Assessment of T-dependent and T-independent immune responses in cattle using a B cell ELISPOT assay

Understanding the mechanisms that maintain protective antibody levels after immunisation is important for vaccine design. In this study, we have determined the kinetics of plasma and memory B cells detectable in the blood of cattle immunised with model T-dependent or T-independent antigens. Immunisation with the T-D antigen resulted in an expansion of TNP-specific plasma cells post-TNP primary and booster immunisations, which was associated with increased titres of TNP-specific IgG antibodies. Although no TNP-specific memory B cells were detected in the T-D group following the primary immunisation, we detected an increase in the number of TNP-specific memory B cells post-TNP boost. In contrast, no TNP-specific plasma or memory B cells were detected after primary or secondary immunisation with the T-I antigen. We then investigated if immunisation with a third party antigen (tetanus toxin fragment C, TTC) would result in a bystander stimulation and increase the number of TNP-specific plasma and memory B cells in the T-D and/or T-I group. TTC immunisation in the T-D group resulted in a small increase in the number of TNP-specific plasma cells post-TTC primary immunisation and boost, and in an increase in the number of TNP-specific memory B cells post-TTC boost. This bystander effect was not observed in the animals previously immunised with the T-I antigen. In conclusion, the present study characterised for the first time the B cell response in cattle to immunisation with T-D and T-I antigens and showed that bystander stimulation of an established T-D B cell memory response may occur in cattle.     

Different sources of dietary n-6 polyunsaturated fatty acids and their effects on antibody responses in chickens

1. Effects of linoleic and linolenic acid provided via different oil sources on total antibody (Ab) titres, Ab isotypes after primary and secondary immunisation, and cutaneous hypersensitivity (CH) responses to bovine serum albumin (BSA) and maleyl-BSA, respectively, were studied in pullets fed on one of 4 diets. The diets were the basal control diet enriched with either sunflower oil or safflower oil as sources of linoleic acid, and linseed oil as a source of linolenic acid, tested against a control diet supplemented with animal fat. 2. Total Ab and immunoglobulin (Ig) isotype responses to BSA were affected by diet after primary, and diet x immunisation effects after secondary immunisation. Higher total Ab and IgG titres to BSA were found especially after primary immunisation in birds given the sunflower oil enriched diet, whereas birds given sunflower oil mounted significantly lower IgM titres to BSA after primary and secondary immunisation. The antibody responses to maleyl-BSA were affected by diet after primary, and immunisation x diet interactions after secondary immunisation. Sunflower oil enhanced total and IgG Ab titres to maleyl-BSA after primary immunisation, but decreased IgM titres to maleyl-BSA after primary and secondary immunisation. Cutaneous hypersensitivity responses to BSA and maleyl-BSA were not affected by the diet. 3. It is concluded that modulation of the magnitude and isotype of Ab responses of poultry to T cell-dependent antigens is affected not only by type of essential fatty acids, but also by their source. In the present study the n-6 source, sunflower oil, showed strong enhancement of primary Ab responses, directed to both Th2 and Th1 antigens. On the other hand, the different effects of safflower oil imply that constituents other than n-6 acids within dietary plant oils may affect immune responsiveness. 4. The relationship between magnitude and isotype of Ab responsiveness, type of antigen, and essential fatty acids is discussed.     

Classical swine fever virus in plasma and peripheral blood mononuclear cells of acutely infected swine

The distribution of classical swine fever virus (CSFV) in plasma, monocytes, T and B lymphocytes in peripheral blood was monitored during experimentally induced acute classical swine fever infection in piglets. Six piglets were infected with 10(3.8) TCID50 of virus and blood samples taken up to 18 days post-inoculation (p.i.). Infectious virus was detected in monocytes, T and B lymphocytes to similar titres in five of the six infected piglets. Infectious virus was detected earlier in plasma than in any of the mononuclear cell subpopulations. No significant difference was observed in the period of time in which virus could be isolated from the three cell subpopulations. While a progressive lymphopenia developed, a marked B cell depletion was observed. However, B cells were apparently replaced by non-IgM-bearing mononuclear cells, as the proportion 'total lymphocyte/total leucocytes' remained unaltered throughout the experiment. Virus titres in plasma and peripheral blood mononuclear cells showed a tendency to increase as the disease progressed to its outcome.     

