Cell Culture System for Studying Bovine Neutrophil Diapedesis

Smits, Elke; Cifrian, E.; Guidry, A.J.; Rainard, Pascal; Burvenich, Christian; Paape, Max

doi:10.3168/jds.s0022-0302(96)76492-5

Cited by 34 publications

(34 citation statements)

References 29 publications

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“…In a subsequent study [88], involving a greater diversity of cows, it appeared that a variability exists among animals, with generated C5a concentrations ranging from 0.6 ng/mL to 20 ng/mL in the milk from uninflamed, uninfected quarters. Concentrations of 25 and 50 ng/mL have been shown to induce the migration of bovine neutrophils through a cell culture system comprising a mammary epithelial cell monolayer [105], whereas 5 ng/mL were without an effect. These findings suggest that the amount of C5a which can be generated in the milk of certain cows could be of functional significance, whereas others would not be.…”

Section: Recruitment Of Phagocytesmentioning

confidence: 98%

“…At other epithelial surfaces such as the lungs of rats, the outcome of the exposure of alveolar epithelial cells to C5a is the release of proinflammatory cytokines and the enhanced expression of the C5a receptor, leading to an enhanced phlogistic response [97]. There is some indirect evidence that mammary epithelial cells react to C5a [76,105]. If the contribution of C5a to the triggering of mammary inflammation proved true, it would be justified to address the origin of the variability of C5 concentration in milk (acquired or of genetic origin) [88], because it could have consequences on the cow'susceptibility to mastitis.…”

Section: The Biological Significance Of the Complement In Milk And Armentioning

confidence: 99%

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The complement in milk and defense of the bovine mammary gland against infections

Rainard

2003

Vet. Res.

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-The mammary gland of dairy cows, which is prone to infection by various bacteria, mobilizes local and systemic immune defenses to cope with pathogens. The complement system plays an important part in the innate immunity against microorganisms through its bactericidal, opsonic, and phlogistic functions. The amount of the complement in the milk of healthy glands of dairy cows is low. Moreover, the classical pathway of activation is not functional because of a shortage in C1q. By contrast, the alternative pathway is active, deposits C3b and C3bi on bacteria, and generates amounts of C5a which are highly variable among cows. A slight inhibition of the bactericidal/hemolytic activities, of the deposition of C1q on bacteria, and of the phlogistic activity of C5a makes milk a rather anti-inflammatory fluid. The inhibitory activity does not involve C3b/ C3bi deposition on bacteria, nor the generation of C5a by the alternative pathway. When inflammation develops, the blood-derived complement components overcome the inhibitions and complement-dependent bactericidal, opsonic and phlogistic activities may be high in milk. Further research is necessary to evaluate the contribution of C5a to the recruitment of leukocytes in the mammary gland, and to specify the links between the complement system and the response of resident cells (leukocytes and mammary epithelial cells) to infection stimulus. This will help to define the contribution of the complement system to resistance against mastitis, and could help to differentiate animals more or less resistant to this frequent and costly disease.

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Section: Recruitment Of Phagocytesmentioning

confidence: 98%

Section: The Biological Significance Of the Complement In Milk And Armentioning

confidence: 99%

The complement in milk and defense of the bovine mammary gland against infections

Rainard

2003

Vet. Res.

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“…Epithelial cells of the lung have been shown to possess receptors for C5a [10], but the presence of C5a receptors on the apical surface of mammary epithelial cells remains to be documented. In vitro experiments with purified bovine C5a have shown that C5a (> 25 ng/mL) provokes the migration of neutrophils through a confluent monolayer of mammary epithelial cells [18]. In vivo infusion of C5a in the teat cistern of cows also induces a moderate recruitment of neutrophils [13].…”

Section: Discussionmentioning

confidence: 99%

“…Concentrations of C5a (around 50 ng/mL) can be found in the milk of quarters suffering from clinical mastitis [17]. This concentration of C5a has been shown to induce the migration of bovine PMN through a confluent monolayer of mammary epithelial cells [18]. Moreover, infusion of 65 ng of C5a into the teat cistern of cows provokes an influx of PMN [13].…”

Section: Introductionmentioning

confidence: 99%

Bovine milk fat globules do not inhibit C5a chemotactic activity

Rainard

2002

Vet. Res.

