In this study, we focused analyzing γδ T cells during bovine mammary gland inflammation induced by Streptococcus uberis. A mammary gland cell suspension was obtained using lavage 24, 48, 72, and 168 h after intramammary-induced infection. The proportion of lymphocytes increased during the entire week in which inflammation was present. The γδ T cells were also elevated during inflammation, reaching their peak at 72 h following induced inflammation. The percentage of apoptotic lymphocytes continually increased, with the highest proportion occurring 168 h after S. uberis infection. The results show that γδ T cells may be involved in the resolution of inflammation in bovine mammary glands, with the apoptosis of those cells potentially playing an important role.

AgroBioTech Research Centre Slovak University of Agriculture in Nitra 94976 Nitra Slovakia

Department of Animal Husbandry Sciences Faculty of Agriculture University of South Bohemia Studentska 1668 370 05 Ceske Budejovice Czech Republic

Department of Animal Morphology Physiology and Genetics Faculty of AgriSciences Mendel University in Brno Zemedelska 1 613 00 Brno Czech Republic

Department of Animal Nutrition and Forage Production Faculty of AgriSciences Mendel University in Brno Zemedelska 1 613 00 Brno Czech Republic

Department of Life Science and Bioinformatics Assam University Silchar 788 011 India

Faculty of Agrobiology and Food Resources Institute of Animal Husbandry Slovak Agriculture University in Nitra Trieda Andreja Hlinku 2 94976 Nitra Slovakia

Faculty of Biotechnology and Food Sciences Institute of Applied Biology Slovak University of Agriculture in Nitra Tr A Hlinku 2 94976 Nitra Slovakia

Infectious Diseases and Preventive Medicine Department Veterinary Research Institute Hudcova 296 70 621 00 Brno Czech Republic

NPPC Research Institute for Animal Production Hlohovecka 2 95141 Luzianky Slovakia

Zobrazit více v PubMed

Cobirka M., Tancin V., Slama P. Epidemiology and Classification of Mastitis. Animals. 2020;10:2212. doi: 10.3390/ani10122212. PubMed DOI PMC

Tamilselvam B., Almeida R.A., Dunlap J.R., Oliver S.P. Streptococcus uberis internalizes and persists in bovine mammary epithelial cells. Microb. Pathog. 2006;40:279–285. doi: 10.1016/j.micpath.2006.02.006. PubMed DOI

Almeida R.A., Luther D.A., Park H.M., Oliver S.P. Identification, isolation, and partial characterization of a novel Streptococcus uberis adhesion molecule (SUAM) Vet. Microbiol. 2006;115:183–191. doi: 10.1016/j.vetmic.2006.02.005. PubMed DOI

Almeida R.A., Dunlap J.R., Oliver S.P. Binding of host factors influences internalization and intracellular trafficking of Streptococcus uberis in bovine mammary epithelial cells. Vet. Med. Int. 2010 doi: 10.4061/2010/319192. PubMed DOI PMC

Almeida R.A., Kerro-Dego O., Headrick S.I., Lewis M.J., Oliver S.P. Role of Streptococcus uberis adhesion molecule in the pathogenesis of Streptococcus uberis mastitis. Vet. Microbiol. 2015;179:332–335. doi: 10.1016/j.vetmic.2015.07.005. PubMed DOI

Almeida R.A., Kerro-Dego O., Prado M.E., Headrick S.I., Lewis M.J., Siebert L.J., Pighetti G.M., Oliver S.P. Protective effect of anti-SUAM antibodies on Streptococcus uberis mastitis. Vet. Res. 2015;46:133. doi: 10.1186/s13567-015-0271-3. PubMed DOI PMC

Reinoso E.B. Bovine mastitis caused by Streptococcus uberis: Virulence factors and biofilm. J. Microb. Biochem. Technol. 2017;9:5.