The ontogeny of humoral immunity in rainbow trout, Salmo gairdneri

The ontogeny of the humoral immune response to a 'thymus dependent' and a 'thymus independent' antigen, human gamma globulin (HGG) and Aeromonas salmonicida (AS) respectively, was investigated in the fry of rainbow trout, Salmo gairdneri, by direct immersion vaccination in the antigens (dose 5 mg/L HGG; 10(8) cells/ml AS; 30 minutes) at known ages/weights from 7 days post hatch, and at 1, 2, 3, and 4 months post hatch. Half the fry in each group were tested for antibodies 4 weeks after vaccination, the remainder were reimmunised and tested again after a further 4 weeks. Appropriate controls to test for tolerance induction and memory responses were included. The results indicate that fry are capable of mounting a humoral immune response very early in ontogeny. There is a period of 'unresponsiveness' which persists for longer against HGG, than against AS, though it was not thought to be tolerance as such. Memory could be detected to HGG in fry given a first immunisation at 2 months. The results are compared with preliminary experiments in which fry were first thymectomised 4 weeks before the first immunisation. In fry thymectomised at 1 month post hatch, and tested for primary and secondary responses at 2 and 3 months, the primary response to HGG is unaltered, but the secondary response is reduced. Both the primary and secondary response to AS is unaltered. When thymectomy is performed later, the effect on the secondary response to HGG is no longer apparent, but the primary response to AS is slightly reduced.     

Antibody response and duration of latent infection in sheep following experimental infection with Babesia ovis

Babesia ovis isolated in Extremadura (Spain) was the subject of a serological study in experimentally inoculated sheep. The first antibody titres, determined by the indirect fluorescent antibody (IFA) test, were observed 7-8 days post infection (p.i.) in all animals except the splenectomized group, in which the only animal that survived showed antibody response 10 days p.i. A faster response following challenge was observed in sheep which were seropositive before inoculation, which suggests the existence of an antigen memory. The highest titres were reached 16-25 days p.i., and subsequently began to fall, reaching minima at the end of the experimental period (330 days p.i.). The chronic carrier state in experimental B. ovis infection had a duration of at least 2 years. Passive transmission of antibodies from experimentally infected mothers to newborn lambs was also detected. Antibody levels were observed for a period not longer than 2 months after birth.     

The effect of the TLR9 ligand CpG-oligodeoxynucleotide on the protective immune response to alcelaphine herpesvirus-1-mediated malignant catarrhal fever in cattle

We wished to determine the effect of of CpG ODN adjuvant on the magnitude and duration of protective immunity against alcelaphine herpesvirus-1 (AlHV-1) malignant catarrhal fever (MCF), a fatal lymphoproliferative disease of cattle. Immunity was associated with a mucosal barrier of virus-neutralising antibody. The results showed that CpG ODN included either with emulsigen adjuvant and attenuated AlHV-1 (atAlHV-1) or alone with atAlHV-1 did not affect the overall protection from clinical disease or duration of immunity achieved using emulsigen and atAlHV-1. This is in contrast to other similar studies in cattle with BoHV-1 or cattle and pigs with various other immunogens. In addition to this, several other novel observations were made, not reported previously. Firstly, we were able to statistically verify that vaccine protection against MCF was associated with virus-neutralising antibodies (nAbs) in nasal secretions but was not associated with antibodies in blood plasma, nor with total virus-specific antibody (tAb) titres in either nasal secretions or blood plasma. Furthermore, CpG ODN alone as adjuvant did not support the generation of virus-neutralising antibodies. Secondly, there was a significant boost in tAb in animals with MCF comparing titres before and after challenge. This was not seen with protected animals. Finally, there was a strong IFN-γ response in animals with emulsigen and atAlHV-1 immunisation, as measured by IFN-γ secreting PBMC in culture (and a lack of IL-4) that was not affected by the inclusion of CpG ODN. This suggests that nAbs at the oro-nasal-pharyngeal region are important in protection against AlHV-1 MCF.     

