Cord Blood Leukocytes Research Articles

Innate Immune Deficiency of Extremely Premature Neonates Can Be Reversed by Interferon-γ

BackgroundBacterial sepsis is a major threat in neonates born prematurely, and is associated with elevated morbidity and mortality. Little is known on the innate immune response to bacteria among extremely premature infants.Methodology/Principal FindingsWe compared innate immune functions to bacteria commonly causing sepsis in 21 infants of less than 28 wks of gestational age, 24 infants born between 28 and 32 wks of gestational age, 25 term newborns and 20 healthy adults. Levels of surface expression of innate immune receptors (CD14, TLR2, TLR4, and MD-2) for Gram-positive and Gram-negative bacteria were measured in cord blood leukocytes at the time of birth. The cytokine response to bacteria of those leukocytes as well as plasma-dependent opsonophagocytosis of bacteria by target leukocytes was also measured in the presence or absence of interferon-γ. Leukocytes from extremely premature infants expressed very low levels of receptors important for bacterial recognition. Leukocyte inflammatory responses to bacteria and opsonophagocytic activity of plasma from premature infants were also severely impaired compared to term newborns or adults. These innate immune defects could be corrected when blood from premature infants was incubated ex vivo 12 hrs with interferon-γ.Conclusion/SignificancePremature infants display markedly impaired innate immune functions, which likely account for their propensity to develop bacterial sepsis during the neonatal period. The fetal innate immune response progressively matures in the last three months in utero. Ex vivo treatment of leukocytes from premature neonates with interferon-γ reversed their innate immune responses deficiency to bacteria. These data represent a promising proof-of-concept to treat premature newborns at the time of delivery with pharmacological agents aimed at maturing innate immune responses in order to prevent neonatal sepsis.

Insulin-Like Growth Factor 2/H19 Methylation at Birth and Risk of Overweight and Obesity in Children

To determine whether aberrant DNA methylation at differentially methylated regions (DMRs) regulating insulin-like growth factor 2 (IGF2) expression in umbilical cord blood is associated with overweight or obesity in a multiethnic cohort. Umbilical cord blood leukocytes of 204 infants born between 2005 and 2009 in Durham, North Carolina, were analyzed for DNA methylation at two IGF2 DMRs by using pyrosequencing. Anthropometric and feeding data were collected at age 1 year. Methylation differences were compared between children >85th percentile of the Centers for Disease Control and Prevention growth charts weight-for-age (WFA) and children ≤ 85th percentile of WFA at 1 year by using generalized linear models, adjusting for post-natal caloric intake, maternal cigarette smoking, and race/ethnicity. The methylation percentages at the H19 imprint center DMR was higher in infants with WFA >85th percentile (62.7%; 95% CI, 59.9%-65.5%) than in infants with WFA ≤ 85th percentile (59.3%; 95% CI, 58.2%-60.3%; P = .02). At the intragenic IGF2 DMR, methylation levels were comparable between infants with WFA ≤ 85th percentile and infants with WFA >85th percentile. Our findings suggest that IGF2 plasticity may be mechanistically important in early childhood overweight or obese status. If confirmed in larger studies, these findings suggest aberrant DNA methylation at sequences regulating imprinted genes may be useful identifiers of children at risk for the development of early obesity.

Short Fetal Leukocyte Telomere Length and Preterm Prelabor Rupture of the Membranes

BackgroundRupture of the fetal membranes is a common harbinger of imminent labor and delivery. Telomere shortening is a surrogate for oxidative stress (OS) and senescence. Fetal leukocyte and placental membrane DNA telomere lengths were evaluated to determine their association with preterm prelabor rupture of the membranes (pPROM) or spontaneous preterm births with intact membranes (PTB), compared to term birth.MethodsTelomere lengths were quantified in cord blood leukocytes (n = 133) from three major groups: 1) pPROM (n = 28), 2) PTB (n = 69) and 3) uncomplicated full term births (controls, n = 35), using real-time quantitative PCR. Placental membrane specimens (n = 18) were used to correlate fetal leukocyte and placental telomere lengths. Telomere length differences among the groups were analyzed by ANOVA. Pearson correlation coefficients determined relationships between leukocyte and placental membrane telomere lengths.ResultsIn pregnancies with intact membranes, fetal leukocyte telomere length was inversely proportional to gestational age. The mean telomere length decreased as gestation progressed, with the shortest at term. pPROM had telomere lengths (9962±3124 bp) that were significantly shorter than gestational age-matched PTB (11546±4348 bp, p = 0.04), but comparable to term births (9011±2497 bp, p = 0.31). Secondary analyses revealed no effects of race (African American vs. Caucasian) or intraamniotic infection on telomere length. A strong Pearson's correlation was noted between fetal leukocyte and placental membrane telomere lengths (ρ = 0.77; p<0.01).ConclusionsFetal leukocyte telomere length is reduced in pPROM compared to PTB but is similar to term births. pPROM represents a placental membrane disease likely mediated by OS-induced senescence.

