J.K. Dineen

The role of immunologically specific and non-specific components of resistance in cross-protection to intestinal nematodes.

Abstract Vaccination of 6–8-month-old Merino sheep with irradiated Trichostrongylus colubriformis larvae gave a high level of protection (81 %) against single-species challenge with normal infective larvae of the same species. The level of protection (34%) was substantially reduced against challenge with a closely related species ( T. vitrinus ) and no significant protection occurred against single-species challenge with a generically unrelated nematode (Nematodirus spathiger). These results suggest that antigen(s) which stimulate protective immunity are shared by the related Trichostrongylus species but not by N. spathiger . By contrast with the results obtained for single-species challenge, vaccination with irradiated T. colubriformis produced 98–100% protection against all 3 species in animals challenged simultaneously with infective larvae of the 3 species. Comparison of the levels of protection recorded following the 2 types of challenge indicate that although a specific antigenic trigger is required to provoke an appropriate response, the results obtained, particularly in the case of N. spathiger , suggest that the terminal effector mechanism is not immunologically specific. The implications of these conclusions are discussed in relation to theories of the mechanism of resistance to gastrointestinal nematodes and the potential efficacy of vaccination in the field.

The response of lambs to vaccination at weaning with irradiated Trichostrongylus colubriformis larvae: segregation into 'responders' and 'non-responders'.

Abstract Dineen J. K. , Gregg P. and Lascelles A. K. 1978. The response of lambs to vaccination at weaning with irradiated Trichostrongylus colubriformis larvae: segregation into ‘responders’ and ‘non-responders’. International Journal for Parasitology 8 : 59–63. Groups of colostrum fed and colostrum deprived lambs were vaccinated with irradiated Trichostrongylus colubriformis larvae at weaning (3 months) and challenged with normal infective larvae. As there was no significant difference between the worm counts of these groups after challenge, it was concluded that the failure of lambs, generally, to respond to vaccination as vigorously as mature sheep, was not due to ‘feed-back inhibition’ of the immune response by the action of maternal antibody. However the results showed that the lambs segregated into ‘responders’ and ‘non-responders’. The mean worm count of responders was 1560 whereas for non-responders this was 24,340. A responder was defined as an animal whose worm count was less than the lower limit of the 99 % confidence interval of unvaccinated controls. These results suggest that genetically-determined factors play an important role in the responsiveness of lambs to vaccination. Whereas counts of globule leucocytes in duodenal tissues were negatively correlated with worm counts (i.e. positively correlated with resistance), counts of both eosinophils and neutrophils were positively correlated with worm counts. These findings suggest that either globule leucocytes are involved in the resistance mechanism, or they are by-products of cellular events involved in resistance. On the other hand it is unlikely that either eosinophils or neutrophils are involved in the mechanism of resistance.

The effect of age on the response of sheep to vaccination with irradiated trichostrongylus colubriformis larvae

Abstract Vaccination with irradiated Trichostrongylus colubriformis larvae produced a high level of immunity, as judged by faecal egg counts and worm burdens following challenge with normal larvae, in nine of ten sheep aged 10 months. In lambs aged 3 months, vaccination was less effective. Some lambs developed partial immunity, but others did not respond. Serum levels of antibodies to T. colubriformis acetylcholinesterase reflected the extent of antigenic exposure rather than the degree of immunity acquired, and there was no evidence that the unresponsiveness of the lambs was due to a deficiency in antibody production. Unresponsiveness was not associated with the numbers of circulating lymphocytes, monocytes or granulocytes, or with the numbers of mast cells, eosinophils and neutrophils at the site of infection. However, there were many globule leucocytes in the intestinal mucosa of adult sheep which were resistant to challenge infection. On the other hand, few of these cells were found in vaccinated lambs which generally gave a poor response to challenge.