Immunisation strategies against prion diseases: prime-boost immunisation with a PrP DNA vaccine containing foreign helper T-cell epitopes does not prevent mouse scrapie

Vaccination against prion diseases constitutes a promising approach for the treatment and prevention of the disease. Passive immunisation with antibodies binding to the cellular prion protein (PrP(C)) can protect against prion disease. However, immunotherapeutic strategies with active immunisation are limited due to the immune tolerance against the self-antigen. In order to develop an anti-prion vaccine, we designed a novel DNA fusion vaccine composed of mouse PrP and immune stimulatory helper T-cell epitopes of the tetanus toxin that have previously been reported to break tolerance to other self-antigens. This approach provoked a strong PrP(C)-specific humoral and cellular immune response in PrP null mice, but only low antibody titres were found in vaccinated wild-type mice. Furthermore, prime-boost immunisation with the DNA vaccine and recombinant PrP protein increased antibody titres in PrP null mice, but failed to protect wild-type mice from mouse scrapie.     

Development and duration of protection against salmonellosis in mice and sheep immunised with live aromatic-dependent Salmonella typhimurium.

The aim of this investigation was to determine the development and duration of protection in mice or sheep immunised with aromatic-dependent (aro-) Salmonella typhimurium strain CS332, by either parenteral or oral routes. Immunisation of mice by the intraperitoneal or sheep by the intramuscular routes was found to impart protection against oral challenge with the virulent parent S typhimurium strain CS94 as early as seven days after immunisation. In contrast, when immunisation was carried out by the oral route, protection was not evident until three weeks after immunisation. Regardless of the route of immunisation, mice were still partially protected at three months and were fully susceptible at six months after immunisation. In sheep, protection persisted for six months but not 12 months after immunisation. Only parenterally immunised mice and sheep developed high ELISA and, or, agglutinating antibody titres, and cutaneous delayed-type hypersensitivity (DTH) at three weeks after immunisation. Although both antibody and DTH were detectable three months after immunisation of mice with aro- S typhimurium strain CS332, none was detected at six months. Antibody measured by agglutination and ELISA was detectable six months after immunisation in sheep, although no DTH was evident. At 12 months after immunisation low levels of anti-LPS antibody (measurable by ELISA only) were detected in sheep immunised by the intramuscular route.     

Specific antibody-containing cells in the mammary gland of non-lactating sheep after intraperitoneal and intramammary immunisation

Ewes were immunised intraperitoneally with ovalbumin and Brucella abortus in Freund's complete adjuvant, followed seven days later by intramammary immunisation in which ovalbumin was presented to one mammary gland and Brucella abortus to the other. Mammary tissue taken after a further seven days contained more antigen-specific plasma cells than ewes given intraperitoneal or intramammary immunisation alone. These cells were found predominantly in the specifically immunised gland and only a few were found in the contralateral gland. Most of these cells were of the IgG1 isotype. There was also an increase in the total number of IgG1- and IgG2-containing cells in mammary gland tissues of these ewes, indicating a non-specific response to immunisation. Following either intraperitoneal or intramammary immunisation there was also a significant increase in the number of antigen-specific IgA cells in the lamina propria of the jejunum. The gut response following intramammary immunisation alone was abrogated by chronic drainage of intestinal lymph but not mammary lymph. This suggests that antigen may relocate from the mammary gland to the intestine where an IgA response is generated from gut associated lymphoid tissue. These data provide evidence for interaction between the gut and mammary gland of sheep in response to antigen.     