The effects of depression and use of antidepressive medicines during pregnancy on the methylation status of the IGF2 imprinted control regions in the offspring

In utero exposures to environmental factors may result in persistent epigenetic modifications affecting normal development and susceptibility to chronic diseases in later life. We explored the relationship between exposure of the growing fetus to maternal depression or antidepressants and DNA methylation at two differentially methylated regions (DMRs) of the imprinted Insulin-like Growth Factor 2 (IGF2) gene. Aberrant DNA methylation at the IGF2 and neighboring H19 DMRs has been associated with deregulated IGF2 expression, childhood cancers and several chronic diseases during adulthood. Our study population is comprised of pregnant mothers and their newborns (n = 436), as part of the Newborn Epigenetics Study (NEST). A standardized questionnaire was completed and medical record data were abstracted to ascertain maternal depression and antidepressive drug use. DMR methylation levels in umbilical cord blood leukocytes were quantified using pyrosequencing. From the 436 newborns, laboratory data were obtained for 356 individuals at the IGF2 DMRs, and for 411 individuals at the H19 DMRs; about half of each group was African American or Caucasian. While overall no association between depression and methylation profiles was found, we observed a significant hypermethylation of the H19 DMRs in newborns of African American (n = 177) but not Caucasian (n = 168) mothers who reported the use of antidepressive drugs during pregnancy (β = +6.89, p = 0.01). Of note, our data reveal a race-independent association between smoking during pregnancy and methylation at the IGF2 DMR (+3.05%, p = 0.01). In conclusion, our findings suggest a race-dependent response related to maternal use of antidepressants at one of the IGF2 DMRs in the offspring.

Methylation variation at IGF2 differentially methylated regions and maternal folic acid use before and during pregnancy

Folic acid (FA) supplementation before and during pregnancy has been associated with decreased risk of neural tube defects although recent reports suggest it may also increase the risk of other chronic diseases. We evaluated exposure to maternal FA supplementation before and during pregnancy in relation to aberrant DNA methylation at two differentially methylated regions (DMRs) regulating Insulin-like Growth Factor 2 (IGF2) expression in infants. Aberrant methylation at these regions has been associated with IGF2 deregulation and increased susceptibility to several chronic diseases. Using a self-administered questionnaire, we assessed FA intake before and during pregnancy in 438 pregnant women. Pyrosequencing was used to measure methylation at two IGF2 DMRs in umbilical cord blood leukocytes. Mixed models were used to determine relationships between maternal FA supplementation before or during pregnancy and DNA methylation levels at birth. Average methylation at the H19 DMR was 61.2%. Compared to infants born to women reporting no FA intake before or during pregnancy, methylation levels at the H19 DMR decreased with increasing FA intake (2.8%, p=0.03, and 4.9%, p=0.04, for intake before and during pregnancy, respectively). This methylation decrease was most pronounced in male infants (p=0.01). Methylation alterations at the H19 DMR are likely an important mechanism by which FA risks and/or benefits are conferred in utero. Because stable methylation marks at DMRs regulating imprinted genes are acquired before gastrulation, they may serve as archives of early exposures with the potential to improve our understanding of developmental origins of adult disease.

