J.B. Dixon

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 humoral immune responses to keyhole limpet haemocyanin (KLH). In a 2 x 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 IgG1 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 humoral 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.

Level of nutrition and age at weaning: effects on humoral immunity in young calves

Thirty-two calves were studied to determine the effects of level of nutrition (400 and 1000 g (air-dry matter) milk substitute per d) and age at weaning (5, 9 and 13 weeks) on humoral immune responses measured in serum and bronchoalveolar washings (BAW). All calves were immunized with Keyhole limpet haemocyanin (KLH) at 3 weeks of age, and with horse erythrocytes (HRBC) 1 d after weaning. Feeding the higher level of nutrition compared with the lower level decreased anti-HRBC titres and serum IgG2 and IgA responses to KLH (P < 0.05). Weaning at 5 compared with 9 weeks of age decreased serum anti-HRBC responses (P < 0.05), but weaning age had no effect on anti-KLH responses (P > 0.05). Feeding the higher level of nutrition increased total protein (P < 0.05) and IgG2 concentrations (P < 0.01) in BAW. The results showed that variation in husbandry conditions that is within conventional limits affects humoral immune responses in young, artificially-reared calves.

Immune recognition of Echinococcus granulosus. 1. Parasite-activated, primary transformation by normal murine lymph node cells

Summary Culture of murine lymph node cells together with living protoscolices of Echinoccus granulosus is described. The presence of the parasite induced potent blastic transformation in lymphocytes of unimmunized mice as indicated by tritiated thymidine incorporation. The response was markedly reduced by killing the parasite immediately prior to culture. No blastogenic activity was detectable in supernatants from living parasites cultured alone. Protoscolices from artificially infected syngeneic mice were effective stimuli, as were protoscolices from naturally infected horse and sheep. Stimulation was not detectably reduced by maintenance of the parasite in vitro for 72 h at 37°C or for 46 days at 4°C prior to culture. It is concluded that unprimed lymphocytes are stimulated to transform by surface contact with a stimulator synthesized, but not secreted, by the parasite. The biological significance of the reaction and its possible contribution to immunosuppression are discussed.

Interleukin-1-like activity in synovial fluids and sera of horses with arthritis.

Synovial fluid samples of horses with osteoarthritis were investigated to detect interleukin-1 (IL-1) activity which could contribute to the disease pathogenesis. Of the 32 samples tested, 12 (37.5 per cent) showed an augmented phytohaemagglutinin induced proliferation of C3H/HeJ mouse thymocytes. Positive results were also seen in horses with infected arthritis, osteochondritis, traumatic arthritis and undefined synovial effusions. Normal synovial fluid and sera from all groups failed to show any detectable IL-1 activity. Fractionation of synovial fluid showed that the IL-1 activity was in the 15 to 20 Kd fractions. In the absence of mitogen, synovial fluid failed to stimulate thymocytes and did not stimulate the growth of an interleukin-2 (IL-2) dependent CTLL cell line, but synovial fluid stimulated IL-2 release by mouse spleen cells incubated with suboptimal doses of lectin. Evidence of an IL-1 inhibitor in synovial fluid from osteoarthritic horses was provided by ultrafiltration experiments and by the inhibitory activity of synovial fluid at particular dilutions in the thymocyte assay. The presence of IL-1-like activity could be relevant in the pathogenesis of arthritis in horses.

Bovine immune recognition of Ostertagia ostertagi larval antigens.

Analysis of a detergent-solubilized somatic antigen of Ostertagia ostertagi 3rd stage larvae by SDS-PAGE and Western blotting has revealed two specific antigens with apparent molecular weights of 17 and 43 kD under reducing conditions. Probing of the Ostertagia ostertagi preparation with preinfection control sera has shown two cross-reacting antigens with apparent molecular weights of 67 and 81 kD. Both the 17 and the 43 kD antigens were recognised by IgG1 which was the predominant reactive subclass. FPLC fractionation of the crude extract with gel filtration and ion-exchange columns demonstrated immune reactivity in discrete peaks. Comparisons of ELISA and lymphocyte transformation showed antigenic components reactive with both antibodies and primed lymphocytes.