Involvement of spleen components of mature fowl in the primary and secondary humoral antibody response following experimental EDS'76 virus infection

Cellular changes in spleens of mature fowl in relation to both the primary and secondary humoral antibody response following experimental EDS'76 virus infection were studied. The influence of splenectomy on humoral antibody response was also examined. Experimental fowl had been naturally infected with fowl adenovirus (FAV) but did not possess precipitins to these viruses at the time of EDS'76 virus infection. Since EDS'76 infection provokes a recall of the group antibody to FAV, this infection simultaneously induces a primary response against EDS'76 virus and a secondary response due to the recall of the group antibody to FAV. HI and precipitating antibody to EDS'76 virus (primary response) were first detected at 6 and 8 days p.i. respectively. Curves of HI, precipitating and neutralising antibody titres were biphasic; the first peak (IgM peak) occurred at 10-11 days p.i., the second (IgG peak) at 16-28 days p.i. Precipitating antibodies to FAV (secondary response) were demonstrated from 4 days p.i. The curve of these antibody titres was also biphasic, with peaks at the same times as in the primary response. Based on HI and AGP testing of primary and secondary immune response in both splenectomised and non-splenectomised fowl it is concluded that in the primary response the spleen of the adult fowl is involved significantly in only IgM secretion, while in the secondary response it is likely that both IgM and IgG are secreted in considerable amounts. Clusters of lymphoblasts and plasmablasts were observed at 3 days p.i. in the red pulp. It is very likely that antigen-antibody complexes are formed from that time and circulate bound to the surface of lymphocytes. These antigen-loaded lymphocytes are 'picked up' from the blood stream by -red pulp macrophages, leading to enhanced formation of lymphoblasts in the red pulp. Great numbers of these cells (which are very probably IgM secreting cells) were present on days 6 and 7 p.i., but were no longer detectable after day 10 p.i. -macrophages of the macrophagal ellipsoidal corona (MEC), leading to significant enlargement of the periellipsoidal lymphoid tissue (PELT) by an increase of the number of lymphocytes observed from days 4-12 p.i. The MEC was significantly enlarged from 7-12 days p.i., very likely due to an increased number of macrophages. Following deposition of antigen in the white pulp, formation of follicles begins. The number of small, intact follicles including follicle precursors increased from 6 days p.i.(ABSTRACT TRUNCATED AT 400 WORDS)     

The antibody response to bovine lymphocyte alloantigens

Antibodies were raised against lymphocyte cell-surface antigens by multiple immunisations with purified lymphocytes or by the exchange of skin allografts. Eighteen of 21 cattle immunised with lymphocytes raised a detectable cytotoxic antibody response. The serum antibody from 10 responders recognized only common lymphocyte antigens, those antigens which are present on all peripheral blood lymphocytes. One animal responded only to B lymphocyte antigens while 7 others responded to both classes of antigens. The amount of antibody produced varied greatly between individuals; antibody titres ranged from 1 to 1028. Antibody raised early in the response was sensitive to treatment with 2-mercaptoethanol (2-ME) suggesting that IgM was the predominant class of immunoglobulin. Subsequently antibody became resistant to this treatment suggesting the appearance of IgG. The antibody responses following the exchange of skin grafts were very similar in all 12 cattle studied. High titred antibody to common lymphocyte antigens was detected in the serum 14 days after grafting. The early antibody activity was sensitive to 2-ME treatment but became totally resistant within 14 days. Total peak antibody titres ranged from 128-2048. Antibody to B lymphocyte antigens was identified in 8 of the 12 cattle. The responses to B lymphocyte antigens were similar to those against the more widely distributed common lymphocyte antigens with respect to time of antibody appearance, time of peak titre and sensitivity to 2-ME. Peak titres ranged from 2 to 32. The change in antibody specificity with time was also studied. Sera from 11 of the 18 cattle which had responded against lymphocytes showed an increase or broadening in reaction frequency as immunisations increased, suggesting the production of antibody to secondary specificities. In the cattle which had been skin grafted, the broadest reaction patterns were seen 14 to 21 days after grafting. The broadest reaction patterns were seen when the antibody responses were at their highest titre levels and narrowed as titres decreased.     

