Development of a method for isolating bovine colostrum mononuclear leukocytes for phenotyping and functional studies

Meganck, Vanessa; Goddeeris, Bruno; Stuyven, Edith; Piepers, Sofie; Cox, Eric; Opsomer, G.

doi:10.1016/j.tvjl.2014.02.021

Cited by 11 publications

(10 citation statements)

References 30 publications

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“…Apesar de os fagócitos mamários terem origem sanguínea, estudos demonstraram que eles podem ter sua capacidade fagocítica comprometida, em relação àqueles presentes na circulação sanguínea, por conta da fagocitose da gordura e caseína (PAAPE et al, 2003) e da curta meia-vida, levando à redução do burst respiratório e morte por apoptose (MEHRZAD et al, 2001), entretanto pouco se sabe ainda sobre a atividade funcional dessas células na secreção colostral, uma vez que a obtenção celular é prejudicada pela presença abundante de debris, gordura, proteínas e partículas autofluorescentes (GOMES et al, 2014;MEGANCK et al, 2014), além da baixa viabilidade (GODDEN et al, 2012).…”

Section: Palavras-chaveunclassified

“…Esse mecanismo compensatório foi também apresentado em outras investigações, como a identificação de maior capacidade proliferativa dos fagócitos mononucleares do colostro quando comparado ao sangue, provavelmente em virtude dos efeitos ativadores do colostro no fenótipo dos leucócitos (CONCHA et al, 1986;REBER et al, 2006), e que a maior quantidade de macrófagos no colostro poderia, em parte, balancear a menor resposta aos mitógenos e antígenos, quando comparados aos fagócitos sanguíneos (MEGANCK et al, 2014).…”

Section: Palavras-chaveunclassified

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Avaliação funcional dos fagócitos do sangue e da secreção mamária de vacas no período pós-parto imediato

et al. 2017

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RESUMO: O periparto caracteriza-se por mudanças imunes e fisiológicas na glândula mamária das fêmeas bovinas, tornando-as mais susceptiveis às mastites. Este trabalho teve o objetivo de avaliar a fagocitose da bactéria Escherichia coli pelos fagócitos do sangue e do colostro de vacas hígidas da raça Holandesa. Amostras provenientes de 22 fêmeas, entre novilhas e vacas, foram obtidas durante a primeira ordenha pós-parto, para avaliação da fagocitose, em lamínula de vidro, após estímulo com E. coli. A proporção de fagócitos espraiados do sangue foi maior quando as células foram estimuladas com bactérias (p=0,003), no entanto a taxa de fagocitose foi baixa e semelhante à observada quando as células não foram estimuladas (p=0,299). No colostro, foram constatados maiores índices de fagocitose (p=0,001) e espraiamento (p=0,000) quando os fagócitos foram estimulados com E. coli. Comparando-se os resultados obtidos para o sangue e o colostro, verificou-se, no sangue, maior proporção de fagócitos não espraiados e que não apresentavam bactérias no seu interior, estimulados (p=0,000) ou não (p=0,002) com bactéria. Houve maior taxa de espraiamento nos fagócitos do colostro quando estimulados com E. coli (p=0,002). Não se viu diferença nos percentuais de fagocitose entre as amostras de sangue e colostro incubadas (p=0,478) ou não (p=0,071) com bactérias. Os resultados permitiram concluir que E. coli influenciou no espraiamento dos fagócitos do sangue e do colostro de fêmeas bovinas da raça Holandesa no período do pós-parto imediato e auxiliarão na compreensão dos mecanismos imunológicos ocorridos nesse período, considerado o de maior susceptibilidade das fêmeas bovinas às infecções.

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Section: Palavras-chaveunclassified

Avaliação funcional dos fagócitos do sangue e da secreção mamária de vacas no período pós-parto imediato

et al. 2017

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“…Isolation of CMC was performed as described in Meganck et al (2014). Briefly, diluted colostrum samples were centrifuged (600 × g, 15 min, 18°C), the cream layer was removed, and cell pellets were washed twice.…”

Section: Isolation Of Cmcmentioning

confidence: 99%

“…The flow cytometric analysis was performed as described in Meganck et al (2014). Monoclonal antibodies used were mouse anti-bovine cluster of differentiation (CD) 21 (MCA1424G, AbDSerotec, Puchheim, Germany) for B-lymphocytes; mouse anti-bovine CD2 (BAQ95A, VMRD, Pullman, WA), expressed on T-lymphocytes and NK-cells; and mouse anti-bovine CD14 (MM61A, VMRD) for monocytes or macrophages.…”