An association between a lymphocyte antigen in sheep and the response to vaccination against the parasite Trichostrongylus colubriformis

Abstract Lymphocyte antigens were tested in sheep which had been selected for responsiveness to vaccination against the intestinal nematode Trichostrongylus colubriformis . These sheep had been bred in an assortative mating programme which produced offspring designated as either “high responders” or “low responders”, with highly heritable resistance or susceptibility. Ovine lymphocyte antigen (OLA) typing antisera were obtained from parous ewes in the course of matings which produced the high and low responder flocks. A particular antigen (SY1) was found to be present in high frequency on the lymphocytes of high responder (72·2%) and in lower frequency (21·9%) on the lymphocytes of low responder rams. In ewes, the frequency for high responders was 65·7% and for low responders it was 33·5%. A similar association between the SY1 antigen and low faecal egg count was found in random-bred sheep which had been vaccinated with irradiated larvae and challenged with normal larvae. The conclusion was drawn that this lymphocyte antigen was likely to be part of the sheep major histocompatibility complex which influenced the immune response of sheep to vaccination against the parasite.

The effect of sire selection on the response of lambs to vaccination with irradiated Trichostrongylus colubriformis larvae.

Abstract Dineen, J. K. and Windon, R. G. (1980). The effect of sire selection on the response of lambs to vaccination with irradiated Trichostrongylus colubriformis larvae. International Journal for Parasitology 10 : 189–196. Rams selected for responsiveness and unresponsiveness to vaccination with irradiated T. colubriformis larvae at an early age were mated to unselected random bred ewes. Progeny were vaccinated with 20,000 irradiated larvae at 8 and 12 weeks of age, given anthelmintic treatment at 16 weeks and challenged with 20,000 normai larvae at 17 weeks. The results, based on wether worm counts and ewe faecal egg counts, showed significant differences between responder and non-responder progeny. There was a significant correlation between worm counts and faecal egg counts of half-sibs from the same sire group. The occurrence of globule leucocytes was inversely related to worm burdens of wether progeny, however, no clear relationship was found with eosinophils. In vitro lymphocyte stimulation using T. colubriformis L 3 antigen, concanavalin A and lipopolysaccharide showed that statistically defined responder progeny, pooled from both responder and non-responder sire groups, gave higher responses than non-responder lambs after vaccination. The results confirm that genetically-determined factors are involved in the response of lambs to vaccination at an early age, and indicate that rapid genetic progress may be achieved in the type of mating usually carried out under fleld conditions.

The segregation of lambs into 'responders' and 'non-responders': response to vaccination with irradiated Trichostrongylus colubriformis larvae before weaning.

Abstract Random bred Merino ram and ewe lambs were vaccinated at 1, 2 and/or 3 months of age with irradiated T. colubriformis larvae. An exponentially increasing challenge of normal larvae was given to all groups including unvaccinated controls commencing at 1 month of age. The results, based on faecal egg counts, showed a dissociation into animals which responded to vaccination (geometric mean egg count 441) and those which did not (geometric mean egg count 1567). The proportion of responders was greatest in groups first vaccinated at the earliest age (1 month). Wool growth and liveweight gains showed severe depression corresponding to peak egg counts, however, responders were less affected than non-responders. There was no correlation between haemoglobin type and resistance to challenge. Faecal egg counts after impulse challenge with 10,000 normal larvae given at about 6 1 2 months of age showed a significant ranked correlation with those obtained during the primary exponential challenge. These results confirm that a proportion of young lambs respond to vaccination with irradiated larvae, and that genetically-determined factors are implicated in the ability of animals to respond to vaccination at an early age.

The effect of selection of both sire and dam on the response of F1 generation lambs to vaccination with irradiated Trichostrongylus colubriformis larvae

Abstract Windon R. G. and Dineen J. K. (1981). The effect of selection of both sire and dam on the response of F 1 generation lambs to vaccination with irradiated Trichostrongylus colubriformis larvae. International Journal for Parasitology 11 : 11–18. Rams and ewes, tested for responsiveness to vaccination with irradiated T. colubriformis larvae at an early age, were mated on the basis of responder × responder and non-responder × non-responder. Progeny were vaccinated at 8 and 12 weeks of age with 20,000 irradiated larvae, treated with anthelmintic at 16 weeks and challenged with 20,000 normal larvae at 17 weeks. Faecal egg counts of progeny from responder matings were significantly lower than progeny from non-responders, and within each mating type, ewe lambs had markedly lower egg counts than ram lambs. The level of circulating complement-fixing antibodies to T. colubriformis larval extract were inversely related to egg counts. Thus, ewe progeny from responder matings had the highest serum antibody levels, non-responder ram progeny had the lowest levels and responder rams and non-responder ewes had similar intermediate levels. In vitro responses of cells stimulated with T. colubriformis L 3 antigen were greater in progeny from responder matings, whereas responses to bacterial lipopolysaccharide were higher in progeny from non-responder matings. The results confirm that the response to vaccination at an early age is genetically determined, and show that the response of progeny is most vigorously expressed when both sires and dams have been selected.

