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Background  The aim of the present study was to evaluate  CCL8  and  CXCL10  expression and its regulatory mechanism in peripheral blood leukocytes (PBLs) at the time of maternal recognition in cows. Blood samples were collected on 14, 15, 16, 17 and 18 d after artificial insemination (AI). Based on the day of return of estrus, cows were divided into three groups, pregnant ( n  = 5), early embryonic mortality (EEM;  n  = 5) and late embryonic mortality (LEM;  n  = 5). The gene expression levels in PBLs were assessed with quantitative real-time reverse transcription PCR.    Results  The expression of  CCL8  and  CXCL10  mRNA in PBLs gradually increased from 14 to 18 d of pregnant cows and significant differences were observed on 18 d ( P  < 0.05), whereas no significant changes were observed both in EEM and LEM cows. Interferon-stimulated protein 15 kDa ( ISG15 ), myxovirus-resistance gene ( MX ) 1 and  MX2  mRNA expression in PBLs increased from 14 to 18 d which was significant on 18 d of pregnant cows as well as in LEM cows ( P  < 0.05), but no changes were observed in EEM cows. To determine whether the expression of  CCL8  and  CXCL10  in PBLs was regulated by pregnancy-related substances or not, expression level was assessed after exposure to interferon-τ (IFNT) and CCL16. Monocytes, granulocytes and lymphocytes were obtained using density-gradient centrifugation and flow cytometry. The addition of IFNT (100 ng/mL) and CCL16 (100 ng/mL) to cultured PBLs increased the expression of  CCL8  and  CXCL10  mRNA ( P  < 0.05). The expression of  ISG15 ,  MX1  and  MX2  mRNA in PBLs was also stimulated by IFNT and CCL16 ( P  < 0.05).    Conclusions  The expression of  CCL8  and  CXCL10  genes increased in PBLs during early pregnancy. Since IFNT stimulated  CCL8  and  CXCL10  expression in cultured PBLs, the increase of CCL8 and CXCL10 might be pregnancy-dependent events. The expression of both  CCL8  and  CXCL10  in PBLs was stimulated by CCL16 as well as IFNT, suggesting a chemokine interaction that at least includes CCL8, CXCL10 and CCL16, and may play a role in regulating maternal recognition in cows.

Gene expression of CCL8 and CXCL10 in peripheral blood leukocytes during early pregnancy in cows