Humoral Immune Responses In Calves Research Articles

Protective efficacy of a recombinant adenovirus expressing novel dual F and HN proteins of bovine parainfluenza virus type 3

Bovine parainfluenza virus type 3 (BPIV3) is a viral respiratory pathogen that infects cattle and causes significant economic losses. We generated a recombinant adenovirus called rHAd5-F + HN by expressing the fusion (F) and hemagglutinin-neuraminidase (HN) glycoprotein of BPIV3 using the human adenovirus serotype 5 (rHAd5). We evaluated its effects on humoral and cellular immune responses in mice (n = 45) and calves (n = 9). Serum antibody responses were assessed by enzyme-linked immunosorbent assay (ELISA), hemagglutination inhibition (HI), and neutralising antibodies (NAb). After boosting immunity with rHAd5-F + HN, mice produced significantly higher levels of antibodies against the BPIV3 genotype A and genotype C strains. The production of antibodies exceeded those produced by adenoviruses rHAd5-F and rHAd5-HN, which express the F and HN glycoprotein, respectively. The percentages of splenic CD3+/CD8+T lymphocytes and IL-4+ cytokines in rHAd5-F + HN mice were considerably higher than those in the control group. Mice immunised with rHAd5-F + HN exhibited much lower viral loads in the lungs and tracheas compared to the control group. Additionally, the lungs of mice vaccinated with rHAd5-F + HN showed no notable histopathological changes. On the other hand, rHAd5-F + HN produced a humoral immune response in calves. Following the booster intramuscular injection with the rHAd5-F + HN, the serum antibody levels against BPIV3 genotype C strain were 1:20 452, 1:1024, and 1:426 in calves, as detected by ELISA, HI, and NAb, respectively. The HI and NAb levels against the BPIV3 genotype A strain were 1:213 and 1:85 in calves, respectively. These results indicate that rHAd5-F + HN effectively induced immunity against BPIV3 infection.

Humoral Immune Response in Calves Vaccinated with Monovalent Vaccines or a Trivalent Combination Thereof and Matching of These Vaccines to the Selected Circulating Foot-and-Mouth Disease Viruses in Ethiopia.

Foot-and-mouth disease (FMD) is an endemic, highly contagious, and devastating disease of livestock production in Ethiopia. Control of this disease relies mainly on prophylactic vaccination by willing farmers without a countrywide vaccination program. The objectives of this study were to quantify the humoral immune response and evaluation of the serological relationship of the vaccine strain used with representative field strain isolates. This was performed by primo vaccination of 6-9-month-old Holstein Friesian calves (35 treatment and 4 control calves) on day one and booster vaccination on day 28. Calves were vaccinated using the locally available National Veterinary Institute (NVI), Bishoftu, Ethiopia, inactivated aluminum hydroxide adjuvant monovalent (either O, A, SAT-2 alone) or trivalent (combination of A, O, SAT-2) vaccine (A/ETH/6/2000 (G-VII, O/ETH/38/2005(EA-3) and SAT-2/ETH/64/2009(XIII)). A 2 mL or 4 mL dose was used to vaccinate all calves except the animals that served as a control. In the case of the trivalent vaccine, a 4 mL dose was used to vaccinate calves. The serum was collected at 7, 14, 21, 28, and 56 days post-vaccination (d.p.v.). The humoral immune response was quantified by the solid-phase competitive enzyme-linked immunosorbent assay (SPC ELISA) and the virus-neutralization test (VNT). The serological relationship of heterologous and homologous viruses was also evaluated by adjuvant vaccine matching tests. The r1-value was determined using serum collected 21 d.p.v. An increase in immune response was observed from 7 d.p.v. to 28 d.p.v. in calves who received a 4 mL dose containing a 107.24 antigen load of 100 tissue culture infective dose (100TCID50) virus titer in the formulation. Upon receiving a booster dose on day 28, the humoral immune response was checked on the 56th day post-initial vaccination. Amounts of 54%, 72%, 79%, and 72% of inhibition for A, O, SAT-2, and trivalent vaccine in the three serotypes SPCE, respectively, was measured. Here, it was found that the immune response of calves increased from day 7 to 56, as evidenced by SPCE analysis. Likewise, an increase in antibody titer measured by a one-dimensional virus neutralization assay was also in line with SPCE analysis. This indicates that the vaccine is capable of inducing a neutralizing antibody that confers a protective immune response in 70%, 62%, and 100% heterologous field strains of A, O, and SAT-2 isolates, respectively, and has an average antigenic relationship of >0.3 with a standard deviation of +0.05 (N = 3) to the vaccine strains A/ETH/6/2000, O/ETH/38/2005 and SAT-2/ETH/64/2009, respectively, when using the one-dimensional virus neutralization test. The contribution and importance of this study is a confirmation of the vaccine and the field strain serological relationship for serotype SAT-2 strain and further research/change of vaccination strategy/ improvement in the currently used vaccine to cover a wide range of prevailing genotypes/lineages and induction of sound immune response after vaccination for serotype A and O strain. This study suggests that the trivalent vaccine produced by the National Veterinary Institute containing viral isolates from serotype O, A, and SAT-2 has a good serological relationship with the majority of circulating field strains in Ethiopia.

