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Abstract: There are several indications that periparturient depression of functional properties of polymorphonuclear neutrophilic granulocytes (PMN) may be of great importance for the pathogenesis of genital infections after calving. As the periparturient period is characterized by high plasma levels of corticosteroids, the hypothesis to be tested was that high concentrations of glucocorticoids during the periparturient period suppress uterine PMN in number and functionality. An in vivo endometritis model was applied to examine uterine PMN. Recombinant human interleukin-8 (rhIL-8) was infused into the uterus of estrous cows and heifers 24h after pretreatment with dexamethasone (0.07mg/kg i.m.). Six hours after rhIL-8 infusion high numbers of uterine PMN were isolated and characterized as to their immunophenotype and function. Animals treated with dexamethasone animals showed leucocytosis due to neutrophilia in peripheral blood. Despite a downregulation of expressed l-selectin, cattle treated with dexamethasone showed more uterine PMN than those treated with placebo. Dexamethasone decreased plasma concentrations of the immunomodulatory steroidal hormones cortisol and estrogen. Dexamethasone directly reduced the generation of reactive oxygen species (ROS) by uterine PMN. This may be a useful mechanism since it would protect the endometrium from tissue damage by excessive extracellular ROS. However, it is not known if the net effect is in fact a reduction in ROS, as the number of uterine cells increases. Our study shows that glucocorticoids may not be considered immunosuppressive in all cases and may play an important role in the regulation of post partum uterine defense mechanisms. [Copyright &y& Elsevier]

It has been suggested that in cases of puerperal endometritis of cattle infected with Escherichia coli and Arcanobacterium pyogenes, the neutrophils are compromised in their defense capacity or downregulated functionally. In addition to direct bacterial effects, contents of lochial secretions and secreted products of locally activated polymorphonuclear neutrophilic granulocytes (PMNs) may also account for changes in function of freshly immigrating neutrophils. In this study, lochial secretions were obtained from healthy cows and from cows infected by E. coli or A. pyogenes. Separated uterine PMN of infected cows displayed an altered phenotype and function which correlated with the degree of bacterial contamination. Concurrently tested circulating PMN showed no such changes. Infected lochial secretions sterilized by filtration also changed the phenotype of blood PMN. Lochial secretions of healthy cows displayed only minor effects. The effects on PMN function in infected cows varied: ingestion was less affected, whereas generation of reactive oxygen species (ROS) was severely depressed. Concurrently tested purified bacterial products (solubles and fragments) of E. coli and A. pyogenes did not induce the phenotypical and functional changes observed in blood PMN. Since infected lochia also contained high numbers of immigrated and probably activated PMN, the influence of supernatants from phorbol myristate acetate-activated PMN were tested on freshly isolated blood PMN. Such supernatants also increased the expression of certain surface molecules and inhibited the ROS generation. Thus, reduced function and altered phenotypes of PMN which immigrate into the uteri of cows with bacterial endometritis is due not only to interactions with bacteria or bacterial products, but is also to the uterine milieu. [Copyright &y& Elsevier]

In order to investigate pathogenic mechanisms of acute endometritis in cows and mares, we established an in vivo model in both species. Based on the results of an in vitro transmigration system, human recombinant interleukin-8 (rhIL-8; 1.25 μg per mare and 5 μg per cow in 50 ml phosphate-buffered saline) was used to attract polymorphonuclear neutrophil granulocytes (PMNs) into the uteri. Peak numbers of uterine neutrophils were attracted after 6 h, in both cows and mares. On average, mares responded more sensitively than cows, with 15 times higher numbers of rhIL-8-attracted uterine neutrophils (72±8×107 cells). In contrast to in vitro studies, in vivo migrated neutrophils (uterine neutrophils) of both species displayed a significantly reduced MHC class I expression. Expression of the CD11a molecule was significantly enhanced on equine uterine neutrophils but downregulated on bovine cells. Compared with untreated autologous peripheral neutrophils, both uterine and in vitro migrated neutrophils showed no alteration of phagocytic capacity. The ability to generate reactive oxygen species (ROS) was significantly upregulated in bovine and equine uterine neutrophils. This was also observed after in vitro migration of equine neutrophils, whereas ROS generation by bovine neutrophils was significantly depressed. In summary, the concept of inducing endometritis directly by local application of human interleukin-8 has been reliably successful in cows and mares. The model permits the analysis of PMN migration into the uterus under defined and controlled conditions. The observed differences between cows and mares with respect to phenotypical and functional characteristics of in vivo attracted uterine cells point to species-related features of neutrophil migration. In vitro transmigrated bovine and equine cells partially differ in phenotype and function from uterine neutrophils. Therefore, the in vitro transmigration assay cannot completely represent the in vivo endometritis model described here. [Copyright &y& Elsevier]