response to bovine PPD and the diameter of intradermal tuberculin skin test reactions. A stronger correlation (P less than 0.01) was found with the volume of intradermal tuberculin skin test reactivity, and the strongest correlation (P less than 0.001) was with the presence of circulating antibovine PPD antibody.     

Adverse immune reactions and the pathogenesis of Ostertagia ostertagi infections in calves

The possible development of type-1 hypersensitivity reactions in the abomasal mucosa caused by soluble L3 products of Ostertagia ostertagi was studied in 4-month-old calves sensitized by repeated exposure to L3 over a 50-day period followed by anthelmintic treatment. Four groups each of 4 calves were used. Group 1 served as nonsensitized controls and group 2 as sensitized controls, group 3 was challenge exposed at 2-week intervals beginning at week 10 with a soluble L3 product (OAG), and group 4 was challenge exposed at 2-week intervals with an oral dose of L3, followed by anthelmintic treatment 3 days later. All calves infected with L3 became sensitized, as indicated by a positive reaction to an intradermal skin test. However, a passive cutaneous anaphylaxis was only partly effective in indicating the presence of homocytotropic antibody in the infected calves. Sensitized calves had significantly (P less than 0.05) higher eosinophil counts and plasma pepsinogen values for the entire 14 weeks than uninfected controls. Globule leukocyte and mast cell counts from the abomasal mucosa were also significantly (P less than 0.05) higher. Studies for possible immunomodulation revealed that lymphocyte counts decreased between every 2-week challenge-exposure period for groups-3 and -4 calves. A transient depression of blood lymphocyte (BL) responses to phytohemagglutinin (PHA), a T-cell mitogen, was observed over the first 8 weeks in the infected calves. Increases in BL responses to OAG were also observed. Differences were not observed in BL responses to pokeweed mitogen, a T- and B-cell mitogen. Blood lymphocyte responses to PHA in group-3 calves were low following the initial challenge exposure with OAG.(ABSTRACT TRUNCATED AT 250 WORDS)     

Onset,kinetics and duration of in vivo and in vitro delayed hypersensitivity responses of neonatal calves sensitized with mycobacterial components

Newborn calves developed delayed skin-test responses 10 days after a single intradermal inoculation at birth with 150 μg of purified derivative of Mycobacterium bovis associated with 150 μg of mycobacterial immunopotentiating glycolipid P3 and oil droplets. Development of tuberculin skin-test reactivity was detected simultaneously with increased lymphocyte transformation responses. Longitudinal studies revealed that two of three calves tested at 17 months of age were still skin-test positive. The kinetics of cellular hypersensitivity responses in neonatal calves sensitized with mycobacterial components are compared to the results of acquired cellular resistance and delayed hypersensitivity studies in phylogenetically disparate species. We concluded that bovine neonates have the capacity to rapidly develop cellular immune responses following stimulation with mycobacterial antigens.     

Effect of transcutaneous immunization with co-administered antigen and cholera toxin on systemic and mucosal antibody responses in sheep

Direct application of antigens to skin together with an adjuvant, a procedure called transcutaneous immunization (TCI), can induce systemic immune responses in mice, humans, cats and dogs. In previous studies we found that cholera toxin (CT) applied topically on unbroken skin induces systemic antibody and lymphocyte proliferative responses in sheep. The current study examined whether concurrent administration of CT and tetanus toxoid (TT) delivered transcutaneously could induce specific antibody responses to both antigens in sheep. Antibodies to both TT and CT were induced by TCI although antibody titres in serum to TT were higher in sheep receiving TT plus alum by intramuscular injection (n=5) than TT plus CT by TCI (n=5). The ratio of IgG1/IgG2 antibody to TT in serum was near unity, and the route of immunization, TCI versus injection, did not influence this ratio. In contrast, the ratio of IgG1/IgG2 antibody differed significantly between the two antigens, TT and CT, delivered by TCI, with a higher proportion of IgG1 antibody in serum to CT than TT. Antibody to TT was detected in lung washes from TCI and injection groups, with IgG1 predominating over IgG2 in both groups. IgA antibodies to CT and TT were detected in sera of CT and TT-immunized groups respectively but in lung washes IgA antibody to TT was detected only in the injection group. Results show that TCI induced systemic antibody responses to CT and the co-administered antigen TT, whereas no evidence was obtained for mucosal IgA responses following TCI.     