Dynamics of the humoral immune response of calves infected and re-infected with Cooperia punctata

The dynamics of the humoral immune response of calves were analysed after primary infection and re-infection with the intestinal nematode Cooperia punctata. 12 male 5 month-old Holstein-Friesian calves were randomly divided into two groups A and B. At the beginning of the experiment Group A animals were each infected experimentally with a single oral dose of 130,000 infective third stage larvae (L3) of C. punctata. The animals of Group B were kept as non-infected controls. The two calves from Group A with the highest infections died of cooperiosis at 32 and 44 days after infection (DAI), respectively. On DAI 100 the calves were treated with the recommended dose of oxfendazole. On DAI 180 the remaining four calves of Group A and three animals of Group B (B1) were infected with 260,000 L3 of C. punctata, while the other three calves of Group B (B2) served as non-infected controls. Monitoring of the humoral immune response predominantly demonstrated an IgG1 response against both adult and L3 antigen of C. punctata. Moreover, re-infections increased the levels of these immunoglobulins. IgA levels were less increased than IgG1 and no significant increase was observed in IgG2 and IgM levels. Immunoblotting analysis showed that total IgG present in the serum of the primary infected animals mainly reacted against adult proteins of 12–14 and 17–20 kDa and against L3 proteins of 33 and 43 kDa. After re-infection total IgG reacted with the same adult proteins but also with an adult 29 kDa protein.

Humoral immune response in calves to single-dose, trickle and challenge infections with Fasciola hepatica

In cattle experimentally infected with Fasciola hepatica, parasite specific IgG1 and IgG2 responses were studied. Additionally parasite specific IgE production was assessed by the Passive Cutaneous Anaphylaxis reaction. The primary infection was administered either as a single-dose or as a trickle infection over a 4-week period. Animals were challenged 4 months later. Titres of IgG1 and IgG2 against excretory–secretory parasite products (FhESAg), and against a whole-worm extract (FhSomAg) were measured by enzyme-linked immunosorbent assay (ELISA) in relation to weight gain, serum hepatic enzyme levels, and fluke infection rate. At necropsy, the mean number of flukes recovered was similar in both infected groups. The two ELISAs specific for bovine IgG1 showed analogous sensitivity and specificity (92% and 94%). Cross-reactivity was observed towards Echinococcus granulosus, Cysticercus tenuicollis, and C. ovis but not towards C. bovis, Cooperia spp., and Ostertagia spp. FhESAg gave rise to apparently more stable specific IgG1 titres as compared to FhSomAg. Mean IgG1 titres were significantly higher in the single-dose-infected group than in the trickle-infected group during the early migratory phase of the infection (week 2 to week 4 (FhSomAg) or week 6 (FhESAg)). IgG2 values were consistently lower than IgG1 levels. The kinetic response of both isotypes yielded a similar pattern. Specific IgE antibodies were detected in cattle of both infected groups from week 2 post-primary infection (PPI) onwards. The mean serum glutamate dehydrogenase (GLDH) and gamma-glutamyl transferase (γGT) activities were significantly higher in the single-dose-infected group for 3 weeks around peak levels (12–14 weeks PPI and 14–16 weeks PPI for GLDH and γGT respectively). Western blotting revealed a major antigenic fraction in FhESAg (26–30 kDa) recognized specifically by sera from F. hepatica infected calves as early as 6–8 weeks PPI. Experimental challenge caused no statistically significant modification of any parameter (IgG1 and IgG2 titres, enzymatic activities, immunoblotting) used to monitor the course of the infection. No correlation was found between fluke size and number, and antibody titres, suggesting that IgG1 production has little protective effect against F. hepatica infection.