Abstract 2754: Maternal use of antidepressants in pregnancy is associated with hypermethylation at the IGF2 imprinted control regions in the offspring in a race-dependent fashion

Abstract In utero exposures to environmental factors may result in persistent epigenetic modifications affecting normal development and susceptibility to chronic diseases in later life. We determined whether exposure of the growing fetus to maternal depression or antidepressants is associated with aberrant DNA methylation at two differentially methylated regions (DMRs) of the imprinted Insulin-like Growth Factor 2 (IGF2) gene, namely the DMRs upstream of the imprinted IGF2 promoters (IGF2 DMR) and the DMRs upstream of H19 (H19 DMR). Aberrant DNA methylation at these DMRs has been associated with deregulated IGF2 expression, childhood cancers and several chronic diseases during adulthood. Our study population comprises pregnant mothers and their newborns (n=436), recruited between 2005 and 2008 in Durham, NC. A standardized questionnaire was completed and medical record data were abstracted to ascertain prenatal and periconceptional depression and antidepressive drug use. DMR methylation percentages in umbilical cord blood leukocytes were quantified using pyrosequencing. While an association between depression and methylation profiles was not found, we discovered significant hypermethylation of the H19 DMRs in newborns of African American -but not Caucasian- mothers who were taking antidepressive drugs during pregnancy (β=+6.89, p=0.01). Our findings suggest that changes in the IGF2/H19-domain methylation in response to maternal exposure to antidepressants are race-dependent. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2754. doi:10.1158/1538-7445.AM2011-2754

Glyceraldehyde-3-phosphate dehydrogenase is a reliable internal control in Western blot analysis of leukocyte subpopulations from children

To study differences in the development of immunity, leukocytes from cord blood are often compared with those from adult peripheral blood. Western blot analysis is a common method for detecting proteins. In this study, we investigated the reliability of using different housekeeping proteins (β-actin, β-tubulin, and glyceraldehyde-3-phosphate dehydrogenase [GAPDH]) as internal controls for different leukocyte subpopulations from infants, children, and adults. Our results showed that the expression levels of β-actin and β-tubulin were much lower in cord blood leukocytes than in adult leukocytes, and this expression pattern persisted in children up to 3 years old. Further study revealed that the β-actin expression level in newborns was especially lower in CD14-positive monocytes. However, cord blood and adult peripheral blood monocytes had similar expression levels of β-actin messenger RNA (mRNA). Further experiments showed that posttranslational regulation was responsible for the low β-actin expression level in neonatal monocytes. Thus, researchers should carefully assess the appropriate use of housekeeping gene-encoded proteins as internal standards to normalize samples for comparisons of different leukocyte populations from subjects of different ages. In this study, we determined that GAPDH was a more reliable internal control than others in Western blot analysis for comparing the development of immunity among infants, children, and adults.

Ocena ilościowa limfocytów B i ich subpopulacji we krwi pępowinowej noworodków w zależności od sposobu ukończenia ciąży u ich matek

Aim The aim of the study was quantitative evaluation and determination to what degree the time of delivery causes a change in percentage and number of CD19+ lymphocytes and their subpopulations CD5+, CD23+. Material and Method The study included 72 healthy neonates born in the Perinatology and Gynecology Ward of Silesian Medical University in Zabrze. Considering the duration of pregnancy and the method of delivery, the children were divided in the following groups. Group I: 40 neonates born in term which included 17 neonates born naturally and 23 neonates born by cesarean section, of which 15 with elective indications and 8 with emergency indications. Group II: 32 neonates born prematurely, which included 6 neonates born naturally and 26 neonates born by cesarean section, of which 18 with elective indications and 8 with emergency indications. In the current study the cord blood leukocyte were examined with flow cytometry technique. Results The immunological factors in neonates were analyzed with reference to the time of delivery. It was found that the mean percentage of CD5+ lymphocytes is significantly lower than in neonates born prematurely. Similar differences were shown for all the neonates born by cesarean section and by elective cesarean section. However, the mean percentage of CD23+ lymphocytes was statistically significantly higher in the neonates born in term by cesarean section due to emergency indications than in the neonates born prematurely in the same way. The study showed that the mean number of CD23+ lymphocytes in the neonates born in time by cesarean section with elective indications was significantly statistically lower in the neonates born by cesarean section with emergency indications. Conclusions 1. Time of delivery in cesarean section can result in significant changes in number of CD5+, CD23+ B lymphocytes in neonates. 2. Cesarean section performed with emergency indications can favor an increase of B lymphocytes mean number CD23+ B lymphocytes in neonates born in term.