Humoral response of cattle to aerosolized Micropolyspora faeni

Calves (7) were exposed to antigens of Micropolyspora faeni by the aerosol route for 9 weeks. The humoral immune response of calves to M faeni antigens was studied; immunoglobulins (Ig) E, G1, G2, A, and M were measured weekly in serum and nasal secretions by enzyme-linked immunosorbent assay (ELISA). Intradermal injection of antigen was performed during the 6th and 9th weeks; responses were evaluated at 30 minutes, 6 to 8 hours, 24, and 48 hours after injection. Total IgE levels in serum and nasal secretions, evaluated weekly, did not show any elevation. Micropolyspora faeni-specific IgE, IgA, IgG1, and IgG2, but not IgM, were produced by calves exposed to the antigen by the aerosol route; individual variability in magnitude of the response was marked. Thirty-minute skin tests were positive for cytotropic antibody in 2 of 3 aerosol-exposed calves by the 9th week, but delayed-type reactivity was not present. The ELISA test results were compared with those from sera of saline solution aerosol-exposed calves and from a parenterally immunized calf. Comparison of isotype-specific ELISA results obtained from M faeni aerosol-exposed calves with ELISA results from calves exposed to aerosolized ovalbumin according to a similar procedure indicated inherent problems in evaluating immune responses to environmental antigens. Aerosolized M faeni elicited a substantial antibody response. In particular, it is noteworthy that antigen-specific IgE responses were detected.     

Immunity of specific pathogen-free lambs to challenge with an aerosol of Pasteurella haemolytica biotype A serotype 2. Pulmonary antibody and cell responses to primary and secondary infections

Specific pathogen-free (SPF) lambs previously exposed to an aerosol of P. haemolytica biotype A serotype 2 (A2) were immune to subsequent challenge with an aerosol of P. haemolytica A2. Untreated control lambs were not immune to this challenge. The local immune responses of the lung to these challenges were examined. High IgG and IgA titres to P. haemolytica and high levels of opsonizing antibody against P. haemolytica were present in the lung washings from previously infected immune lambs at autopsy, seven days after the second infection. Lung washings from control lambs, 7 days after challenge with P13 virus and P. haemolytica A2, had no IgG titres, very little opsonizing activity but did have IgA titres which were significantly higher than in unchallenged control lambs. The cellular response of animals challenged with P13 virus and P. haemolytica was significantly greater than that of unchallenged controls or of lambs exposed only to P. haemolytica. However, this finding was complicated by the response to P13 virus. Lymphocytes from lung washings of all lambs failed to respond in a lymphocyte stimulation test to phytohaemagglutinin while blood lymphocytes did respond. There was little specific response to P. haemolytica antigen in the test.     

Induction of systemic immune responses in sheep by topical application of cholera toxin to skin

Cholera (and related) toxins (CT) when applied topically on unbroken skin induce systemic immune responses in mice, a procedure called transcutaneous immunization (TCI). The current study examined the capacity for TCI to induce systemic immune responses in sheep. Three groups (n=5 per group) were immunized at day 0 (priming) and day 28 (boosting) with 250 microg of CT in water by TCI, with 25 microg of CT in alum by intramuscular injection, or not immunized. Serum samples were taken at days 0, 28, 42, 56 and 70 after immunization for measurement of CT-specific IgG as well as CT-specific IgG1, IgG2, IgA and IgM antibodies by ELISA. After immunization, IgG, IgG1 and IgG2 antibody in immunized groups were significantly higher than in the control group, and boosting further increased these titres. IgG, IgG1 and IgG2 in the injection group were significantly higher than in the TCI group. There was a preponderance of IgG1 antibody, relative to IgG2, in both immunized groups. CT-specific IgA and IgM were detected in both immunized groups. Lymphocyte proliferation to CT was measured at day 90. A CT-specific lymphocyte proliferative response (stimulation index>2) was detected in all sheep from the injection group, in two sheep from the TCI group and in none of the controls. Results demonstrated that TCI induces primary and secondary antibody responses and specific proliferative responses to CT in sheep.     