Effects of Dietary Copper and Molybdenum on Copper Status, Cytokine Production, and Humoral Immune Response of Calves

Twenty-four male Holstein calves were used to determine the effects of dietary Cu and Mo on performance, Cu status, and immune function of calves. Calves were fed a milk replacer that was deficient in Cu for 8 wk and then were randomly assigned after weaning to one of four treatments: 1) control (no supplemental Cu or Mo), 2) 10mg of Cu (from CuSO4)/kg of dry matter (DM) (Cu diet), 3) 5mg of Mo (from Na2MoO4)/kg of DM (Mo diet), or 4) 5mg of Cu (from CuSO4) and 5mg of Mo (from Na2MoO4)/kg of DM (Cu + Mo diet). The basal diet was a semipurified diet that contained approximately 1.1mg of Cu and 1.1mg of Mo/kg of DM. Calves fed the Cu and Mo diets gained weight more efficiently than those fed the control and Cu + Mo diets during the 112-d study. By d 84 of the study, calves fed the Cu diet had higher plasma Cu concentrations and plasma ceruloplasmin activities than did calves fed the other three diets and had higher liver Cu concentrations on d 136. Plasma and liver Cu concentrations did not differ among calves fed the control, Mo, and Cu + Mo diets. At d 112, activity of Cu-Zn superoxide dismutase was lower in calves fed the Mo diet than in calves fed the Cu diet. Serum total antibodies to porcine erythrocytes (primary response) were lower in calves fed the Mo diet than in calves fed the Cu diet at 7, 14, and 21 d postinoculation. Production of tumor necrosis factor and interleukin-1 by isolated peripheral blood monocytes was not significantly affected by treatment. Although no differences were apparent in plasma or liver Cu concentrations among calves fed the control, Mo, and Cu + Mo diets, calves fed the Mo diet had a more severe Cu deficiency based on depressed humoral immune response and superoxide dismutase activity.

Recognition of low molecular weight Cooperia oncophora antigens after primary and trickle infection of calves with third-stage infective larvae

Recognition of low molecular protein fragments of adult Cooperia oncophora of calves was studied using sera obtained after a single oral (primary) infection with 100,000 infective 3rd-stage C. oncophora larvae, and after a 10-week period of trickle infection with moderate doses of these larvae. SDS-gel electrophoresis under reducing conditions, followed by western blotting, revealed that a complex of 14–16 kDa protein fragments, a 27 kDa and a 31.6 kDa band of Cooperia oncophora adult antigen were recognized by sera from calves both after primary and trickle infection. A 19 kDa and 30 kDa protein fragment were additionally bound by sera from calves after trickle infection. The data suggest that the systemic humoral immune response of calves to Cooperia oncophora after trickle infection is more pronounced (14–16, 27, 31.6 kDa) and extended (19, 30 kDa) than after primary infection with Cooperia.

Low molecular weight Cooperia oncophora antigens: Characterization and humoral immune responses in calves mono-infected with 100 000 infective larvae

Characteristics of the humoral immune response of Cooperia oncophora-infected calves to low molecular weight antigens of C. oncophora were studied. Immunoblotting with sera obtained from calves 6 weeks after a single oral infection with 100 000 third-stage (L 3) C. oncophora larvae revealed several corresponding antigenic fragments between adult worms and the fourth-stage (L 4) larvae. No reactivity in the immune sera was found against the L 3 stage. A previously defined complex of low molecular weight proteins (12–15 kDa) was found on both L 4 and adult Cooperia stages, but not on the L 3 stage. C. oncophora adults differed from the L 4 larvae at the 31 32 and 37 kDa level. Several adult and L 4 proteins were bound by biotinylated Concanavalin A, as was also true for L 3 proteins. A 31 32 kDa glycoproprotein of adult worms was recognized by a monoclonal antibody with specificity for phosphorylcholine. Using monoclonal antibodies in ELISA and Western blotting, the serum antibody response of C. oncophora-infected calves to adult worm antigen was almost entirely IgG1. Binding of the IgG1 antibodies was restricted to a complex of reduced 12–15 kDa protein(s) and a 27 kDa fragment of adult worms. The data suggest that the systemic humoral immune response of calves during a primary infection with C. oncophora consists mainly of an IgG1 response, and is directed to a non-glycoslyated Cooperia protein (molecular weight estimated at 12–15 kDa under reducing conditions and 18 kDa under nonreducing conditions). This protein is probably present in both L 4 larvae and adults. Since it was not bound by immune sera from calves mono-infected with several other nematodes, the 12–15 kDa protein complex may represent a Cooperia-specific component that can be used for serodiagnosis.

Influence of triiodothyronine injections on calf immune response to an infectious bovine rhinotracheitis virus challenge and nitrogen balance of lambs.