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-The C5a complement fragment is a potent inflammatory molecule but its contribution to the inflammatory response of the mammary gland remains uncertain. One of the unresolved questions is the possible interference of whole milk with C5a. In this study, the chemotactic activity of purified bovine C5a was tested in the presence of whole or skimmed milk. Milk from healthy glands acted as a chemoattractant, which could mask any inhibitory effect on C5a activity. To circumvent milk activity, washed milk fat globules were incubated with optimal (1 nM) or suboptimal (0.1 nM) concentrations of C5a, and the eventual chemoattractant activity was measured. There was no reduction in C5a-induced migration through a polycarbonate filter or shape-change of neutrophils. The concentrations of C5a determined by ELISA in the fluid phase were not reduced after incubation with the fat globules. It can be concluded that bovine milk fat globules do not trap or degrade C5a. Although these results do not explain the inhibitory effect of whole milk in the C5a-induced recruitment of neutrophils in milk, they suggest that milk does not inhibit the second major activity of C5a (apart from chemotaxis), i.e. the stimulation of phagocytic and bactericidal activities of neutrophils. C5a / chemotactism / milk / fat globuleRésumé -Les globules gras du lait de vache n'inhibent pas l'activité chimiotactique du C5a. Le fragment du complément C5a est un puissant agent inflammatoire mais sa contribution à la réponse inflammatoire de la glande mammaire reste incertaine. Une des questions non résolues est l'interfé-rence possible du lait entier avec le C5a. Dans cette étude, l'activité chimiotactique du C5a bovin purifié a été évaluée en présence de lait entier ou écrémé. Le lait de glandes saines s'est révélé être un chimioattractant, ce qui pourrait masquer un éventuel effet inhibiteur sur le C5a. En vue de contourner l'activité du lait, des globules gras lavés ont été incubés avec des concentrations optimales (1 nM) ou suboptimales (0,1 nM) de C5a, et l'activité résultante a été mesurée. La migration à travers une membrane filtrante de polycarbonate ou la déformation des polynucléaires neutrophiles n'ont pas été diminuées. Les concentrations de C5a, mesurées par ELISA, n'ont pas été réduites après incubation 413 Vet. Res. 33 (2002) (33) 2 47 42 77 79; e-mail: rainard@tours.inra.fr avec les globules gras. Il peut en être conclu que les globules gras du lait ne capturent pas ou ne dégra-dent pas le C5a. Bien que ces résultats n'offrent pas d'explication à l'effet inhibiteur du lait entier sur le recrutement C5a-dépendant des neutrophiles dans le lait, ils suggèrent que le lait n'inhiberait pas l'autre activité majeure du C5a (en plus du chimiotactisme), à savoir la stimulation des fonctions phagocytaires et bactéricides des neutrophiles. C5a / chimiotactisme / lait / globules gras

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“…Although, to our knowledge, there are no publications available, this two-compartment system could potentially be applied to mammary cells as well, studying transport or metabolism of blood borne substances across the epithelium. However, interesting applications for mammary cells of the two-compartment system have been described previously in studies of polymorphonuclear neutrophil migration (Smits et al, 1996 and and protein and plasminogen activator production in mammary epithelial cells after extracellular calcium treatment (Cheli et al, 2001).…”

Section: Cell-based Models Of the Gastrointestinal Tractmentioning

confidence: 99%

Cell-based models to test the effects of milk-derived bioactives

Purup

Nielsen²

2012

Animal

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The life science industries have a strong interest in screening for novel bioactives in complex mixtures like milk and dairy products. Food bioactives are not only important for public health in general, but also have potential therapeutic applications for the treatment of a number of diseases. To identify these novel bioactives, establishment of robust screening assays is essential. The use of in vitro cell-based models for screening and testing have the advantage that several concentrations of mixtures or specific compounds can be assayed at the same time in cells from specific tissues. Primary cell cultures from target organs or established cell lines can be used to identify the most sensitive cells. In addition, a large number of transfected cell lines with very specific sensitivities have been developed. Different endpoints inherent to basal or more sophisticated cellular functions can be investigated, such as cell viability, apoptosis, migration, intracellular signalling, regulation of gene expression, morphology and metabolic alterations. The gastrointestinal tract is an obvious target for bioactive molecules delivered through milk and dairy products, because it lies at the interface between dietary components in the lumen and the internal processes of the host. Identification of bioactive factors that affects proliferation or migration of epithelial cells may have potential applications in promoting gastrointestinal health in both humans and animals. The mammary gland is another target organ of considerable interest since it has been estimated that up to 50% of all newly diagnosed breast cancers may be related to dietary factors. A large number of gastrointestinal and mammary epithelial cell lines are commercially available, but in order to study some cellular functions, primary cultures of freshly isolated cells are often preferred, as established cell lines do not always express specialised properties in culture.

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Cell Culture System for Studying Bovine Neutrophil Diapedesis

Cited by 34 publications

References 29 publications

The complement in milk and defense of the bovine mammary gland against infections

The complement in milk and defense of the bovine mammary gland against infections

Bovine milk fat globules do not inhibit C5a chemotactic activity

Cell-based models to test the effects of milk-derived bioactives

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