Direct bovine leukocyte migration inhibition assay: standardization and comparison with skin testing

A direct leukocyte migration inhibition (LMI) assay under agarose was standardized for use in cattle. The procedure included maintenance of culture conditions at pH 7.4 in a humidified, 39 C incubator with 98% air and 2% CO2. Optimum demonstration of incubation was obtained using a 9-hour incubation period. The LMI assay was used to detect reactivity to keyhold limpet hemocyanin (KLH) and horseshoe crab hemocyanin (HCH). Calves sensitized to KLH had specific reactivity for KLH and occasional reactivity for HCH. Similarly, calves sensitized to HCH showed reactivity for HCH and occasional reactivity for KLH. The nonspecific reactivity in the LMI was more frequent after skin testing with both antigens. Specific association of LMI reactivity and DTH reactivity was found in 8 of 10 animals. There were no nonspecific skin test responses. The results suggest that consecutive LMI assays are required to establish trends in reactivity and these should be used in conjunction with other in vitro assays of cellular immune reactivity.     

Effect of transportation and weaning on humoral immune responses of calves

Transportation exposes cattle to stress and results in increased morbidity and mortality. An investigation was made of the effects of transport and another important stressor, weaning, on the immune function of calves by determining Immoral immune responses to keyhole limpet haemocyanin (KLH). In a 2 × 2 factorial designed experiment, suckled calves were either (1) weaned at housing (day 0) and not transported, (2) weaned at housing and transported, (3) weaned while still at pasture nine to 13 days prior to housing and not transported or (4) weaned at pasture and transported. All calves were immunized with KLH at housing (day 0) and serum samples were collected subsequently to determine class and subclass anti-KLH antibody responses (IgG₁, IgG₂, IgA and IgM) by direct elisa. Increased anti-KLH IgG₁ and IgG₂ concentrations were shown in calves that were weaned prior to housing and transported on day 10 (P<0·05 and P<0·01 respectively). Transported calves had increased IgGI concentrations on day 20 (P<0·05) compared with calves that were not transported. However, calves weaned at housing and not transported had increased IgA and IgM responses on day 30 compared with the other groups of calves (P<0·05). This study shows that transportation and weaning affect the Immoral immune responses of suckler calves and that the effects persist for several weeks. However, the effects of the treatments were not consistent for all antibody classes measured.     

Shipping suppresses lymphocyte blastogenic responses in Angus and Brahman X Angus feeder calves

An experiment using 40 Angus or Brahman X Angus preconditioned feeder calves was conducted to evaluate the influence of shipping on cellular immune reactivity. Steers were allotted on the basis of weight and breed to a control or shipped group. Shipped steers were trucked 700 km to a feedlot; control steers remained at the ranch of origin. Total and differential leukocyte counts, phytohemagglutinin skin-test responses, lymphocyte blastogenic responses, monocyte phagocytic function, packed cell volumes and concentrations of plasma cortisol were determined before, immediately after and 1 wk after shipment. At unloading, total leukocytes were increased (P less than .05) in shipped Angus steers. Shipped steers also had higher (P less than .01) numbers of neutrophils. Skin-test responses to phytohemagglutinin were higher (P less than .05) in Angus than in Brahman X Angus steers, but shipping did not influence the reaction. Lymphocyte blastogenic responses were lower (P less than .05) in shipped steers; however, cortisol levels in plasma were not elevated (P greater than .10) in shipped calves. Monocyte phagocytosis and packed cell volume were not influenced by shipping. These data suggest that shipped steers have suppressed lymphocyte blastogenic responses.     

Altered immune responses to a heterologous protein in ponies with heavy gastrointestinal parasite burdens.

This study was performed to test the hypothesis that immunity to heterologous vaccination would improve when the parasites were removed. It was also expected that parasitised ponies would exhibit a biased Th2 cytokine response to KLH immunisation. Helminth parasites are common in horses even in the era of highly effective broad-spectrum antiparasiticides. These parasites have been shown to alter the outcome to heterologous immunisation in a number of host species. The effect of gastrointestinal parasites on heterologous vaccination has not been addressed in equids. In the current study, humoral, lymphoproliferative, and cytokine responses to a single i.m. injection of keyhole limpet haemocyanin (KLH) were compared between groups of ponies with high, medium or low gastrointestinal parasite burdens. Antibody levels determined by ELISA showed that animals with low levels of parasites had a trend toward increased KLH specific total immunoglobulin, IgG(T) and IgA compared to heavily parasitised ponies. Medium and heavily parasitised ponies demonstrated a trend toward reduced lymphoproliferative response to KLH that was not restored after the addition of interleukin-2 (Il-2). Cells from these ponies also produced significantly lower levels of IL-4 compared to lightly parasitised ponies. These data indicate that heavily parasitised ponies have uniformly decreased cellular and humoral immune responses to soluble protein immunisation. The mechanisms involved may have potential deleterious effects on standard vaccine protocols of parasitised equines.     

