Blagitz Maiara G, Souza Fernando N, Batista Camila F, Azevedo Luis Fernando F, Benites Nilson Roberti, Melville Priscilla Anne, Diniz Soraia A, Silva Marcos X, Haddad João Paulo A, Heinnemann Marcos Bryan, Cerqueira Mônica M O P, Della Libera Alice M M P
Departamento de Clínica Médica,Faculdade de Medicina Veterinária e Zootecnia,Universidade de São Paulo,Av. Prof Dr Orlando Marques de Paiva,87,Cidade Universitária,São Paulo 05508-270,Brazil.
Departamento de Medicina Veterinária Preventiva,Escola de Veterinária,Universidade Federal de Minas Gerais,Belo Horizonte,Minas Gerais 31270-010,Brazil.
J Dairy Res. 2015 Nov;82(4):460-9. doi: 10.1017/S0022029915000308. Epub 2015 Jun 29.
Streptococcus dysgalactiae is a bacterium that accounts for a notable proportion of both clinical and subclinical intramammary infections (IMIs). Thus, the present study explores the function of milk neutrophils and the lymphocyte profile in mammary glands naturally infected with Streptococcus dysgalactiae. Here, we used 32 culture-negative control quarters from eight clinically healthy dairy cows with low somatic cell counts and 13 S. dysgalactiae-infected quarters from six dairy cows. Using flow cytometry, we evaluated the percentage of milk monocytes/macrophages and neutrophils, expression of CD62L, CD11b and CD44 by milk neutrophils, the levels of intracellular reactive oxygen species (ROS) production and phagocytosis of Staphylococcus aureus by milk neutrophils, and neutrophil viability. Furthermore, the percentages of B cell (CD21(+)) and T lymphocyte subsets (CD3(+)/CD4(+)/CD8(-); CD3(+)/CD8(+)/CD4(-); and CD3(+)/CD8(-)/CD4(-)), and the expression of CD25 by T milk lymphocytes (CD3(+)) and T CD4(+) milk cells were also assessed by flow cytometry using monoclonal antibodies. The present study showed a higher SCC and percentage of milk neutrophils, and a decrease in the percentage of milk monocytes/macrophages from S. dysgalactiae-infected quarters when compared to uninfected ones. We also observed a higher expression of CD11b by milk neutrophils and a tendency toward a decrease in neutrophil apoptosis rate in S. dysgalactiae-infected quarters. In addition, the S. dysgalactiae-infected quarters had higher percentages of milk T cells (CD3(+)) and their subset CD3(+)CD8(+)CD4(-) cells. Overall, the present study provided new insights into S. dysgalactiae IMIs, including distinct lymphocyte profiles, and a tendency toward an inhibition of apoptosis in milk neutrophils.
停乳链球菌是一种在临床和亚临床乳房内感染(IMIs)中占显著比例的细菌。因此,本研究探讨了天然感染停乳链球菌的乳腺中乳中性粒细胞的功能和淋巴细胞谱。在这里,我们使用了来自8头临床健康、体细胞计数低的奶牛的32个培养阴性对照乳腺区,以及来自6头奶牛的13个感染停乳链球菌的乳腺区。我们使用流式细胞术评估了乳单核细胞/巨噬细胞和中性粒细胞的百分比、乳中性粒细胞CD62L、CD11b和CD44的表达、细胞内活性氧(ROS)产生水平以及乳中性粒细胞对金黄色葡萄球菌的吞噬作用,还有中性粒细胞活力。此外,还使用单克隆抗体通过流式细胞术评估了B细胞(CD21(+))和T淋巴细胞亚群(CD3(+)/CD4(+)/CD8(-);CD3(+)/CD8(+)/CD4(-);以及CD3(+)/CD8(-)/CD4(-))的百分比,以及T乳淋巴细胞(CD3(+))和T CD4(+)乳细胞CD25的表达。本研究表明,与未感染的乳腺区相比,感染停乳链球菌的乳腺区体细胞计数(SCC)和乳中性粒细胞百分比更高,而乳单核细胞/巨噬细胞百分比降低。我们还观察到感染停乳链球菌的乳腺区乳中性粒细胞CD11b表达更高,且中性粒细胞凋亡率有降低趋势。此外,感染停乳链球菌的乳腺区乳T细胞(CD3(+))及其亚群CD3(+)CD8(+)CD4(-)细胞的百分比更高。总体而言,本研究为停乳链球菌引起的IMIs提供了新的见解,包括不同的淋巴细胞谱,以及乳中性粒细胞凋亡受到抑制的趋势。