Prenatal animal contact and gene expression of innate immunity receptors at birth are associated with atopic dermatitis

Cross-sectional studies have suggested that prenatal farm exposures might protect against allergic disease and increase the expression of receptors of the innate immune system. However, epidemiologic evidence supporting the association with atopic dermatitis remains inconsistent. To study the association between prenatal farm-related exposures and atopic dermatitis in a prospective study. We further analyzed the association between the expression of innate immune genes at birth and atopic dermatitis. A total of 1063 children who participated in a birth cohort study, Protection against Allergy-Study in Rural Environments, were included in this study. Doctor diagnosis of atopic dermatitis was reported by the parents from 1 to 2 years of age by questionnaire. Gene expression of Toll-like receptors (TLRs) and CD14 was assessed in cord blood leukocytes by quantitative PCR. Maternal contact with farm animals and cats during pregnancy had a significantly protective effect on atopic dermatitis in the first 2 years of life. The risk of atopic dermatitis was reduced by more than half among children with mothers having contact with 3 or more farm animal species during pregnancy compared with children with mothers without contact (adjusted odds ratio, 0.43; 95% CI, 0.19-0.97). Elevated expression of TLR5 and TLR9 in cord blood was associated with decreased doctor diagnosis of atopic dermatitis. A significant interaction between polymorphism in TLR2 and prenatal cat exposure was observed in atopic dermatitis. Maternal contact with farm animals and cats during pregnancy has a protective effect on the development of atopic dermatitis in early life, which is associated with a lower expression of innate immune receptors at birth.

Experimental Evaluation of Rat Cord Blood as a Source of Stem Cells Stimulating Regeneration of the Bone Tissue

We studied subpopulation structure of rat cord blood and its effects on regeneration of the bone tissue. In cord blood leukocytes, cell fractions with phenotypes CD45+/CD90-, CD45-/CD90+, CD34+/CD45+ were determined. The cells adhered to plastic during culturing and had a fibroblast-like morphology. Transplantation of rat cord blood cells into modeled femoral bone defects stimulated regeneration of the bone tissue and led to recovery of its anatomical integrity, which was confirmed by X-ray examination and histological analysis. No complete recovery of the bone structure was observed in controls (without cell implantation).

The Central Role of Leukotriene B4 in Epstein Barr Virus Activated Innate Immunity In Vitro

Leukotriene B4 (LTB4) is a potent fast acting lipid mediator produced mainly by activated granulocytes and monocytes. Epstein Barr Virus (EBV) is ubiquitous in humans. Primary infection is usually asymptomatic in young children, but in adolescents, who are most often the subjects of the infection, it is followed often by the characteristic syndrome of infectious mononucleosis (IM). We have assessed the contribution of LTB4 to the primary response in EBV infected cord blood leukocyte cultures and found that addition of LTB4 led to inhibition of the EBV induced B lymphocyte proliferation. Some of the symptoms of IM can be regarded as a consequence of the activation of the innate immunity system. Our results indicate that LTB4 may play a pathophysiological role in this process. The overall effect of LTB4 in mononucleosis is hard to predict. The endogenous production of LTB4 in affected IM tissue might be of importance for the host defence but at the same time strengthen the symptoms. Therefore further studies are warranted to elucidate whether LTB4 is involved in the symptomatology of mononucleosis. If so, administration of pharmacological inhibitors of the synthesis or the action of leukotrienes may be beneficial in severe cases. Keywords: leukotriene B4, Epstein-Barr-virus, EBV, innate immunity, infectious mononucleosis, IM, BLT1

FETAL MALNUTRITION

Fetal malnutrition has been associated with maturational defect in cell metabolism. A series of studies on anthropometric and clinical aspects of patients with intrauterine malnutrition was conducted to establish the relationship between intrauterine growth failure and cell function. The 1st study assessed intrauterine growth in 128 cases. 36 infants were found to be small for gestational age and exhibited usual signs of fetal growth retardation. The mothers of these infants were similar with respect to age, weight, height, nutritional patterns, and prior pregnancy histories. The 2nd study determined whether there was a unique pattern for certain cell energy function in leukocytes of mothers and their small-for-gestational age infants at the time of birth, and whether this pattern was the same as that found in infants with severe postnatal protein-calorie malnutrition. The findings showed that the altered pattern of energy metabolism in leukocytes of infants with intrauterine growth retardation was the same as that found in young infants with postnatal protein-calorie malnutrition. The similarity of enzyme pattern in the adenine nucleotide contents of maternal and cord blood leukocytes at time of delivery also supported the interrelationship between energy metabolisms of mothers and their fetuses. The 3rd study suggested that fetal malnutrition may be reflected in placental tissues. These studies may prove useful in developing diagnostic tools for monitoring fetal growth and for establishing criteria for intervention.

