Effect of probiotics on the viability of porcine and human neutrophils in vitro

DOI:10.17221/72/2017-VETMEDCitation:Sustrova T, Ondrackova P, Leva L, Kolarova M, Kulich P, Sladek Z (2017): Effect of probiotics on the viability of porcine and human neutrophils in vitro. Veterinarni Medicina, 62: 637-646.
download PDF

Lactobacillus, Bifidobacterium and Enterococcus cultures are increasingly used as probiotics for humans and pigs. The aim of this study was to investigate if co-cultivation of porcine and human neutrophils with probiotics can lead to increased apoptosis in vitro. Ten adult Large white pigs and 10 healthy human donors were used in this study. Neutrophils were isolated by dextran sedimentation and cultivated with and without the lactic acid bacteria Bifidobacterium bifidum, Lactobacillus rhamnosus, and Enterococcus faecium for 2, 4, 24 and 48 h. Early and late apoptosis was measured using flow cytometry, and cell lysis was detected based on lactate dehydrogenase activity (LDH). A significant (< 0.05; < 0.01) increase in apoptotic neutrophils and LDH was observed at 24 h and 48 h in vitro. All probiotics exerted their greatest effects on the early apoptosis of porcine neutrophils, while the effects of L. rhamnosus were most pronounced on late apoptosis and those of B. bifidum on LDH release of human neutrophils. The increased neutrophil apoptosis caused by probiotic bacteria can be beneficial for more efficient efferocytosis and faster resolution of inflammation and tissue regeneration. In conclusion, we have demonstrated that the interaction of B. bifidum, L. rhamnosus, and E. faecium with human and porcine neutrophils leads to their apoptosis.

References:
Angulo Sandra, Morales Albert, Danese Silvio, Llacuna Laura, Masamunt Maria Carme, Pultz Nicole, Cifone Maria Grazia, De Simone Claudio, Delgado Salvadora, Vila Jordi, Panés Julián, Donskey Curtis, Fernández-Checa Jose C., Fiocchi Claudio, Sans Miquel, Dalmasso Guillaume (2011): Probiotic Sonicates Selectively Induce Mucosal Immune Cells Apoptosis through Ceramide Generation via Neutral Sphingomyelinase. PLoS ONE, 6, e16953- doi:10.1371/journal.pone.0016953
 
Arunachalam K, Gill HS, Chandra RK (2000): Enhancement of natural immune function by dietary consumption of Bifidobacterium lactis (HN019). European Journal of Clinical Nutrition, 54, 263-267 doi:10.1038/sj.ejcn.1600938
 
Bekkering Siroon (2013): Another look at the life of a neutrophil. World Journal of Hematology, 2, 44- doi:10.5315/wjh.v2.i2.44
 
Carol M. (2006): Modulation of apoptosis in intestinal lymphocytes by a probiotic bacteria in Crohn's disease. Journal of Leukocyte Biology, 79, 917-922 doi:10.1189/jlb.0405188
 
Chiu Yi-Han, Hsieh Yi-Jen, Liao Kuang-Wen, Peng Kou-Cheng (2010): Preferential promotion of apoptosis of monocytes by Lactobacillus casei rhamnosus soluble factors. Clinical Nutrition, 29, 131-140 doi:10.1016/j.clnu.2009.07.004
 
Clark J. A. (2005): Epidermal growth factor reduces intestinal apoptosis in an experimental model of necrotizing enterocolitis. AJP: Gastrointestinal and Liver Physiology, 288, G755-G762 doi:10.1152/ajpgi.00172.2004
 
Daniluk U, Alifier M, Kaczmarski M (2012): Probiotic-induced apoptosis and its potential relevance to mucosal inflammation of gastrointestinal tract. Advances in Medical Sciences, 57, 175-182 doi:10.2478/v10039-012-0025-7
 
DeLeo F. R. (2004): Modulation of phagocyte apoptosis by bacterial pathogens. Apoptosis, 9, 399-413 doi:10.1023/B:APPT.0000031448.64969.fa
 
Dolgushin II, Andreeva IS, Plekhanova EV (2008): Interaction of neutrophils and different bacterial agents. Zhurnal Mikrobiologii, Epidemiologii, i Immunobiologii 5, 103–105.
 
