Mastitis Süresince Meme Epitel Hücrelerinin Fonksiyonel Durumunu Değerlendirmek Amacıyla Sığır α-kazein Proteinine karşı Monoklonal Antikor Geliştirilmesi

Mastitis sütçü hayvanlarda yaygın bir hastalıktır ve dünya çapında büyük ekonomik kayıplara neden olmaktadır. Mastitis hayvanların sağlığını etkilemekte ve sütteki protein miktarını düşürmektedir. Laktasyondaki hayvanlarda, meme bezlerinin yüzeyleri meme epitel hücreleri ile döşelidir. Meme epitelyum hücrelerinin fonksiyonel durumu kazein ekspresyonu belirlenerek değerlendirilebilir. Total kazein miktarı büyük ölçüde sütün kalitesini yansıtmaktadır. Literatürde süt kalitesi ve mastitiste kazein ekspresyonu hakkında bir bilgi bulunmamaktadır. Bu çalışma, mastitis ile meme epitelyum hücrelerinde α-kazein ekspresyonu arasında bir ilişki bulunup bulunmadığını belirlemek amacıyla yapılmıştır. Lipopolisakkarit (LPS) ile oluşturulmuş mastitis süresince meme epitelyum hücrelerinde in vitro kazein ekspresyonunu değerlendirebilmek amacıyla sığır α-kazeinine karşı antikor üreten hibridoma hücre hattı hazırlanmıştır. Elde edilen sonuçlar, α-kazein ekspresyonunun LPS stimulasyonu altında anlamlı derecede değişmediğini göstermiştir. Bu çalışma sığır meme epitel hücrelerinde α-kazein ekspresyonunu belirlemek amacıyla yararlı olmuştur. Aynı zamanda, sonuçlar mastitis süresince α-kazein ekspresyon seviyesinin stabil olduğunu göstermiştir.

Development of a Monoclonal Antibody Against Bovine α-casein to Evaluate Functional Status of Mammary Epithelial cells During Mastitis

Mastitis is a widespread disease in dairy animals and causes huge economic losses around the globe. Mastitis affects the health of animals and also reduces the protein content in the milk. In lactating animals, the surface of mammary gland is coated and lined with mammary epithelial cells (MECs). The functional status of MECs can be evaluated by determining the expression of caseins. The total content of caseins can largely reflect the quality of milk. In the previous literature, there is no general information about the milk quality and expressions of casein in mastitis. The current study was designed to determine if there is any correlations between mastitis and α-casein expression in MECs. We prepared a hybridoma cell line that produces antibody against bovine α-casein to evaluate the casein expression status in vitro on the MECs during lipopolysaccharide (LPS)-induced mastitis. The results showed that the expression of α-casein has not changed significantly under the stimulation of LPS. Our study established a useful tool to determine the expression profile of α-casein in bovine MECs. Furthermore, the result indicated that the expression level of α-casein remained stable during mastitis.