Gene expression measurements in the context of epidemiological studies

Gene expression measurements became an attractive tool to assess biological responses in epidemiological studies. However, collection of blood samples poses various technical problems. We used gene expression data from two epidemiological studies to evaluate differences between sampling methods, comparability of two methods for measuring RNA levels and stability of RNA samples over time. For the PARSIFAL study, PBLC of 1155 children were collected using EDTA tubes in two countries. In the PASTURE study, tubes containing RNA-stabilizing solutions (PAXgene) Blood RNA Tubes; PreAnalytiX) were used to collect cord blood leucocytes of 982 children in five countries. Real-time PCR (conventional single tube assay and high-throughput low density arrays) was used to quantify expression of various innate immunity genes. In 77 PARSIFAL samples, gene expression was measured repeatedly during prolonged storage. In PARSIFAL (EDTA tubes) the median RNA yield after extraction significantly differed between the two centres (70 and 34 ng/microl). Collecting blood into an RNA-stabilizing solution markedly reduced differences in RNA yield in PASTURE (range of medians 91-107 ng/microl). The agreement [Spearman rank correlation (r)] between repeated measurements of gene expression decreased with increasing storage time [e.g., for CD14: r (first/second measurement) = 0.35; r (first/third measurement) = 0.03]. RNA levels measured with either the conventional method or low-density arrays were comparable (r > 0.9). Collecting blood samples into tubes containing an RNA-stabilizing solution increases RNA yield and reduces its variability. Long-term storage of samples may lead to RNA degradation, requiring special attention in longitudinal studies.

Abstract B45: Prenatal exposure to polycyclic aromatic hydrocarbons (PAH), PAH-DNA adducts and genomic DNA methylation in cord blood

Abstract B45 Polycyclic aromatic hydrocarbons (PAHs) are widespread organic carcinogenic pollutants. When metabolized in vivo, PAHs form reactive diol epoxides that bind to DNA at guanine residues forming adducts. PAH exposure has also been associated with alterations in genomic cytosine methylation. Both PAH-DNA adducts and altered DNA methylation have been associated with increased cancer risk. We sought to explore the relationship between prenatal PAH exposure, global DNA methylation and PAH-DNA adducts in a New York City birth cohort. Cord blood was collected from deliveries of non-smoking, African Americans and Dominicans, ages 18-35, residing in N. Manhattan and S. Bronx. Personal air monitors measured PAH during their 8th month of pregnancy. We measured benzo[a]pyrene (B[a]P)-DNA adducts (which serve as a proxy for PAH-DNA adducts) in cord blood using high-performance liquid chromatography-fluorescence. We measured genomic DNA methylation in cord blood leukocytes using the MethylampTM global DNA methylation quantification kit. Demographic and epidemiologic risk factors were collected prospectively, including maternal age, ethnicity, parity, and prenatal tobacco smoke exposure, and child’s gender We found that prenatal PAH exposure was associated global DNA hypomethylation such that average methylation among those in the highest quartile of prenatal PAH exposure was 1.17 ng/100 ng total DNA as compared 1.83 ng/100 ng total DNA in the lowest quartile (p &amp;lt; 0.01). Conversely, those with detectable PAH-DNA adducts had higher levels of DNA methylation (1.43 ng/100 ng total DNA) as compared to those with non-detectable adducts (1.16 ng/100 ng total DNA) (p = 0.02). Using multiple variable linear regression, PAH exposure was independently associated with hypomethylation (average difference in methylation among those in the highest vs. lowest quartile of exposure = -0.42, 95% CI: -0.75, -0.09) and the presence of DNA adducts was associated with hypermethyaltion (average difference in methylation among those with detectable vs. non-detectable adducts = 0.28, 95% CI: 0.05, 0.49). Adjusting for potential confounders did not significantly alter these independent associations, indicating that the lowest level of global DNA methylation was found among those with non-detectable adducts and high PAH exposure, and the highest level of DNA methylation was found among those with detectable adducts and low PAH exposure. While the mechanism by which prenatal PAH exposure results in DNA adducts is well understood, the way exposure may impact methylation is not known. These data suggest that in the presence of DNA adducts, PAH exposure is associated with a reduction in methylation, but when adducts are non-detectable, the same PAH exposure is associated with an even larger methylation reduction. It is possible that the presence of adducts may be partially preventing PAH compounds (or metabolites) from accessing the DNA, leading to less of a change in methylation. It is also possible that the shift in global methylation from PAH exposure may prevent adduct formation. These speculations as well as the associations observed in this analysis require confirmation in other settings/populations. Because methylation changes are potentially reversible, it is possible that if these associations are confirmed, methylation may serve as a target for intervention. Citation Information: Cancer Prev Res 2008;1(7 Suppl):B45.