Titres of naturally occurring alloantibodies against feline blood group antigens in Turkish Van cats

Seventy-eight Turkish Van cats were examined for alloantibody titres, of which 42.3 per cent had type A blood and 57.7 per cent had type B blood. No type AB cats were found. All type B cats (n = 45) showed gross evidence of agglutinating anti-A antibody, with titres ranging from 2 to 256. Sixty-seven per cent of type B cats had anti-A antibody in their plasma, with titres ranging from 8 to 32. However, 13 per cent of type B cats had plasma alloantibody titres of less than 8 and 20 per cent had titres that were higher than 32. A total of 33 type A cats were also tested for anti-B alloantibody titres in their plasma. Among the type A plasma, gross agglutination at titres of 2 and greater than 2 were determined in 24 per cent and 36 per cent of samples, respectively. Microscopic agglutination was seen in an additional 18 per cent of plasma samples. There was no significant association between gender and plasma alloantibody titres of cats (P > 0.05).     

A Comparison of Methods for Measuring the Antibody Response in Mice and Cattle following Vaccination against Food and Mouth Disease

We present a comparison of methods for evaluating the potency of foot and mouth disease vaccine in the laboratory. The anti-FMDV antibodies (Ab) in vaccinated mice were tested by liquid phase (lp) ELISA, solid phase (sp) ELISA and virus neutralization (VN), and were compared with the Ab titres detected by lpELISA, which is the official test in Argentina for testing the potency of FMD vaccines and protection against a virulent challenge in cattle. The results demonstrated that it is possible to relate the Ab levels induced in vaccinated mice with both the Ab and protective responses elicited in cattle. Furthermore, it was found that the anti-FMDV Ab titres in mice detected by lpELISA 14 days after vaccination should be an accurate parameter for predicting the results of the challenge test in cattle. Thus, this test in mice appears to be an inexpensive and rapid alternative for testing FMD vaccines in cattle.     

The influence of reproductive physiology and nutrient supply on the periparturient relaxation of immunity to the gastrointestinal nematode Trichostrongylus colubriformis in Merino ewes

A pen experiment was conducted to investigate the interaction of early-weaning and nutrient supply on the periparturient relaxation of immunity to the gastrointestinal nematode (GIN) Trichostrongylus colubriformis in Merino ewes. Mixed-age pregnant and non-pregnant (dry) ewes were infected with 8,000 T. colubriformis L(3)/week, and fed either a high or low quality diet. Following parturition, lambs were either removed from their mothers at 2 days of age or allowed to continue suckling. Systemic immunity began to wane during late pregnancy with circulating eosinophils and plasma total antibody (Ab) levels declining from day -37 (relative to the midpoint of lambing) and day -24, respectively. Pregnant ewes fed the low quality diet exhibited an increasing faecal worm egg count (WEC) from day -24 and had higher intestinal worm burdens on day 13, whereas ewes fed the high quality diet had a delayed transient rise in WEC of lower magnitude. Dry and early-weaned ewes remained highly resistant to T. colubriformis at all times. In the post-lambing/lactation period, ewes fed the high quality diet had higher levels of local total Ab and numbers of goblet cells (GC) in the small intestine on days 13 and 41. Lactating/suckled ewes had a lower anti-parasite local immune response as indicated by reduced titres of total Ab, IgG(1), IgM and IgA and lower numbers of mucosal mast cells (MMC), globule leukocytes (GL) and GC in small intestinal tissue compared to their dry and early-weaned counterparts. Early-weaning resulted in rapid recovery of blood eosinophils and total Ab. On day 13 post-lambing, titres of total Ab, IgG(1), IgM, IgA and IgE, and numbers of MMC and GL were greater than those measured in dry and suckled ewes. When fed the high quality diet, ewes had a higher dry matter (DM) intake, maternal weight, fat score, greater fat depth and eye muscle depth, birthed heavier lambs that had higher growth rates, and produced more milk. The physiological status of pregnancy resulted in a higher DM intake but lower measures of fat depth and eye muscle depth, and suckling led to an increase in DM intake but a reduction in body weight and fat score through mobilisation of fat and muscle reserves. Despite the marked effect of diet quality on production traits, some inconsistencies were observed between body composition and apparent parasite resistance, measured by WEC and worm counts, suggesting that the nutritional influence was not necessarily always mediated through changes in body composition. Although reproductive status affected blood leptin levels, diet had no effect within suckled ewes and therefore it was concluded that leptin has no causative role in maintaining the periparturient relaxation of immunity to T. colubriformis.     