The response of lambs to vaccination and challenge with Trichostrongylus colubriformis: Effect of plane of nutrition on, and the inter-relationship between, immunological responsiveness and resistance

Abstract Wagland B. M. , Steel J. W. , Windon R. G. and Dineen J. K. 1984. The response of lambs to vaccination and challenge with Trichostrongylus colubriformis : effect of plane of nutrition on, and the inter-relationship between, immunological responsiveness and resistance. International Journal for Parasitology 14: 39–44. Merino lambs weaned at 8 weeks of age were fed either ground and pelleted (high plane, HP) or chopped (low plane, LP) lucerne hay ad libitum to achieve an approximate 2-fold difference in liveweight gain. When aged 17 and 21 weeks, 15 of the 20 lambs in each diet group were vaccinated with 80,000 irradiated T. colubriformis larvae. At 25 weeks, vaccinated and unvaccinated lambs were treated with anthelmintic and one week later challenged with 30,000 normal larvae. Four weeks after challenge the animals were killed for worm counts. After vaccination HP lambs had higher titres of antibodies to the parasite and after challenge had lower worm egg outputs, and lower worm burdens than LP lambs. Immunological responsiveness (serum titre of complement-fixing antibodies against worm antigen) and manifestations of resistance (eggs produced per female worm per day and percent protection calculated from worm counts) were significantly correlated within dietary groups. Percent protection and egg production per female worm were highly correlated ( r = −0.81) in individual animals pooled over dietary groups, suggesting that both manifestations of resistance respond to essentially the same immunological mechanism. Failure to obtain significant correlation between weight gain pre-vaccination and immunological and resistance parameters indicated that animal production and resistance to infection are not genetically linked. Negative correlation of weight gain during the vaccination period with serum antibody titre at challenge suggests that the developing immune response competes with weight gain for limited physiological resources of the animal.

A novel host-protective antigen from Trichostrongylus colubriformis

The detergent-soluble fraction from Trichostrongylus colubriformis third-stage larvae contained a simple set of antigens, one of which (molecular weight 41,000) induced 43-51% protection in guinea pigs following immunization. Isolation and partial amino acid sequence analysis of this protective antigen showed it was parasite tropomyosin.

Nippostrongylus brasiliensis: effects of immunity on the pre-intestinal and intestinal larval stages of the parasite

Abstract Nippostrongylus brasiliensis : effects of immunity on the pre-intestinal and intestinal larval stages of the parasite. International journal for Parasitology 4: 183–191. Migration of the pre-intestinal larval stages of N. brasiliensis was studied in rats undergoing either primary or challenge infections. In rats undergoing a primary infection, more than 67 percent of larvae successfully migrated from the skin to the oesophagus by 70 h after infection, and subsequently over 90 per cent of these larvae became established in the small intestine as sexually mature adults. In immune rats undergoing a second infection, 46 per cent of larvae completed migration to the oesophagus by 70 h and of these, only 1·6 per cent became established in the intestine to produce eggs. These inhibitory effects on the pre-intestinal and intestinal larval stages were even more pronounced in immune rats undergoing a third or fourth infection and in addition, there was a prolonged sojourn and substantial retention of larvae in their lungs. There was no evidence that the immune response had an adverse effect on oesophageal fourth stags larvae as these organisms (obtained from immune donors) were able to establish and develop to maturity when transferred per os to normal animals. Syngeneic transfer of immune mesenteric lymph node cells to normal recipients, caused expulsion of parasites from the intestine but failed to effect migration of pre-intestinal larval stages. The implications of these findings are discussed in the context of current knowledge of the mechanisms of immunity to helminths.