title: Goat uterine DBA+ leukocytes differentiation and cytokines expression respond differently to cloned versus fertilized embryos.
abstract: High rate of fetal mortality in ruminant somatic cell nuclear transfer (SCNT) pregnancies is due, at least in part, to immune-mediated abortion of fetuses. In the present study, goat uterine leukocytes were isolated by Dolichos biflorus agglutinin (DBA) coated magnetic beads, and with majority being were CD56+CD16- in phenotype with low levels of perforin and Granzyme B expression. The responses of the isolated cells to SCNT and in vitro fertilization (IVF) embryos conditioned mediums containing hormone steroids were compared by measuring their phenotype and cytokines expression. The results showed there was a 2-fold increase in the numbers of isolated uterine leukocytes after incubation with different conditioned mediums for 120 h. However, significantly lower percentage and absolute numbers of uterine CD56+CD16- leukocytes incubated with SCNT conditioned mediums were detected as compared with those incubated with IVF conditioned mediums (P < 0.05). The group treated with progesterone (P4) or the combination of P4 and 17β-estradiol (E2) were associated with significantly higher percentage and absolute numbers of CD56+CD16- cells as compared with those treated with E2 alone (P < 0.05). Furthermore, in the presence of steroids, the isolated leukocytes incubated with SCNT conditioned mediums associated with greater levels of IFN-γ secretion and expression, as well as lesser levels of VEGF, as compared with those treated with IVF conditioned mediums (P < 0.05). In conclusion, this study demonstrates that SCNT embryos have a profound effect on the phenotype expression of goat uterine DBA+ leukocytes, as well as the secretion and expression of IFN-γ and VEGF by these cells in vitro.
authors: College of Veterinary Medicine of Northwest A & F University, Yangling, 712100, China.
authors: Lijuan
authors: L
authors: Qin
authors: College of Veterinary Medicine of Northwest A & F University, Yangling, 712100, China.
authors: Mingzhu
authors: M
authors: Lei
authors: College of Veterinary Medicine of Northwest A & F University, Yangling, 712100, China.
authors: Dandan
authors: D
authors: Zhao
authors: College of Veterinary Medicine of Northwest A & F University, Yangling, 712100, China.
authors: Aihua
authors: A
authors: Wang
authors: College of Veterinary Medicine of Northwest A & F University, Yangling, 712100, China.
authors: Yaping
authors: Y
authors: Jin
authors: College of Veterinary Medicine of Northwest A & F University, Yangling, 712100, China.
authors: Xuefeng
authors: X
authors: Qi
mesh_terms: D000373:Agglutinins / Q000378:metabolism*; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D016207:Cytokines / Q000378:metabolism*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000378:metabolism; D004717:Endometrium / Q000166:cytology; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D006041:Goats; D016130:Immunophenotyping; D007962:Leukocytes / Q000166:cytology* / Q000378:metabolism*; D010641:Phenotype; D011987:Receptors, Steroid / Q000378:metabolism; D014599:Uterus
chemical_list: D000373:Agglutinins; D016207:Cytokines; D011987:Receptors, Steroid
references: Domest Anim Endocrinol. 2012 May;42(4):210-9
references: 22226936
references: Theriogenology. 2013 Aug;80(3):228-33
references: 23743066
references: Placenta. 2012 May;33(5):379-86
references: 22357194
references: J Anim Sci. 2007 Mar;85(13 Suppl):E32-5
references: 17040952
references: Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5681-6
references: 19307558
references: J Anim Sci. 2013 Jul;91(7):3002-13
references: 23798511
references: Nat Biotechnol. 1999 May;17(5):456-61
references: 10331804
references: EMBO Mol Med. 2013 Sep;5(9):1415-30
references: 23853117
references: Immunol Rev. 2006 Dec;214:161-85
references: 17100884
references: Methods Mol Biol. 2010;612:465-503
references: 20033660
references: Reprod Fertil Dev. 2012;24(4):588-98
references: 22541547
references: Reprod Biol. 2012 Nov;12(3):309-15
references: 23153702
references: Placenta. 2012 Feb;33 Suppl:S99-S104
references: 22000472
references: Biol Reprod. 2011 Feb;84(2):266-78
references: 20881316
references: Nature. 2005 Aug 4;436(7051):709-13
references: 16079848
references: Placenta. 2008 Mar;29 Suppl A:S108-10
references: 18281092
references: Res Vet Sci. 2002 Apr;72(2):115-21
references: 12027591
references: Genet Mol Res. 2010;9(1):309-23
references: 20198587
references: J Reprod Immunol. 2008 Oct;79(1):18-25
references: 18771804
references: Blood. 2006 Jul 1;108(1):218-27
references: 16527888
references: Biol Reprod. 2002 Jul;67(1):55-63
references: 12079999
references: J Reprod Immunol. 2011 Jun;90(1):105-10
references: 21632120
references: Nat Commun. 2013;4:1479
references: 23403580
references: Biol Reprod. 2012 Oct;87(4):81
references: 22875907
references: Nat Commun. 2014;5:3359
references: 24577131
references: J Immunol. 2004 Dec 1;173(11):6547-63
references: 15557145
references: J Anim Sci. 2007 Mar;85(13 Suppl):E30-1
references: 17322123
references: Biol Reprod. 2012 Nov;87(5):124
references: 23018184
references: Hum Reprod. 2012 Jan;27(1):200-9
references: 22064648
references: Anim Sci J. 2013 Mar;84(3):191-9
references: 23480698
references: Trends Immunol. 2001 Nov;22(11):633-40
references: 11698225
references: PLoS One. 2013;8(6):e65379
references: 23799010
references: Reprod Biol Endocrinol. 2010;8:103
references: 20735829
references: J Immunol. 2010 Apr 1;184(7):3424-32
references: 20194719
references: Theriogenology. 2000 Nov 1;54(8):1207-14
references: 11192179