Section: Leukocyte Phenotypic Analysis By Flow Cytometrymentioning

confidence: 99%

Effect of β-hydroxybutyric acid, parity, and body condition score on phenotype and proliferative capacity of colostral mononuclear leukocytes of high-yielding dairy cows

Meganck

Goddeeris

Campeneere³

et al. 2015

Journal of Dairy Science

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In neonatal calves, the ingestion of colostrum is imperative for preventing infectious diseases. Investigations into the transfer of passive immunity of cattle have primarily focused on the importance of colostral immunoglobulins, with a recent increase in focus on understanding the role of colostral leukocytes. The main objective of the present study was to measure the influence of parity, body condition score, serum nonesterified fatty acids, and serum β-hydroxybutyrate concentrations of periparturient cows on phenotype and mitogen-and antigen-induced proliferative capacity of bovine colostral leukocytes. Holstein-Friesian cows (n = 141) were intramuscularly vaccinated at 60 and 30 d before the expected parturition date with a tetanus toxoid vaccine. Of these 141 animals, 28 primiparous and 72 multiparous cows were sampled immediately after parturition. Colostrum mononuclear cell populations were identified by flow cytometry using bovine cluster of differentiation markers, and the proliferative capacity of these cells was determined using a 3 H-thymidine proliferation assay. Under-conditioned cows had a significantly higher percentage of colostral macrophages than normal-conditioned animals, whereas over-conditioned cows had significantly more colostral B-lymphocytes. Serum β-hydroxybutyrate was significantly associated with higher numbers of colostral T-lymphocytes and macrophages. Heifers had significantly higher mitogen-and antigen-induced proliferation of their colostral leukocytes than third parity or older cows. In conclusion, body condition score, parity, and serum β-hydroxybutyrate concentration of periparturient high-yielding dairy cows were shown to influence the number of colostral macrophages or the mitogen-and antigen-induced proliferation of colostral leukocytes, possibly influencing the cellular immunity of the newborn calf.

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“…Study of the peculiarities of the innate immune response in the immediate postpartum period is limited by the difficulty of isolating colostrum mononuclear leukocytes (Meganck et al 2014). Thus, the knowledge available comes from studies that were carried out with phagocytes obtained from blood or milk.…”

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Release of hydrogen peroxide by phagocytes from bovine colostrum in the peripartum period

et al. 2014

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Changes in the composition of colostrum in the peripartum period focused on the neonate's immunity can minimize the response of the mammary gland before natural exposure to mastitis pathogens. This research evaluated the oxidative activity of mononuclear and polymorphonuclear phagocytes of bovine colostrum. Bacteriologically negative (n = 171) bovine colostrum samples of first and second milking postpartum were evaluated. The oxidative activity was assessed by the production of hydrogen peroxide (H 2 O 2 ) released either spontaneously or upon stimulation by enterotoxigenic Escherichia coli (ETEC). For this, three treatments were used: control (C) -only cells; non-opsonized (NO) -cells and ETEC non-opsonized; and opsonized (O) -cells and ETEC opsonized. Median values of concentrations (nmol/10 6 ) of H 2 O 2 produced by cells obtained from the first and second milking treatments C, NO and O were 149.1 and 39.9 (P = 0.0007); 125.5 and 49.1 (P = 0.0007); 102.4 and 54.4 (P = 0.008), respectively. No differences were observed between the treatments at each milking. The amount of H 2 O 2 produced at first milking exceeded the values found thereafter. Furthermore, the presence of bacteria did not determine the increased amount of H 2 O 2 released by phagocytes. Thus, we concluded that the mammary gland's events during the peripartum period have an effect on the proportion and activity of phagocytes, which can cause injuries to the breast parenchyma by the large amount of free radicals produced. The high frequency of bovine mastitis during this period indicates the need for studies of the immunity of the mammary gland and research of susceptibility factors for bacterial infections.

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Development of a method for isolating bovine colostrum mononuclear leukocytes for phenotyping and functional studies

Cited by 11 publications

References 30 publications

Avaliação funcional dos fagócitos do sangue e da secreção mamária de vacas no período pós-parto imediato

Avaliação funcional dos fagócitos do sangue e da secreção mamária de vacas no período pós-parto imediato

Effect of β-hydroxybutyric acid, parity, and body condition score on phenotype and proliferative capacity of colostral mononuclear leukocytes of high-yielding dairy cows

Release of hydrogen peroxide by phagocytes from bovine colostrum in the peripartum period

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