Fetal-Maternal HLA-A and – B Discordance is Associated with Placental RNase Expression and Anti-HIV-1 Activity

The incidence of maternal-to-fetal human immunodeficiency virus type 1 (HIV-1) transmission is 25-30% in absence of antiretroviral therapy, and is inversely associated with Human leukocyte antigens (HLA) class-I discordance. Based on our earlier report that mixed lymphocyte reactions (MLR) induce a ribonuclease (RNase) that inhibits HIV-1 replication, we proposed that maternal-fetal alloantigen stimulation activates factors that protect the fetus against vertically-transmitted infections. We investigate here whether the degree of mother-infant HLA discordance associates with the ability to produce anti-HIV-1 alloantigen-stimulated factor (ASF), and affects placental RNases. We also determine whether such HLA association is influenced by the mother's HIV-1 status. Paired maternal and cord blood leukocytes were tested for the induction of ASF by MLR, and typed for HLA-A and -B. The placentas were tested for mRNA expression of three RNases. Neonate anti-mother, but not mother anti-neonate MLR generated supernatants with anti-HIV-1 activity, that was associated with HLA class I discordance. This HLA association was not seen in the HIV-infected cohort. HLA class I discordance was also associated with expression of placental RNase 1. Our findings are consistent with the hypothesis that HLA class I discordance induces expression of RNases in the placenta that contribute to innate host resistance to HIV-1 and other viral infections.

Using allogeneic stem cell/T-cell grafts to cure hematologic malignancies

Background: Allogeneic stem cell and T-cell-based therapies are widely used in the treatment of hematologic malignancies and can treat or cure otherwise refractory disease. However, in spite of major advances in the understanding and practice of allogeneic hematopoietic cell transplantation (HCT), several important challenges remain. Objective: Here the authors review the use of allogeneic HCT and T-cell-based therapy, with the goal of providing an overview of the uses and limitations of this approach as well as a survey of areas of active research. Methods: The authors review and summarize recent publications and expert opinions in the field of allogeneic HCT, along with a brief historical perspective, with a focus on challenges and recent advances in the field. Results/conclusion: Present areas of research include efforts to expand the donor pool through the use of umbilical cord blood and human leukocyte antigen-haploidentical donors, the use of reduced-intensity conditioning regimens, which allow treatment of previously ineligible patients, enhancement of immune reconstitution after transplantation, more effective prevention and treatment of acute and chronic graft-versus-host disease, and the augmentation of the immunologic graft-versus-tumor response and its uncoupling from deleterious graft-versus-host alloreactivity.

The developmental changes in mitochondrial DNA content per cell in human cord blood leukocytes during gestation

The mitochondrial DNA (mtDNA) amount in cells as the basis for mitochondrial energy generating system, which produces ATP, plays an important role in the fetal development and postnatal morbidity. Isolated human cord blood leukocytes (HCBL) contribute very little to the overall metabolic turnover, but they may serve as easily available marker cells for the study of the mtDNA amount changes in cord blood during fetal development. The aim of our study was to analyze the mtDNA amount in HCBL. HCBL were isolated from cord blood samples of 107 neonates born between the 25th and 41st week of gestation. The mtDNA amount was analyzed by the real-time PCR method. The significant negative correlations were found between the relative mtDNA amount in HCBL and gestational age (r = -0.54, p<0.01) and birth weight (r = -0.43, p<0.01), respectively. The results revealed that the mtDNA content per cell decreases in HCBL with progressing fetal development. This may be explained by gradual shift of the hematopoiesis from fetal liver to bone marrow during the second half of pregnancy presumably accompanied by decreasing cell volume of HCBL as it was shown similarly in red blood cells.