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 (five, 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.

A study of immunoregulation of BALB/c mice by Echinococcus granulosus equinus during prolonged infection

Two models of intraperitoneal infection with E. granulosus equinus by protoscolices and by cyst passage in BALB/c mice were used to provide mesenteric lymph node cells for adoptive cell transfer into syngeneic recipient normal responder mice. The cell transfer inocula were shown to have depleted Thy‐1 cells, but to be highly suppressive to the normal sheep erythrocyte response of the recipients. The nature of the depletion and non‐specific suppression, and the infectious nature of the latter, are discussed in relation to other examples of mitogenic stimulation resulting in non‐specific T cell suppressor activity. The functions of Ly‐2,3+ cells, not only as suppressor, but as alloreactive cytotoxic cells, are discussed as a possible, autoimmune explanation for the longevity of the parasite within the mouse model, in contradistinction to the predictable early rejection of analogous xenografts.

Echinococcus granulosus infection in mice: host responses during primary and secondary infection

The reaction of Balb/c mice to primary and secondary subcutaneous infection with Echinococcus granulosus protoscoleces (PSC) is described. From 3 to 14 days following primary exposure to PSC, draining lymph nodes increase in weight and there is expansion of T and B lymphocyte populations, enhancement of in vitro lymphocyte transformational responses and production of PSC-specific IgM and IgE antibodies. Despite the persistence of viable PSC in host tissues, lymphocyte responses decline to pre-infection values over the period 3-8 weeks post-infection. Secondary exposure to PSC immediately induces lymphoproliferation, enhancement of transformational responses, production of IgE antibody and encapsulation of PSC by inflammatory cells. Although specific antibody levels remain high until at least 8 weeks after challenge infection, lymphocyte activity begins to decline after 4 days and is profoundly suppressed by 10 days. Parasite viability appears to be significantly reduced in secondary, as opposed to primary, infection and is associated with the accumulation of large numbers of eosinophils, mast cells and macrophages in infected tissues.

Anaemia, diarrhoea and opportunistic infections in Fell ponies

This report summarises clinical and pathological observations on Fell pony foals with a range of signs that included ill thrift, anaemia, respiratory infection, glossal hyperkeratosis and diarrhoea. Some of the foals had normochromic, normocytic anaemia and some had low levels of plasma proteins, including immunoglobulin G. Antibiotic and supportive treatment was ineffective and all affected foals died or were killed on humane grounds. Postmortem examination of 12 foals and tissues from 2 other foals revealed a range of lesions that included glossal hyperkeratosis, typhlocolitis, intestinal cryptosporidiosis, granulomatous enteritis, proliferative and necrotising bronchiolitis consistent with adenovirus infection; lesions similar to those in the respiratory tract were present in the salivary gland and pancreas of individual foals. Lymphoid tissue was judged to be smaller than expected. These observations suggest the possibility of opportunistic infections secondary to some form of undefined immunocompromised state.

Effects of weaning on antibody responses in young calves

Two experiments were performed to investigate the effects of cessation of feeding milk substitute (weaning) on antibody responses to Keyhole Limpet haemocyanin (KLH) in young calves. In the first experiment, 47 calves were weaned at 42 days of age, while a further 47 calves remained unweaned until 84 days of age. All calves were immunized with KLH at 43 and 85 days of age. Serum IgG1, IgG2 and IgA responses to KLH were increased in the early weaned calves compared with the late weaned calves following both primary and secondary immunizations. In the second experiment, 24 calves were immunized with KLH at 21 days of age. Subsequently, at 35 days of age, eight calves were weaned, eight calves were sham-weaned and eight calves remained unweaned. Sham-weaning involved continuing to feed milk substitute of greatly decreased nutritional value in an attempt to isolate nutritional from behavioural effects of weaning. No effects of treatments on antibody responses were detected. It was concluded that weaning affects antibody responses to antigens given near weaning but not to antigens given 2 weeks before weaning.