Donnet-Hughes A., Rochat F., Serrant P., Aeschlimann J.M., Schiffrin E.J. (1999): Modulation of Nonspecific Mechanisms of Defense by Lactic Acid Bacteria: Effective Dose. Journal of Dairy Science, 82, 863-869 doi:10.3168/jds.S0022-0302(99)75304-X
 
Dykstra Natalie S., Hyde Lucie, MacKenzie Alexander, Mack David R. (2011): Lactobacillus plantarum 299v Prevents Caspase-Dependent Apoptosis In Vitro. Probiotics and Antimicrobial Proteins, 3, 21-26 doi:10.1007/s12602-011-9066-7
 
Elyoussoufi Zineb (2013): Induction of oxidative stress and apoptosis in human neutrophils by p-phenylenediamine. Journal of Toxicology and Environmental Health Sciences, 5, 142-149 doi:10.5897/JTEHS2013.0274
 
Fairbairn L., Kapetanovic R., Sester D. P., Hume D. A. (2011): The mononuclear phagocyte system of the pig as a model for understanding human innate immunity and disease. Journal of Leukocyte Biology, 89, 855-871 doi:10.1189/jlb.1110607
 
Fang He, Elina Tuomola, Heikki Arvilommi, Seppo Salminen (2000): Modulation of humoral immune response through probiotic intake. FEMS Immunology & Medical Microbiology, 29, 47-52 doi:10.1111/j.1574-695X.2000.tb01504.x
 
Fournier B M, Parkos C A (2012): The role of neutrophils during intestinal inflammation. Mucosal Immunology, 5, 354-366 doi:10.1038/mi.2012.24
 
Galdeano C. M., Perdigon G. (2006): The Probiotic Bacterium Lactobacillus casei Induces Activation of the Gut Mucosal Immune System through Innate Immunity. Clinical and Vaccine Immunology, 13, 219-226 doi:10.1128/CVI.13.2.219-226.2006
 
Hebert MJ, Takano T, Holthofer H, Brady HR (1996): Sequential morphologic events during apoptosis of human neutrophils. Modulation by lipoxygenase-derived eicosanoids. Journal of Immunology 1, 3105–3115.
 
Jinzhou Zhang, Tao He, Wensheng Chen, Wen Wang, Jincheng Liu, Qin Cui, Hailong Zhu, Weiyong Liu, Dinghua Yi (2008): Cyclooxygenase-2 Suppresses Polymorphonuclear Neutrophil Apoptosis After Acute Lung Injury. The Journal of Trauma: Injury, Infection, and Critical Care, 64, 1055-1060 doi:10.1097/TA.0b013e318047c07c
 
Kelly K. J., Sandoval R. M., Dunn K. W., Molitoris B. A., Dagher P. C. (2003): A novel method to determine specificity and sensitivity of the TUNEL reaction in the quantitation of apoptosis. AJP: Cell Physiology, 284, C1309-C1318 doi:10.1152/ajpcell.00353.2002
 
Khailova L., Mount Patrick S. K., Arganbright K. M., Halpern M. D., Kinouchi T., Dvorak B. (2010): Bifidobacterium bifidum reduces apoptosis in the intestinal epithelium in necrotizing enterocolitis. AJP: Gastrointestinal and Liver Physiology, 299, G1118-G1127 doi:10.1152/ajpgi.00131.2010
 
Klein Günter, Pack Alexander, Bonaparte Christine, Reuter Gerhard (1998): Taxonomy and physiology of probiotic lactic acid bacteria. International Journal of Food Microbiology, 41, 103-125 doi:10.1016/S0168-1605(98)00049-X
 
Lundqvist-Gustafsson H, Norrman S, Nilsson J, Wilsson A (2001): Involvement of p38-mitogen-activated protein kinase in Staphylococcus aureus-induced neutrophil apoptosis. Journal of Leukocyte Biology 70, 642–648.
 
Macha M, Taras D, Vahjen W, Arini A, Simon O (2004): Specific enumeration of the probiotic strain Enterococcus Faecium NCIMB 10415 in the intestinal tract and in faeces of piglets and sows. Archives of Animal Nutrition, 58, 443-452 doi:10.1080/00039420400020058
 
Martin Constance J, Peters Kristen N, Behar Samuel M (2014): Macrophages clean up: efferocytosis and microbial control. Current Opinion in Microbiology, 17, 17-23 doi:10.1016/j.mib.2013.10.007
 
Matalova E, Spanova A, Kovaru F (2003): Apoptotic DNA alterations in pig leukocytes after phagocytosis of bacteria are linked to maturation of the immune system. Physiological Research 52, 235–242.
 
MITSUOKA Tomotari (2014): Development of Functional Foods. Bioscience of Microbiota, Food and Health, 33, 117-128 doi:10.12938/bmfh.33.117
 
Nami Y., Abdullah N., Haghshenas B., Radiah D., Rosli R., Yari Khosroushahi A. (2014): A newly isolated probiotic Enterococcus faecalis strain from vagina microbiota enhances apoptosis of human cancer cells. Journal of Applied Microbiology, 117, 498-508 doi:10.1111/jam.12531
 
Nami Yousef, Haghshenas Babak, Haghshenas Minoo, Abdullah Norhafizah, Yari Khosroushahi Ahmad (2015): The Prophylactic Effect of Probiotic Enterococcus lactis IW5 against Different Human Cancer Cells. Frontiers in Microbiology, 6, - doi:10.3389/fmicb.2015.01317
 
Otte Jan-Michel, Mahjurian-Namari Rudja, Brand Stephan, Werner Ilka, Schmidt Wolfgang, Schmitz Frank (2009): Probiotics Regulate the Expression of COX-2 in Intestinal Epithelial Cells. Nutrition and Cancer, 61, 103-113 doi:10.1080/01635580802372625
 