Intrauterine therapy with cytomegalovirus hyperimmunoglobulin for a fetus congenitally infected with cytomegalovirus

Reported herein is a case of hydrops fetalis in which the cord blood expressed cytomegalovirus (CMV) antigen. Fetal ascites was removed from an infected fetus with hydrops in utero and 2.5 g CMV hyperimmunoglobulin was administered into the fetal abdominal cavity at 28 weeks gestation. After immunoglobulin administration, fetal ascites vanished, preload index of the inferior vena cava decreased and platelet count of the infant increased. However, despite intrauterine therapy and intensive neonatal care, the infant died soon after birth. The expression of CMV antigen in the nucleus of polymorphonuclear leukocytes in fetal cord blood indicated that the fetal hydrops was caused by CMV infection. When symptomatic CMV infection of a fetus is suspected from serological and ultrasound findings, further examinations should be performed for the diagnosis. Intrauterine immunoglobulin therapy could be one of the therapeutic options for the affected fetus.

Cytokine profiles of cord and adult blood leukocytes: differences in expression are due to differences in expression and activation of transcription factors

BackgroundStem cell transplantation as therapy for hematological disorders is often hampered by severe graft-versus-host-disease. This may be reduced by umbilical cord blood transplantation, an effect that has been attributed to qualitative differences between neonatal and adult T cells. We compared levels of secreted proteins and cytokine mRNA induced in cord blood leukocytes (CBL) and adult blood leukocytes (ABL) by various stimuli.ResultsWhile interleukin-2 (IL-2) levels were similar in CBL and ABL, there was less induction of the Th1 cytokine interferon-γ in CBL. Production of the Th2 cytokines IL-4, IL-5, and IL-13 and the hematopoietic cytokine IL-3 was much lower in CBL versus ABL after T-cell receptor-mediated stimulation, whereas production of GM-CSF was comparable in the 2 cell types. The lower levels of Th1 and Th2 cytokines were maintained in CBL during a 4-day time-course study, while after 12 hours IL-3 and GM-CSF reached in CBL levels similar to those in ABL. For all cytokines except IFNγ, the IC50 values for inhibition by cyclosporin A were similar in ABL and CBL. In contrast, there was less expression and activation of transcription factors in CBL. Activation of NF-κB by TPA/ionomycin was detected in ABL but not CBL. Furthermore, there was less expression of the Th subset-specific transcription factors T-bet and c-maf in CBL versus ABL, whereas GATA-3 expression was similar. Expression of T-bet and c-maf correlated with expression of the Th1 and Th2 cytokines, respectively. Time course experiments revealed that T-bet expression was stimulated in both cell types, whereas c-maf and GATA-3 were induced only in ABL.ConclusionThe diminished capability of CBL to synthesize cytokines is probably due to decreased activation of NF-κB, whereas differences in Th subsets are due to differences in regulation of Th lineage-specific transcriptions factors. We propose that the reduced incidence and severity of GvHD after allogeneic transplantation of umbilical CB cells is due to lesser activation of specific transcription factors and a subsequent reduction in production of certain cytokines.

Comparison of maternal and cord blood nucleated red blood cell count between pre‐eclamptic and healthy women

The aim of this study was to evaluate the influence of pre-eclampsia on the cord and maternal nucleated red blood cell (NRBC) count. Immediately after delivery, 1 mL of maternal venous blood and 1 mL of cord blood from 50 pre-eclamptic and 150 healthy pregnant women were collected separately in tubes containing 1.5 mg ethylene diamine tetra-acetic acid. Blood smears were prepared and stained using the Giemsa method. The number of NRBC per 100 leukocytes in maternal and cord blood was counted and compared between the two groups using SPSS software package for Windows. Any correlation of the NRBC count in maternal and umbilical cord blood was also evaluated. P-values < 0.05 were considered significant. The mean (+/-SD) NRBC per 100 white blood cell (WBC) level in cord blood of newborns in the pre-eclamptic group (18.2 +/- 31.8, range 0-142) was significantly greater than in the control group (6.2 +/- 8.1, range 0-36). Low birth weight and intrauterine growth restriction showed a statistically significant relationship with abnormal NRBC count in pre-eclamptic patients. A significant correlation was found between the maternal and cord blood NRBC count in the pre-eclamptic group. Fetal response to utero-placental insufficiency in pre-eclampsia leads to elevated NRBC in the cord blood, particularly in the presence of low birth weight and intrauterine growth restriction. The positive correlation between maternal and cord blood NRBC counts in pre-eclamptic patients indicates that maybe the hypoperfused placenta plays a role in the correlated alteration of the maternal and fetal NRBC count.