Sladek Z., Rysanek D., Ryznarova H., Faldyna M. The role of neutrophil apoptosis during experimentally induced Streptococcus uberis mastitis. Vet. Med. Czech. 2006;51:437–447. doi: 10.17221/5571-VETMED. DOI

Slama P., Sladek Z., Rysanek D., Langrova T. Effect of Staphylococcus aureus and Streptococcus uberis on apoptosis of bovine mammary gland lymphocytes. Res. Vet. Sci. 2009;87:233–238. doi: 10.1016/j.rvsc.2009.03.005. PubMed DOI

Slama P., Sladek Z., Rysanek D. Lipopolysaccharide delays apoptosis of bovine lymphocytes. FEBS J. 2009;276:223.

Slama P., Zavadilova T., Kratochvilova L., Kharkevich K., Uhrincat M., Tancin V. Effect of peptidoglycan of Staphylococcus aureus on apoptosis of bovine mammary gland lymphocytes. J. Microbiol. Biotech. Food Sci. 2019;9:445–446. doi: 10.15414/jmbfs.2019.9.special.445-446. DOI

Slama P., Kabourkova E., Sladek Z., Zavadilova T., Kratochvilova L., Kharkevich K., Roychoudhury S., Pavlik A., Roztocilova A., Uhrincat M., et al. Effect of Lipopolysaccharide and Muramyl Dipeptide on Apoptosis of Bovine Mammary Gland Lymphocytes. Animals. 2020;10:990. doi: 10.3390/ani10060990. PubMed DOI PMC

Guzman E., Hope J., Taylor G., Smith A.L., Cubillos-Zapata C., Charleston B. Bovine γδ T cells are a major regulatory T cell subset. J. Immunol. 2014;193:208–222. doi: 10.4049/jimmunol.1303398. PubMed DOI PMC

Zhao Y., Niu C., Cui J. Gamma-delta (γδ) T cells: Friend or foe in cancer development? J. Transl. Med. 2018;16:3. doi: 10.1186/s12967-017-1378-2. PubMed DOI PMC

Sustrova T., Slama P. The effect of Staphylococcus aureus bacteria to proportion of gamma delta T-lymphocytes from bovine mammary gland. In: Skarpa P., Ryant P., Cerkal R., Polak O., Kovarnik J., editors. MENDELNET 2013, Proceedings of the 20th International PhD Students Conference MENDELNET 2013, Brno, Czech Republic, 20–21 November 2013. Mendel University in Brno; Brno, Czech Republic: 2013. pp. 788–792.

Slama P., Sladek Z., Kabourkova E., Havlicek Z., Kwak J.Y. Apoptosis of gamma delta T cells during inflammatory re-sponse of bovine mammary gland induced by Staphylococcus aureus. Eur. J. Immunol. 2016;46((Suppl. S1)):495.

Telfer J.C., Baldwin C.L. Bovine gamma delta T cells and the function of gamma delta T cell specific WC1 co-receptors. Cell Immunol. 2015;296:76–86. doi: 10.1016/j.cellimm.2015.05.003. PubMed DOI

Baldwin C.L., Telfer J.C. The bovine model for elucidating the role of γδ T cells in controlling infectious diseases of importance to cattle and humans. Mol. Immunol. 2015;66:35–47. doi: 10.1016/j.molimm.2014.10.024. PubMed DOI

Faldyna M., Leva L., Sladek Z., Rysanek D., Toman M. γδ-TCR+ CD2– lymphocytes are recruited into bovine mammary gland after stimulation. Vet. Med. Czech. 2006;51:258–264. doi: 10.17221/5545-VETMED. DOI

Sladek Z., Rysanek D., Faldyna M. Activation of phagocytes during initiation and resolution of mammary gland injury induced by lipopolysaccharide in heifers. Vet. Res. 2002;33:191–204. doi: 10.1051/vetres:2002007. PubMed DOI