Involvement of spleen components of mature fowl in the primary and secondary humoral antibody response following experimental EDS'76 virus infection

Summary

Cellular changes in spleens of mature fowl in relation to both the primary and secondary humoral antibody response following experimentalEDS'76 virus infection were studied. The influence of splenectomy on humoral antibody response was also examined.

Experimental fowl had been naturally infected with fowl adenovirus (FAV) but did not possess precipitins to these viruses at the time of EDS’ 76 virus infection. Since EDS'76 infection provokes a recall of the group antibody to FAV, this infection simultaneously induces a primary response against EDS’ 76 virus and a secondary response due to the recall of the group antibody to FAV.

HI and precipitating antibodies toEDS'76 virus (primary response) werefirst detected at 6 and 8 days p.i. respectively. Curves of HI, precipating and neutralising antibody titres were biphasic; the first peak (IgM peak) occurred at 10–11 days p.i., the second (IgG peak) at 16–28 days p.i.

Precipitating antibodies to FAV (secondary response) were demonstrated from 4 days p.i. The curve of these antibody titres was also biphasic, with peaks at the same times as in the primary response.

Based on HI and AGP testing of primary and secondary immune response in both splenectomised and non‐splenectomised fowl it is concluded that in the primary response the spleen of the adult fowl is involved significantly in only IgM secretion, while in the secondary response it is likely that bothIgM and IgG are secreted in considerable amounts.

Clusters of lymphoblasts and plasmablasts were observed at 3 days p.i. in the red pulp. It is very likely that antigen‐antibody complexes are formed from that time and circulate bound to the surface of lymphocytes. These antigen‐loaded lymphocytes are ‘picked up’ from the blood stream by

– red pulp macrophages, leading to enhanced formation of lymphoblasts in the red pulp. Great numbers of these cells (which are very probably IgM secreting cells) were present on days 6 and 7 p.i., but were no longer detectable after day 10 p.i.

– macrophages of the macrophagalellipsoidal corona (MEC), leading to significant enlargement of the periellipsoidal lymphoid tissue(PELT) by an increase of the number of lymphocytes observedfrom days 4–12 p.i. The MEC was significantly enlarged from 7–12 days p.i., very likely due to an increased number of macrophages.

Following deposition of antigen in the white pulp, formation of follicles begins. The number of small, intact follicles includingfollicle precursors increasedfrom 6 days p.i. From 15 days p.i. to the end of the experiment both the number and size of follicles increased significantly.

Uptake and processing of antigen by macrophages is probably accompanied by death of some of these cells. This might explain the degenerative changes observed in large mesenchymal cells, probably macrophages, at 3 and 5 days p.i. in the red pulp and at 5 and 6 days especially in the MEC. Splenitis which was present at 3 and 5 days p.i. and oedema observed in and around ellipsoidal cells at 5 days p.i. may be due to mediators released from these degenerative macrophages.

A significantly increased number of follicles with lymphoblasts was seen from 2–15 days p.i. while lymphoblasts and plasmablasts were present in the PELT from 5–15 days p.i., but predominantly at 6 and 7 days p.i. It is likely that disruption of follicles and blast transformation of white pulp lymphoid cells are secondary response events. White pulp lymphoblastsand plasmablastsare probably IgG secreting cells.

Splenomegaly was observed at 3, 5 and 6 days after infection and was mainly due to swelling of red pulp macrophages and infiltration of granulocytes in the red pulp. Ellipsoidal and periellipsoidal changes could contribute to the splenomegaly at 5 and 6 days p.i.     

