transforming-growth-factor-alpha and Mastitis--Bovine

The objective of the current observational study was to determine the potential associations between cow factors, clinical mastitis (CM) etiology, and concentrations of select acute phase proteins and cytokines in milk from affected quarters of cows with CM. Cows with CM (n=197) were grouped based on systemic disease severity, milk culture result, parity, days in milk (DIM), previous CM occurrence, and season of the year when CM occurred. Concentrations of lipopolysaccharide-binding protein (LBP), haptoglobin (Hp), BSA, IFN-gamma, tumor necrosis factor-alpha (TNF-alpha), IL-1beta, IL-8, IL-10, IL-12, transforming growth factor (TGF)-alpha, and TGF-beta and activity of lactate dehydrogenase (LDH) were evaluated. Differences in the least squares means log(10) transformed concentrations of these proteins were compared using multiple linear regression mixed models. The milk concentrations of LBP, Hp, IL-1beta, IL-10, and IL-12, and activity of LDH in milk were higher in cows with moderate to severe versus mild systemic disease. The concentrations of Hp, BSA, IL-1beta, and IL-10 in milk were higher in cows with a gram-negative versus gram-positive milk culture result. Season of the year when CM occurred was associated with the concentration of all proteins evaluated except for IL-1beta and IL-12. Concentrations were higher in the winter versus summer except for Hp and TGF-beta, for which the opposite was true. Concentrations of LBP, IL-10, and IL-12, and LDH activity in milk were associated with DIM group. Except for LBP, these proteins were lower in cows with CM during the first 60 DIM versus those in mid or later lactation. Interferon-gamma, TNF-alpha, and IL-8 were undetectable in 67, 31, and 20% of samples, respectively. Detection of IFN-gamma and IL-8 was associated with season, and detection of TNF-alpha and IL-8 was associated with systemic disease severity. The current study provides the most comprehensive report of milk concentrations of innate immune response proteins in cows with naturally occurring CM and identifies factors that potentially influence those concentrations. Further investigation into the seasonal variation of cytokine production and its potential effect on the outcome of CM is warranted. Furthermore, the results of this study provide useful data for planning future studies examining the role of the innate immune response in CM.

Topics: Acute-Phase Proteins; Animals; Cattle; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-8; L-Lactate Dehydrogenase; Mastitis, Bovine; Milk; Transforming Growth Factor alpha; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

In contrast to other mastitis pathogens, the host response evoked during Staphylococcus aureus intramammary infection is marked by the absence of the induction of critical cytokines, including IL-8 and TNF-alpha, which have established roles in mediating host innate immunity. The elucidation of changes in the expression of other mediators with the potential to regulate mammary inflammatory responses to S. aureus remains lacking. Transforming growth factor (TGF)-alpha, TGF-beta1, and TGF-beta2 are cytokines that regulate mammary gland development. Because these cytokines also have a demonstrated role in mediating inflammation, the objective of the current study was to determine whether S. aureus intramammary infection influences their expression. Ten cows were challenged with S. aureus and milk samples collected. Increases in milk levels of TGF-alpha were evident within 32h of infection and persisted for 16h. Increases in TGF-beta1 and TGF-beta2 levels were detected within 40h of S. aureus infection and persisted through the end of the study. Thus, in contrast to IL-8 and TNF-alpha, S. aureus elicits host production of TGF-alpha, TGF-beta1, and TGF-beta2. This finding may suggest a role for these cytokines in mediating mammary gland host innate immune responses to S. aureus.

Topics: Animals; Cattle; Cell Count; Enzyme-Linked Immunosorbent Assay; Female; Mastitis, Bovine; Milk; Staphylococcal Infections; Staphylococcus aureus; Transforming Growth Factor alpha; Transforming Growth Factor beta

Among the gram-negative bacteria that cause mastitis, Escherichia coli are the most prevalent. The innate immune system provides initial protection against E. coli infection by detecting the presence of the foreign pathogens and by mounting an inflammatory response, the latter of which is mediated by cytokines such as IL-1beta, IL-8, and tumor necrosis factor (TNF)-alpha. Although changes in these cytokines during mastitis have been well-described, it is believed that other mediators moderate mammary gland inflammatory responses as well. The growth factors/cytokines transforming growth factor (TGF)-alpha, TGF-beta1, and TGF-beta2 are all expressed in the mammary gland and have been implicated in regulating mammary gland development. In other tissues, these growth factors/cytokines have been shown to moderate inflammation. The objective of the current study was to determine whether TGF-alpha, TGF-beta1, and TGF-beta2 milk concentrations were altered during the course of E. coli-induced mastitis. The contralateral quarters of 11 midlactating Holstein cows were challenged with either saline or 72 cfu of E. coli, and milk samples were collected. Basal milk levels of TGF-alpha, TGF-beta1, and TGF-beta2 were 98.81 +/- 22.69 pg/mL, 3.35 +/- 0.49 ng/mL, and 22.36 +/- 3.78 ng/mL, respectively. Analysis of whey samples derived from E. coli-infected quarters revealed an increase in milk levels of TGF-alpha within 16 h of challenge, and these increases persisted for an additional 56 h. Elevated TGF-beta1 and TGF-beta2 milk concentrations were detected in E. coli-infected quarters 32 h after challenge, and these elevations were sustained throughout the study. Because TGF-alpha, TGF-beta1, and TGF-beta2 have been implicated in mediating inflammatory processes, their induction during mastitis is consistent with a role for these molecules in mediating mammary gland host innate immune responses to infection.

Topics: Animals; Cattle; Escherichia coli Infections; Female; Lactation; Mastitis, Bovine; Milk; Time Factors; Transforming Growth Factor alpha; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta2

Multiparous Holstein cows were infected in two quarters by intramammary infusion with Streptococcus agalactiae and slaughtered approximately 36 h later. Mammary tissue was removed from the infected quarters, uninfected contralateral quarters, and from pair-slaughtered uninfected controls; the tissue was frozen in liquid nitrogen. The RNA was extracted, and Northern blot analysis was performed for a variety of growth factors, stress-induced genes, milk protein genes, and control genes. Infection increased levels of mRNA coding for heat shock proteins 89 alpha, 89 beta, 70, 60, and 27. Simultaneously, concentrations of alpha-lactalbumin and casein mRNA decreased; alpha-lactalbumin mRNA showed a greater decline. The mRNA for several growth factors, including acidic fibroblast growth factor, basic fibroblast growth factor, epidermal growth factor, transforming growth factor-alpha, IGF-I, and IGF-II, were also increased as was the apoptosis marker, testosterone-repressed prostate mucin-2. Concentrations of mRNA for controls, beta-actin, and glyceraldehyde-3-phosphate dehydrogenase were unaffected. These results indicate that mastitis induces changes in the levels of mRNA encoding for a variety of peptide growth factors. Such changes in growth factors could be important in a variety of processes that occur during infection, such as protection against injury or tissue repair and recovery processes.

Topics: Animals; Caseins; Cattle; Epidermal Growth Factor; Female; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Growth Substances; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Lactalbumin; Mammary Glands, Animal; Mastitis, Bovine; RNA, Messenger; Transforming Growth Factor alpha