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  • 1. cakebread J, Hodgkinson A, Wallace O, callaghan M, Hurford D, Wieliczko R, Harris P, Haigh B: Bovine milk derived skimmed milk powder and whey protein concentrate modulates Citrobacter rodentium shedding in the mouse intestinal tract. PeerJ, 6:e5359, 2018. DOI: 10.7717/peerj.5359
  • 2. Belloque J, Ramos M: Determination of the casein content in bovine milk by 31P-NMR. J Dairy Res, 69, 411-418, 2002. DOI: 10.1017/ S0022029902005630
  • 3. castillo DS, cassola A: Novel sensitive monoclonal antibody based competitive enzyme-linked immunosorbent assay for the detection of raw and processed bovine beta-casein. PLoS One, 12(7):e0182447, 2017. DOI: 10.1371/journal.pone.0182447
  • 4. Liao Y, Weber D, Xu W, Durbin-Johnson BP, Phinney BS, Lonnerdal B: Absolute quantification of human milk caseins and the whey/casein ratio during the first year of lactation. J Proteome Res, 16, 4113-4121, 2017. DOI: 10.1021/acs.jproteome.7b00486
  • 5. Newsome GA, Scholl PF: Quantification of allergenic bovine milk αS1casein in baked goods using an intact (1)(5)N-labeled protein internal standard. J Agric Food Chem, 61, 5659-5668, 2013. DOI: 10.1021/jf3015238
  • 6. Ishii H, Nakamura T, Higuchi M, Mamada A, Fukushima M, Urashima T, Arai I: The expression changes of casein mRNAs in mammary epithelial cells recovered from bovine milk during the lactation period. AsianAustralas J Anim Sci, 20, 983-988, 2007. DOI: 10.5713/ajas.2007.983
  • 7. Pujuguet P, Radisky D, Levy D, Lacza c, Bissell MJ: Trichostatin A inhibits β-casein expression in mammary epithelial cells. J Cell Biochem, 83, 660-670, 2001. DOI: 10.1002/Jcb.1260
  • 8. Autran cA, kellman BP, kim JH, Asztalos E, Blood AB, Spence EcH, Patel AL, Hou J, Lewis NE, Bode L: Human milk oligosaccharide composition predicts risk of necrotising enterocolitis in preterm infants. Gut, 67, 1064-1070, 2018. DOI: 10.1136/gutjnl-2016-312819
  • 9. Yamada A, Sakurai T, Ochi D, Mitsuyama E, Yamauchi k, Abe F: Novel angiotensin I-converting enzyme inhibitory peptide derived from bovine casein. Food Chem, 141, 3781-3789, 2013. DOI: 10.1016/j.foodchem. 2013.06.089
  • 10. Hatzoglou A, Bakogeorgou E, Hatzoglou c, Martin PM, castanas E: Antiproliferative and receptor binding properties of α- and β-casomorphins in the T47D human breast cancer cell line. Eur J Pharmacol, 310, 217-223, 1996. DOI: 10.1016/0014-2999(96)00339-1
  • 11. Boutinaud M, Herve L, Lollivier V: Mammary epithelial cells isolated from milk are a valuable, non-invasive source of mammary transcripts. Front Genet, 6:323, 2015. DOI: 10.3389/fgene.2015.00323
  • 12. Li N, Ma W, Shen Q, Zhang M, Du Z, Wu c, Niu B, Liu W, Hua J: Reconstitution of male germline cell specification from mouse embryonic stem cells using defined factors in vitro. Cell Death Differ, 2019. DOI: 10.1038/s41418-019-0280-2
  • 13. Lavon Y, Leitner G, Moallem U, klipper E, Voet H, Jacoby S, Glick G, Meidan R, Wolfenson D: Immediate and carryover effects of Gram-negative and Gram-positive toxin-induced mastitis on follicular function in dairy cows. Theriogenology, 76, 942-953, 2011. DOI: 10.1016/j. theriogenology.2011.05.001
  • 14. Rabot A, Wellnitz O, Meyer HHD, Bruckmaier RM: Use and relevance of a bovine mammary gland explant model to study infection responses in bovine mammary tissue. J Dairy Res, 74, 93-99, 2007. DOI: 10.1017/ S0022029906002147
  • 15. Wu T, Wang c, Ding L, Shen Y, cui H, Wang M, Wang H: Arginine relieves the inflammatory response and enhances the casein expression in bovine mammary epithelial cells induced by lipopolysaccharide. Mediators Inflamm, 2016:9618795, 2016. DOI: 10.1155/2016/9618795
  • 16. Schmitz S, Pfaffl MW, Meyer HHD, Bruckmaier RM: Short-term changes of mRNA expression of various inflammatory factors and milk proteins in mammary tissue during LPS-induced mastitis. Domest Anim Endocrinol, 26, 111-126, 2004. DOI: 10.1016/j.domaniend.2003.09.003
  • 17. Brenaut P, Lefevre L, Rau A, Laloe D, Pisoni G, Moroni P, Bevilacqua c, Martin P: contribution of mammary epithelial cells to the immune response during early stages of a bacterial infection to Staphylococcus aureus. Vet Res, 45:16, 2014. DOI: 10.1186/1297-9716-45-16
  • 18. cubeddu T, cacciotto c, Pisanu S, Tedde V, Alberti A, Pittau M, Dore S, cannas A, Uzzau S, Rocca S, Addis MF: Cathelicidin production and release by mammary epithelial cells during infectious mastitis. Vet Immunol Immunopathol, 189, 66-70, 2017. DOI: 10.1016/j.vetimm.2017.06.002
  • 19. kolb AF, Huber Rc, Lillico SG, carlisle A, Robinson cJ, Neil c, Petrie L, Sorensen DB, Olsson IAS, Whitelaw cBA: Milk lacking α-casein leads to permanent reduction in body size in mice. PLoS One, 6(7):e21775, 2011. DOI: 10.1371/journal.pone.0021775
  • 20. Masoodi TA, Shafi G: Analysis of casein alpha S1 & S2 proteins from different mammalian species. Bioinformation, 4, 430-435, 2010.
  • 21. Sharma N, kim JH, Sodhi SS, Luong DH, kim SW, Oh SJ, Jeong Dk: Differentiation dynamics of mammary epithelial stem cells from Korean holstein dairy cattle under ECM-free conditions. J Biomol Struct Dyn, 33, 2633-2654, 2015. DOI: 10.1080/07391102.2014.1003197
  • 22. German T, Barash I: Characterization of an epithelial cell line from bovine mammary gland. In Vitro Cell Dev Biol Anim, 38, 282-292, 2002. DOI: 10.1290/1071-2690(2002)038<0282:COAECL>2.0.CO;2
  • 23. Huynh HT, Robitaille G, Turner JD: Establishment of bovine mammary epithelial cells (MAC-T): An in vitro model for bovine lactation. Exp Cell Res, 197, 191-199, 1991. DOI: 10.1016/0014-4827(91)90422-Q
  • 24. Smith cJ, Osborn AM: Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology. FEMS Microbiol Ecol, 67, 6-20, 2009. DOI: 10.1111/j.1574-6941.2008.00629.x
  • 25. Boehmer JL, Ward JL, Peters RR, Shefcheck kJ, McFarland MA, Bannerman DD: Proteomic analysis of the temporal expression of bovine milk proteins during coliform mastitis and label-free relative quantification. J Dairy Sci, 93, 593-603, 2010. DOI: 10.3168/jds.2009-2526
  • 26. chiba T, Maeda T, kimura S, Morimoto Y, Sanbe A, Ueda H, kudo k: Inhibitory effect of fluvoxamine on β-casein expression via a serotoninindependent mechanism in human mammary epithelial cells. Eur J Pharmacol, 766, 56-62, 2015. DOI: 10.1016/j.ejphar.2015.09.038
  • 27. Zhao XW, Yang YX, Huang DW, cheng GL, Zhao HL: Comparative proteomic analysis of proteins expression changes in the mammary tissue of cows infected with Escherichia coli mastitis. J Vet Sci, 16, 253-263, 2015. DOI: 10.4142/jvs.2015.16.3.253
Kafkas Üniversitesi Veteriner Fakültesi Dergisi-Cover
  • ISSN: 1300-6045
  • Yayın Aralığı: Yılda 6 Sayı
  • Başlangıç: 1995
  • Yayıncı: Kafkas Üniv. Veteriner Fak.
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