Effects of cyclophosphamide on the lymphoid tissues and humoral and cellular immune responsiveness of young calves.

Cyclophosphamide (CY) was given IV to 5-month-old calves (ten doses; each dose of 5.0 mg/kg, 2-day intervals between doses). The effects of CY on circulating leukocytes, lymphoid tissues, and the humoral and cellular immune responses were assessed. The numbers of total leukocytes, lymphocytes, and neutrophils and platelets decreased significantly. The lymphocyte population was depleted in the cortex of the thymus and B-dependent areas of the spleen and lymph nodes. Significant decreases occurred in the frequency of the peripheral blood lymphocytes-bearing surface immunoglobulin (Ig) and in serum IgM and IgG concentrations. Primary serum antibody responses to avian erythrocytes and Brucella abortus strain 19 antigens were diminished or delayed. The blastogenic responses of peripheral blood lymphocytes to phytohemagglutinin P, concanavalin A, pokeweed mitogen, and to purified protein derivative and B abortus antigens were enhanced as was the delayed hypersensitivity reaction to the tuberculin skin test. While a diminished humoral immune response was associated with CY treatment, the cell-mediated response was potentiated. The effect of CY was transitory with most variables returning to near base line within 24 days after CY was ceased.     

Modulation by colostrum-acquired maternal antibodies of systemic and mucosal antibody responses to rotavirus in calves experimentally challenged with bovine rotavirus

The effect of colostral maternal antibodies (Abs), acquired via colostrum, on passive protection and development of systemic and mucosal immune responses against rotavirus was evaluated in neonatal calves. Colostrum-deprived (CD) calves, or calves receiving one dose of pooled control colostrum (CC) or immune colostrum (IC), containing an IgG1 titer to bovine rotavirus (BRV) of 1:16,384 or 1:262,144, respectively, were orally inoculated with 105.5 FFU of IND (P[5]G6) BRV at 2 days of age. Calves were monitored daily for diarrhea, virus shedding and anti-BRV Abs in feces by ELISA. Anti-rotavirus Ab titers in serum were evaluated weekly by isotype-specific ELISA and virus neutralization (VN). At 21 days post-inoculation (dpi), all animals were euthanized and the number of anti-BRV antibody secreting cells (ASC) in intestinal and systemic lymphoid tissues were evaluated by ELISPOT. After colostrum intake, IC calves had significantly higher IgG1 serum titers (GMT=28,526) than CC (GMT=1195) or CD calves (GMT<4). After BRV inoculation, all animals became infected with a mean duration of virus shedding between 6 and 10 days. However, IC calves had significantly fewer days of diarrhea (0.8 days) compared to CD and CC calves (11 and 7 days, respectively). In both groups receiving colostrum there was a delay in the onset of diarrhea and virus shedding associated with IgG1 in feces. In serum and feces, CD and CC calves had peak anti-BRV IgM titers at 7 dpi, but IgA and IgG1 responses were significantly lower in CC calves. Antibody titers detected in serum and feces were associated with circulation of ASC of the same isotype in blood. The IC calves had only an IgM response in feces. At 21 dpi, anti-BRV ASC responses were observed in all analyzed tissues of the three groups, except bone marrow. The intestine was the main site of ASC response against BRV and highest IgA ASC numbers. There was an inverse relationship between passive IgG1 titers and magnitude of ASC responses, with fewer IgG1 ASC in CC calves and significantly lower ASC numbers of all isotypes in IC calves. Thus, passive anti-BRV IgG1 negatively affects active immune responses in a dose-dependent manner. In ileal Peyer's patches, IgM ASC predominated in calves receiving colostrum; IgG1 ASC predominated in CD calves. The presence in IC calves of IgG1 in feces in the absence of an IgG1 ASC response is consistent with the transfer of serum IgG1 back into the gut contributing to the protection of the intestinal mucosa.     