Three experiments were conducted to determine the influence of triiodothyronine (T3) or propylthiouracil (PTU) on the humoral immune response of calves challenged with infectious bovine rhinotracheitis virus (IBRV) and on nitrogen depletion and repletion of lambs deprived of feed and water for 3 d. In Exp. 1, 18 steer calves (BW 284 +/- 6 kg) challenged with IBRV were limit-fed (1.5% BW) a 60% concentrate diet and injected (s.c.) daily with alkaline saline, .4 mg of T3, or .8 mg of T3. Injections of T3 did not affect serum antibody titers to IBRV, blood leukocyte counts, or plasma free fatty acid, ceruloplasmin, and cholesterol concentrations but increased (P < .05) plasma glucose concentrations and decreased (P < .05) plasma urea N concentrations. In Exp. 2, 36 IBRV-challenged steers (BW 266 +/- 8 kg) were given ad libitum access to a 60% concentrate diet and injected (s.c.) daily with alkaline saline, .2 mg of T3, or .4 mg of T3. In contrast to Exp. 1, injections of T3 did not affect plasma glucose or urea N concentrations and reduced (P < .05) serum antibody titers to IBRV. In Exp. 3, eight wether lambs were limit-fed (600 g/d) a 36% concentrate pelleted diet and assigned to one of four treatments in a replicated 4 x 4 Latin square designed nutrient balance experiment involving periods of nutrient depletion and repletion. Treatments were as follows: 1) alkaline saline injection (s.c.), 2) 4 mg of PTU/kg BW in water, 3) .15 mg of T3 s.c. daily for 15 d, and 4) .15 mg of T3 s.c. daily for 7 d after the 3-d feed and water deprivation period. Thyroid status affected (P < .05) predeprivation N balance but did not affect N losses during the feed and water deprivation period. Retention of N during realimentation was affected (P < .05) by T3 treatment. Results of these experiments indicate that there is a complex interrelationship among stress, nutrient status, and thyroidal status and that the effects of T3 injections on immune and metabolic responses may be dependent on the nutritional status of the animal.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Dietary Selenium on the Primary and Secondary Immune Response in Calves Challenged with Infectious Bovine Rhinotracheitis Virus

The present study was conducted to determine if a selenium deficiency affects the primary and secondary humoral immune response of calves challenged with infectious bovine rhinotracheitis virus (IBRV) and to evaluate changes in glutathione peroxidase (GSH-Px) and creatine phosphokinase activities associated with selenium deficiency and viral stress. Treatments included 1) +Se (0.2 mg/kg diet) and 2) -Se (0.03 mg/kg diet). Calves were adapted to their assigned diets during an 84-d depletion period. By d 42 of the depletion period, whole-blood GSH-Px was lower for -Se calves than for +Se calves. Differences between treatments in plasma GSH-Px were observed by d 14. On d 84, Se-dependent GSH-Px activity in liver was higher in +Se calves. After depletion of -Se calves, animals in both treatments were intranasally inoculated with IBRV on d 0 and 35 of the 70-d challenge phase. Whole-blood and plasma GSH-Px increased after IBRV inoculation in +Se calves but not in Se-deficient calves. Serum IgM was higher for +Se calves throughout the challenge phase. Serum IgM increased in both treatments after primary IBRV inoculation but was unaffected by secondary challenge. Serum IgG was not affected by selenium status but increased in both treatments after secondary IBRV challenge. Serum IBRV antibody titers increased after inoculation on d 0 and 35. By d 49, IBRV antibody titers were higher for +Se calves than in Se-deficient calves. These results indicate that selenium deficiency may depress the immune response of calves challenged with a foreign pathogen.

Cellular and humoral immune responses in calves experimentally infected with salmonella dublin

Both humoral and cellular immune responses have been found to be operative in calves surviving smooth Salmonella dublin infection. Such calves showed rises in agglutinin titre and serum immunoglobulin as compared to uninfected controls. These calves at 3 weeks post infection showed a 3-4 fold increase of initial skin thickness at 5 h post antigen inoculation - an indication of immediate type of skin hypersensitivity reaction. The maximum increase in skin thickness of 2-3 fold was observed at 48 h post antigen inoculation - a criterion of delayed type of hypersensitivity reaction. There was a moderate increase, 28.1 per cent in the migration inhibition of peripheral leucocytes during the first week and a sharp increase, 45.7 and 66.6 per cent during the second and third week post infection, respectively. Passively immunized calves on challenge exhibited delayed clinical symptoms and mortality as compared to control animals, though the calves could not be protected against challenge. The serum immunoglobulin levels in passively immunized calves decreased moderately during the first week and markedly during the second week post challenge while in the control calves the levels dropped abruptly in the first week followed by death in both groups.