Payne Claire M., Glasser Lewis, Tischler Marc E., Wyckoff Deborah, Cromey Douglas, Fiederlein Roger, Bohnert Oliver (1994): Programmed cell death of the normal human neutrophil: An in vitro model of senescence. Microscopy Research and Technique, 28, 327-344 doi:10.1002/jemt.1070280408
 
Plaza-Diaz Julio (2014): Modulation of immunity and inflammatory gene expression in the gut, in inflammatory diseases of the gut and in the liver by probiotics. World Journal of Gastroenterology, 20, 15632- doi:10.3748/wjg.v20.i42.15632
 
Ramachandran Anup, Madesh Muniswamy, Balasubramanian Kunissery A (2000): Apoptosis in the intestinal epithelium: Its relevance in normal and pathophysiological conditions. Journal of Gastroenterology and Hepatology, 15, 109-120 doi:10.1046/j.1440-1746.2000.02059.x
 
Ren Y., Stuart L., Lindberg F. P., Rosenkranz A. R., Chen Y., Mayadas T. N., Savill J. (2001): Nonphlogistic Clearance of Late Apoptotic Neutrophils by Macrophages: Efficient Phagocytosis Independent of  2 Integrins. The Journal of Immunology, 166, 4743-4750 doi:10.4049/jimmunol.166.7.4743
 
Reuter G (2001): The Lactobacillus and Bifidobacterium microflora of the human intestine: composition and succession. Current Issues in Intestinal Microbiology 2, 43–53.
 
Sakatani A, Fujiya M, Ueno N, Kashima S, Sasajima J, Moriichi K, Ikuta K, Tanabe H, Kohgo Y (2016): Polyphosphate derived from Lactobacillus brevis inhibits colon cancer progression through induction of cell apoptosis. Anticancer Research 26, 591–598.
 
Song S., Oh S., Lim K. T. (2015): The proteins (12 and 15 kDa) isolated from heat-killed Lactobacillus plantarum L67 induces apoptosis in HT-29 cells. Cell Biochemistry and Function, 33, 89-96 doi:10.1002/cbf.3094
 
Sulowska Zofia, Majewska Ewa, Klink Magdalena, Banasik Malgorzata, Tchórzewski Henryk (2005): Flow Cytometric Evaluation of Human Neutrophil Apoptosis During Nitric Oxide Generation In Vitro: The Role of Exogenous Antioxidants. Mediators of Inflammation, 2005, 81-87 doi:10.1155/MI.2005.81
 
Teahon K, Smethurst P, Levi A J, Menzies I S, Bjarnason I (1992): Intestinal permeability in patients with Crohn's disease and their first degree relatives.. Gut, 33, 320-323 doi:10.1136/gut.33.3.320
 
Vermes István, Haanen Clemens, Steffens-Nakken Helga, Reutellingsperger Chris (1995): A novel assay for apoptosis Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. Journal of Immunological Methods, 184, 39-51 doi:10.1016/0022-1759(95)00072-I
 
Weber D A, Sumagin R, McCall I C, Leoni G, Neumann P A, Andargachew R, Brazil J C, Medina-Contreras O, Denning T L, Nusrat A, Parkos C A (2014): Neutrophil-derived JAML inhibits repair of intestinal epithelial injury during acute inflammation. Mucosal Immunology, 7, 1221-1232 doi:10.1038/mi.2014.12
 
William R, Watson G, Redmond PH, Wang JH, Condron C, Bouchier-Hayes D (1996): Neutrophils undergo apoptosis following ingestion of Escherichia coli. Journal of Immunology 156, 3986–3992.
 
YAMAMOTO A., TANIUCHI S., TSUJI S., KOBAYASHI M. HASUI & Y., KOBAYASHI Y. (2002): Role of reactive oxygen species in neutrophil apoptosis following ingestion of heat-killed Staphylococcus aureus. Clinical & Experimental Immunology, 129, 479-484 doi:10.1046/j.1365-2249.2002.01930.x
 
Yan Fang, Polk D. Brent (2002): Probiotic Bacterium Prevents Cytokine-induced Apoptosis in Intestinal Epithelial Cells. Journal of Biological Chemistry, 277, 50959-50965 doi:10.1074/jbc.M207050200
 
Yang S, Reid G, Challis JR, Kim SO, Gloor GB, Bocking AD (2015): Is there a role for probiotics in the prevention of preterm birth? Frontiers in Immunology 17, 62.
 
Zelnickova Petra, Kovaru Hana, Pesak Sava, Lojek Antonin, Matalova Eva, Ondracek Jaroslav, Kovaru Frantisek (2006): Postnatal functional maturation of blood phagocytes in pig. Veterinary Immunology and Immunopathology, 113, 383-391 doi:10.1016/j.vetimm.2006.06.008
 
download PDF

© 2018 Czech Academy of Agricultural Sciences