Phenotypic comparison of ileal intraepithelial lymphocyte populations of suckling and weaned calves

Ileal intraepithelial lymphocyte (IEL) suspensions from suckling calves (1-3 weeks old) and weaned calves (3-6 months old) were phenotyped to determine whether there were differences in the lymphocyte populations consistent with postnatal maturation of the mucosal immune system. Flow cytometric comparisons of IEL from the two age groups revealed the presence of significantly larger proportions of CD4+ T lymphocytes and CD8+ T cells in the weaned animals. In contrast, there was a significantly larger proportion of B-B2+ IEL in the suckling calves. Freshly isolated IEL from both groups of calves expressed mRNA for TNF-alpha and IFN-gamma, but not IL-4 or IL-10. The B-B2+ IEL population was more closely examined by flow cytometry. These cells co-expressed IgM and CD21. However, they did not express IgA, IgG1, nor any of several additional leukocyte differentiation molecules. Immunohistochemical data confirmed the presence of IgM+ lymphocytes, and the paucity of IgA+ and IgG1+ lymphocytes in suckling calf ileum. However, substantial numbers of IgA+ and IgG1+ cells were observed in weaned calf ileum. Together, the data are consistent with ongoing postnatal maturation of the gut mucosal immune system.     

Interdigital skin test for evaluation of delayed hypersensitivity and cutaneous basophil hypersensitivity in young chickens

A skin test to assess T-cell mediated delayed hypersensitivity (DH) and cutaneous basophil hypersensitivity (CBH) was evaluated in the interdigital skin of young chickens. Three-day-old chickens were sensitized with Mycobacterium tuberculosis, and the DH reaction was elicited in the interdigital skin in 10-, 17-, 24-, and 31-day-old chickens by intradermal injection of tuberculin. Cutaneous basophil hypersensitivity was elicited in the interdigital skin of 10- and 14-day-old chickens by a single intradermal injection of phytohemagglutinin-P (200 micrograms). The effect of immunosuppression on the results of interdigital skin test for DH and for CBH was evaluated in chickens that were treated with dexamethasone daily for 4 days before testing. The DH reaction, as indicated by a significant (P less than 0.01) increase in the mean interdigital skin thickness, was detectable in 10-day-old chickens and was consistently evident in 17-, 24-, and 31-day-old chickens. The DH response in the interdigital skin of 24-day-old chickens was comparable with that elicited in the standard wattle test. The CBH reaction, as indicted by a significant increase (P less than 0.005) in skin thickness, was evident in the interdigital skin of 10- and 14-day-old chickens. Treatment with dexamethasone significantly decreased (P less than 0.01) the DH and CBH reactions. Results of the study indicated that the interdigital skin test may be used to evaluate normal and suppressed cell-mediated DH and CBH reactions in chickens as young as 10 and 14 days old.     

Protection from parasite-induced weight loss by the vaccination of calves with excretory-secretory products of larval Oesophagostomum radiatum

Excretory-secretory products (ESP) were collected from in vitro maintained Oesophagostomum radiatum larvae during the period in which the larvae molt from the third to fourth larval stage. The ESP were used to immunize 13 uninfected calves which were subsequently challenged with 1.7 X 10(4) infective O. radiatum larvae. Worm recoveries from immunized calves were reduced 23% compared to 12 unimmunized controls, while the number of intestinal nodules was 72% greater compared to unimmunized controls; however, neither difference was statistically significant. Immunized calves had enhanced serum IgG and IgA anti-ESP antibody responses upon challenge. No differences in serum IgG2 or IgM antibody or cellular immune responsiveness, as determined by in vitro antigen-induced lymphocyte proliferation, were seen. Vaccination with ESP did significantly protect calves from the weight loss seen in non-immunized calves. Unimmunized calves ceased gaining weight approximately 5 weeks after challenge and by 10 weeks after challenge had lost an average of 4 lbs. per calf. Over the same time interval (i.e., 5-10 weeks after challenge), the immunized calves gained an average of 23 lbs. per calf. These results strongly suggest that vaccination with ESP conferred an advantage to calves that is not correlated solely with the number of worms developing from challenge infection.