Kinetics and type of immune response following intranasal and subcutaneous immunisation of calves

Protection of animals against respiratory infections has long been known to depend on respiratory mucosal immunity. However, few studies have been reported on the immune response following intranasal (i.n.) immunisation with non-living, soluble antigens. This study determined the kinetics of the humoral and cellular immune responses in calves after i.n. immunisation with Limulus haemocyanin (LH) with cholera toxin adjuvant, or subcutaneous (s.c.) immunisation with LH in incomplete Freund's adjuvant. A proliferative response of peripheral blood mononuclear cells cultured in vitro with LH was observed in animals immunised 7-10 days after i.n. and s.c. immunisations with no significant differences between the two immunised groups. LH -specific antibody was present in the serum of animals immunised s.c. (IgM, IgG1 and IgG2) and i.n. (IgA). Although significant IgA responses were observed, i.n. immunisations in cattle with soluble protein antigens and cholera toxin as an adjuvant did not induce a strong systemic immune response.     

Passive transfer of maternal immunoglobulin isotype antibodies against tetanus and influenza and their effect on the response of foals to vaccination.

Influenza and tetanus-specific antibodies of the IgG sub-isotypes are posively transferred to foals via colostrum and inhibit their response to inactivated influenza vaccines and tetanus toxoid. High titres of influenza antibodies of IgGa and IgGb subisotypes and tetanus antibodies of the IgGa, IgGb and IgG(T) subisotypes were detected in postsucking serum samples collected from foals born to mares that had received booster doses of multicomponent vaccines during the last 2 months of gestation. Thereafter, titres declined in an exponential manner but were still detectable in all foals at age 26 weeks, regardless of whether they had been vaccinated prior to age 26 weeks. Mean +/- s.e. half-life of decline of influenza IgGa antibodies (27.0 +/- 2.3 days) was significantly shorter than that of influenza IgGb antibodies (39.1 +/- 2.7 days; P<0.005). Tetanus IgGa and IgGb antibodies declined with half-lives of 28.8 +/- 3.0 and 34.8 +/- 5.1 days, respectively. Titres of tetanus IgG(T) antibodies were substantially higher than those of influenza IgG(T) antibodies in postsucking samples and remained so through age 26 weeks, declining with a half-life of approximately 35 days. Postsucking titres of tetanus and influenza antibodies of the IgA isotype were low and declined rapidly to undetectable levels. Yearlings showed significant increases in titre of influenza IgGa, IgGb and IgG(T) subisotype antibodies but no increase in influenza IgA antibodies in response to 2 doses of multicomponent vaccines containing tetanus toxoid and inactivated influenza A-1 and A-2 antigens. Yearlings also showed strong tetanus IgGa, IgGb and IgG(T) subisotype responses to one dose of vaccine and a substantial further rise in titre in response to administration of a second dose 3 weeks later, but failed to show an increase in titre of tetanus IgA antibodies. The influenza and tetanus IgGa, IgGb and IgG(T) subisotype responses of 6-month-old foals to vaccination followed the same pattern as those shown by yearlings but titres were generally lower. In contrast, 3-month-old foals failed to show increases in titre of either influenza or tetanus IgG subisotypes in response to 2 doses of vaccine and generally needed 1-3 additional booster doses of vaccine to achieve titres similar to those achieved by yearlings after 2 doses. Based on the finding that maternal antibodies exert a significant inhibitory effect on the response of foals to tetanus toxoid and inactivated influenza antigens, it is recommended that primary immunisation of foals born to vaccinated mares should not commence before age 6 months.     

