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Am J Transl Res 2013;5(1):103-115
Original Article
Differential effect of exogenous interleukin-10 versus glucocorticoids
on gene expression and pro-inflammatory cytokine release by
polymorphonuclear leukocytes and monocytes of the newly born
Dennis Davidson, Hardik Patel, Ana C Degoy, Irina Gershkovich, Ivana Vancurova, Veronika Miskolci
Division of Neonatal-Perinatal Medicine, Stony Brook Long Island Children’s Hospital, Stony Brook, NY 11794,
USA; The Feinstein Institute for Medical Research, Manhasset, NY 11030, USA; Department of Biology, St John’s
University, New York, NY 11439, USA
Received October 6, 2012; Accepted November 18, 2012; Epub January 21, 2013; Published January 30, 2013
Abstract: Bronchopulmonary dysplasia (BPD) is one of the most common causes of mortality and morbidity in
neonatal intensive care units. Persistent inflammation, with an abnormal influx of polymorphonuclear leukocytes
(PMNs) followed by monocytes (MONOs), occurs early in the pathogenesis of BPD. Anti-inflammatory therapy with
better efficacy and safety than dexamethasone (DEX) is needed. In the present study we determined cell-specific
gene expression and cytokine release in response to glucocorticoids versus interleukin-10 (IL-10). Subsequently,
we hypothesized that the insensitivity of MONOs to DEX was associated with a failure of the glucocorticoid
receptor to translocate to the nucleus. PMNs and MONOs were isolated from umbilical cord blood at birth, and
pretreated with PBS vehicle, IL-10 or glucocorticoids prior to endotoxin (LPS)-stimulation for 4 and 18h. Genome-
wide gene expressions were determined by microarray and validated by RT-qPCR. Interleukin 8 release in cell
culture supernatant was measured by ELISA. To examine the mechanism of monocyte insensitivity to
glucocorticoids, nuclear translocation of the glucocorticoid receptor was determined by Western blots. MONOs
had 6 times the number of genes changing expression with IL-10 compared to PMNs at 4h. DEX at the
therapeutic level for neonates with BPD had no effect on gene expression in MONOs. The order of potency for
inhibition of interleukin-8 release from MONOs was IL-10 >betamethasone >dexamethasone and hydrocortisone.
Glucocorticoid potency in MONOs was directly related to glucocorticoid receptor translocation to nucleus. Gene
expression profiling for IL-10 versus glucocorticoids indicates there may be major differences in therapeutic
efficacy for BPD. (AJTR1210001).
Keywords: Bronchopulmonary dysplasia, monocytes, polymorphonuclear leukocytes, gene expression
microarray, dexamethasone
Address correspondence to: Dr. Dennis Davidson, Division of Neonatal-Perinatal Medicine, HSC T-11, 060,
Stony Brook Long Island Children’s Hospital, Stony Brook, NY 11794, USA. Phone: 631-444-7653; Fax: 631-444-
8968; E-mail: dennis.davidson@stonybrookmedicine.edu
Original Article
Differential effect of exogenous interleukin-10 versus glucocorticoids
on gene expression and pro-inflammatory cytokine release by
polymorphonuclear leukocytes and monocytes of the newly born
Dennis Davidson, Hardik Patel, Ana C Degoy, Irina Gershkovich, Ivana Vancurova, Veronika Miskolci
Division of Neonatal-Perinatal Medicine, Stony Brook Long Island Children’s Hospital, Stony Brook, NY 11794,
USA; The Feinstein Institute for Medical Research, Manhasset, NY 11030, USA; Department of Biology, St John’s
University, New York, NY 11439, USA
Received October 6, 2012; Accepted November 18, 2012; Epub January 21, 2013; Published January 30, 2013
Abstract: Bronchopulmonary dysplasia (BPD) is one of the most common causes of mortality and morbidity in
neonatal intensive care units. Persistent inflammation, with an abnormal influx of polymorphonuclear leukocytes
(PMNs) followed by monocytes (MONOs), occurs early in the pathogenesis of BPD. Anti-inflammatory therapy with
better efficacy and safety than dexamethasone (DEX) is needed. In the present study we determined cell-specific
gene expression and cytokine release in response to glucocorticoids versus interleukin-10 (IL-10). Subsequently,
we hypothesized that the insensitivity of MONOs to DEX was associated with a failure of the glucocorticoid
receptor to translocate to the nucleus. PMNs and MONOs were isolated from umbilical cord blood at birth, and
pretreated with PBS vehicle, IL-10 or glucocorticoids prior to endotoxin (LPS)-stimulation for 4 and 18h. Genome-
wide gene expressions were determined by microarray and validated by RT-qPCR. Interleukin 8 release in cell
culture supernatant was measured by ELISA. To examine the mechanism of monocyte insensitivity to
glucocorticoids, nuclear translocation of the glucocorticoid receptor was determined by Western blots. MONOs
had 6 times the number of genes changing expression with IL-10 compared to PMNs at 4h. DEX at the
therapeutic level for neonates with BPD had no effect on gene expression in MONOs. The order of potency for
inhibition of interleukin-8 release from MONOs was IL-10 >betamethasone >dexamethasone and hydrocortisone.
Glucocorticoid potency in MONOs was directly related to glucocorticoid receptor translocation to nucleus. Gene
expression profiling for IL-10 versus glucocorticoids indicates there may be major differences in therapeutic
efficacy for BPD. (AJTR1210001).
Keywords: Bronchopulmonary dysplasia, monocytes, polymorphonuclear leukocytes, gene expression
microarray, dexamethasone
Address correspondence to: Dr. Dennis Davidson, Division of Neonatal-Perinatal Medicine, HSC T-11, 060,
Stony Brook Long Island Children’s Hospital, Stony Brook, NY 11794, USA. Phone: 631-444-7653; Fax: 631-444-
8968; E-mail: dennis.davidson@stonybrookmedicine.edu
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