Vermes I., Haanen C., Steffens-Nakken H., Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J. Immunol. Methods. 1995;184:39–51. doi: 10.1016/0022-1759(95)00072-I. PubMed DOI

Davis W.C., Brown W.C., Hamilton M.J., Wyatt C.R., Orden J.A., Khalid A.M., Naessens J. Analysis of monoclonal antibodies specific for the gamma delta TcR. Vet. Immunol. Immunopathol. 1996;52:275–283. doi: 10.1016/0165-2427(96)05578-X. PubMed DOI

Sladek Z., Rysanek D., Ryznarova H., Faldyna M. Neutrophil apoptosis during experimentally induced Staphylococcus aureus mastitis. Vet. Res. 2005;36:629–643. doi: 10.1051/vetres:2005023. PubMed DOI

Maxymiv N.G., Bharathan M., Mullarky I.K. Bovine mammary dendritic cells: A heterogeneous population, distinct from macrophages and similar in phenotype to afferent lymph veiled cells. Comp. Immunol. Microbiol. Infect. Dis. 2012;35:31–38. doi: 10.1016/j.cimid.2011.09.009. PubMed DOI

Ismaili J., Olislagers V., Poupot R., Fournie J.J., Goldman M. Human γδ T Cells Induce Dendritic Cell Maturation. Clin. Immunol. 2002;103:296–302. doi: 10.1006/clim.2002.5218. PubMed DOI

Tian L., Choi S.C., Lee H.N., Murakami Y., Qi C.F., Sengottuvelu M., Voss O., Krzewski K., Coligan J.E. Enhanced efferocytosis by dendritic cells underlies memory T-cell expansion and susceptibility to autoimmune disease in CD300f-deficient mice. Cell Death Differ. 2016;23:1086–1096. doi: 10.1038/cdd.2015.161. PubMed DOI PMC

He Y., Wu K., Hu Y., Sheng L., Tie R., Wang B., Huang H. γδ T Cell and Other Immune Cells Crosstalk in Cellular Immunity. J. Immunol. Res. 2014;2014:960252. doi: 10.1155/2014/960252. PubMed DOI PMC

Espinosa-Cueto P., Magallanes-Puebla A., Castellanos C., Mancilla R. Dendritic cells that phagocytose apoptotic macrophages loaded with mycobacterial antigens activate CD8 T cells via crosspresentation. PLoS ONE. 2017;12:e0182126. doi: 10.1371/journal.pone.0182126. PubMed DOI PMC

Bannerman D.D., Paape M.J., Chockalingam A. Staphylococcus aureus intramammary infection elicits increased production of transforming growth factor-α, β1, and β2. Vet. Immunol. Immunopathol. 2006;112:309–315. doi: 10.1016/j.vetimm.2006.03.018. PubMed DOI

Andreotti C.S., Pereyra E.A.L., Baravalle C., Renna M.S., Ortega H.H., Calvinho L.F., Dallard B.E. Staphylococcus aureus chronic intramammary infection modifies protein expression of transforming growth factor beta (TGF-β) subfamily components during active involution. Res. Vet. Sci. 2014;96:5–14. doi: 10.1016/j.rvsc.2013.11.002. PubMed DOI

Bannermann D.D., Paape M.J., Lee J.W., Zhao X., Hoper J.C., Rainard P. Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection. Clin. Diagn. Lab. Immunol. 2004;11:463–472. doi: 10.1128/CDLI.11.3.463-472.2004. PubMed DOI PMC

Collins C.C., Bashant K., Erikson C., Thwe P.M., Fortner K.A., Wang H., Morita C.T., Budd R.C. Necroptosis of Dendritic Cells Promotes Activation of γδ T Cells. J. Innate Immun. 2016;8:479–492. doi: 10.1159/000446498. PubMed DOI PMC

Pasparakis M., Vandenabeele P. Necroptosis and its role in inflammation. Nature. 2015;517:311–320. doi: 10.1038/nature14191. PubMed DOI