Immune response in farm animals infected or immunised with bacteria of Chlamydia sp. and Chlamydophila sp. genus

Bacteria from genera Chlamydia (Ch.) and Chlamydophila (Chl.) are very pathogenic and may infect humans and animals. They also may cause latent infection, especially in animals. In this paper we discuss the non-specific and specific cellular and humoral immunity in farm animals, after infection or immunisation with Chlamydia sp. and Chlamydophila sp. bacteria. It has been shown, that the infection or immunisation with the microorganisms influenced the activity of polimorphonuclear cells (PMN) and mononuclear cells (MN) in the process of phagocytosis. It has also been shown that the bacteria influenced the amount and activity of lysozyme, activities of myeloperoxidase and lysosomal enzymes. Infection or immunisation with the microorganisms was demonstrated to affect numbers of lymphocytes T and B and those of their subpopulation as well as the activity of cytokines and levels of serum and secreted immunoglobulins. The changes were detected just a few hours after infection or immunisation and persisted for a few days to a few decades.     

Effect of corticosterone infusion on plasma corticosterone concentration, antibody production, circulating leukocytes and growth in chicken lines selected for humoral immune responsiveness

The effect of corticosterone on antibody production was studied in chicken lines selected for humoral immune response. 2. Twelve cockerels (33 days old) from lines selected for high or low antibody responses after immunisation with sheep red blood cells (SRBC) were implanted with mini-infusion pumps delivering corticosterone or vehicle continuously for 14 d. 3. Three days after implantation, the chickens were immunised intramuscularly with 0.25 ml packed SRBC. Blood samples were taken before implantation, before immunisation and 3, 5, 7 and 11 d after immunization. 4. Corticosterone infusion induced higher plasma corticosterone concentrations and heterophil/lymphocyte ratios than infusion of vehicle only. Growth was considerably depressed and relative weights of the thymus, bursa of Fabricius and spleen were less in the corticosterone-infused chickens. 5. An effect of corticosterone on antibody production could not be demonstrated, and differences between selection lines were unaffected.     

Polyclonal activation of B cells occurs in lymphoid organs from porcine reproductive and respiratory syndrome virus (PRRSV)-infected pigs

Porcine reproductive and respiratory syndrome virus (PRRSV) induces a persistent viral infection associated with an inefficient humoral immune response. A study of lymphoid B cells and specific humoral immune response was performed in blood and several lymphoid organs collected from PRRSV experimentally-infected pigs. Groups of specific pathogen-free (SPF) pigs were infected with the LHVA-93-3 isolate of PRRSV, and blood, tonsils, spleen and mediastinal lymph nodes (MLN) were collected at various times postinfection (p.i.) (3-60 days). Lymphoid cells were isolated, immunolabeled for cytofluorometric determination of B cell percentages, used for counting specific anti-PRRSV antibody secreting B cells by an ELISPOT assay, or cultured for metabolic activity. The presence of anti-PRRSV antibodies in the serum of infected pigs was determined using a commercial ELISA assay. Virus detection was performed in all tissues, including lungs, by virus isolation and RT-PCR. The results show that percentages of B cells increased in tonsils as soon as 3 days until 17 days p.i. in PRRSV-infected pigs while they increased in spleen at 3 days p.i. only, due to an increase of larger Ig(high)-producing B cells. Metabolic activity of lymphoid cells from blood and spleen increased at 3 days p.i. only while lymphoid cells from tonsils and MLN transiently decreased at that time and increased thereafter up to 60 days p.i. Anti-PRRSV antibody-secreting B cells occurred in tonsils after 10 days p.i. and strongly increased up to 60 days p.i. However, specific anti-PRRSV-secreting B cells were detected in blood and spleen after 17 days p.i and in MLN only after 45 days p.i. Specific antibodies were detectable in serum at 10 days p.i., reached the maximum level at 45 days and remained high up to 60 days p.i. Infectious virus was detected in lungs and MLN as soon as 3 days p.i., and remained detectable up to 45 days p.i. in tonsils of one pig while viral RNA was detected in most organs up to 60 days p.i. In vitro experiments revealed that inactivated virus induced a stimulation of lymphoid cells isolated from PRRSV-infected pigs while it was cytotoxic for lymphoid cells from control pigs. Taken together, these results indicate that viral infection induced simultaneously a polyclonal activation of B cells, mainly in tonsils, and an exaggerated and prolonged specific humoral immune response due to persistent viral infection in lymphoid organs.