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The impact of maternal HIV infection on cord blood lymphocyte subsets and cytokine profile in exposed non-infected newborns.

Borges-Almeida E, Milanez HM, Vilela MM, Cunha FG, Abramczuk BM, Reis-Alves SC, Metze K, Lorand-Metze I - BMC Infect. Dis. (2011)

Bottom Line: The non-stimulated, as well as BCG- and PHA-stimulated production of IL2, IL4, IL7, IL10, IL12, IFN-γ and TNF-alpha in mononuclear cell cultures from mothers and infants were quantified using ELISA.Cytokine values in supernatants were similar in infants and their mothers except for IFN-γ and TNF-alpha that were higher in HIV+ mothers, especially in drug abusing ones.A higher maternal age and smoking was associated with a decrease of cord blood CD4+ cells. in uninfected infants born to HIV+ women, several immunological abnormalities were found, related to the residual maternal immune changes induced by the HIV infection and those associated with antiretroviral treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Internal Medicine, Faculty of Medical Sciences, State University of Campinas, Rua Tessalia Vieira de Camargo 126, 13083-887 - Campinas, Brazil.

ABSTRACT

Background: Children born to HIV+ mothers are exposed intra-utero to several drugs and cytokines that can modify the developing immune system, and influence the newborn's immune response to infections and vaccines. We analyzed the relation between the distribution of cord blood lymphocyte subsets and cytokine profile in term newborns of HIV+ mothers using HAART during pregnancy and compared them to normal newborns.

Methods: In a prospective, controlled study, 36 mother-child pairs from HIV+ mothers and 15 HIV-uninfected mothers were studied. Hematological features and cytokine profiles of mothers at 35 weeks of pregnancy were examined. Maternal and cord lymphocyte subsets as well as B-cell maturation in cord blood were analyzed by flow cytometry. The non-stimulated, as well as BCG- and PHA-stimulated production of IL2, IL4, IL7, IL10, IL12, IFN-γ and TNF-alpha in mononuclear cell cultures from mothers and infants were quantified using ELISA.

Results: After one year follow-up none of the exposed infants became seropositive for HIV. An increase in B lymphocytes, especially the CD19/CD5+ ones, was observed in cord blood of HIV-exposed newborns. Children of HIV+ hard drug using mothers had also an increase of immature B-cells. Cord blood mononuclear cells of HIV-exposed newborns produced less IL-4 and IL-7 and more IL-10 and IFN-γ in culture than those of uninfected mothers. Cytokine values in supernatants were similar in infants and their mothers except for IFN-γ and TNF-alpha that were higher in HIV+ mothers, especially in drug abusing ones. Cord blood CD19/CD5+ lymphocytes showed a positive correlation with cord IL-7 and IL-10. A higher maternal age and smoking was associated with a decrease of cord blood CD4+ cells.

Conclusions: in uninfected infants born to HIV+ women, several immunological abnormalities were found, related to the residual maternal immune changes induced by the HIV infection and those associated with antiretroviral treatment. Maternal smoking was associated to changes in cord CD3/CD4 lymphocytes and maternal hard drug abuse was associated with more pronounced changes in the cord B cell line.

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Analysis of T cell proliferation of non-stimulated (A) and PHA-stimulated (B) cord mononuclear cells of a newborn from a HIV-infected mother. After gating of CD3 cells, events were analyzed for size and complexity (forward-scatter and side-scatter gates), from which resting (R1-green) and blast cells (R2-blue) were separated. T-lymphocyte subsets were then analyzed. Dead cells (red) were excluded from all analyses. T cell proliferation was the difference between the percentages of PHA-stimulated blasts and non-stimulated blasts.
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Figure 2: Analysis of T cell proliferation of non-stimulated (A) and PHA-stimulated (B) cord mononuclear cells of a newborn from a HIV-infected mother. After gating of CD3 cells, events were analyzed for size and complexity (forward-scatter and side-scatter gates), from which resting (R1-green) and blast cells (R2-blue) were separated. T-lymphocyte subsets were then analyzed. Dead cells (red) were excluded from all analyses. T cell proliferation was the difference between the percentages of PHA-stimulated blasts and non-stimulated blasts.

Mentions: Cord Blood mononuclear cells (CBMC) were isolated by density gradient centrifugation over Ficoll-Hypaque (Amersham Biosciences, USA), washed, diluted to 1 × 106 cells/mL in RPMI 1640 medium (Sigma, USA), supplemented with 10% human AB serum (Sigma, USA), 1% glutamine (Sigma, USA) and 0.1% gentamycin and stimulated for 6 days with PHA or medium alone at 37°C with 5% CO2 96-well tissue culture plates (NUNC, Denmark). After harvesting in 20 mM EDTA, samples were incubated with human immunoglobulin and then stained with anti-human CD3, CD4 and CD8 (Beckman Coulter, USA) fluorescent antibodies before acquisition and analysis. Only CD3+ T cells were used in the analysis. Resting and blast lymphocytes were gated on the forward and side scatter plot (Figure 2). Dead cells were excluded from all analyses. CD4+ and CD8+ cells were identified in the gate of blast lymphocytes. Proliferation was measured as percentage of CD3+ blasts in the PHA-stimulated well from which basal proliferation without stimulation was subtracted (PHA, Sigma, USA, 7.5 μg/mL).


The impact of maternal HIV infection on cord blood lymphocyte subsets and cytokine profile in exposed non-infected newborns.

Borges-Almeida E, Milanez HM, Vilela MM, Cunha FG, Abramczuk BM, Reis-Alves SC, Metze K, Lorand-Metze I - BMC Infect. Dis. (2011)

Analysis of T cell proliferation of non-stimulated (A) and PHA-stimulated (B) cord mononuclear cells of a newborn from a HIV-infected mother. After gating of CD3 cells, events were analyzed for size and complexity (forward-scatter and side-scatter gates), from which resting (R1-green) and blast cells (R2-blue) were separated. T-lymphocyte subsets were then analyzed. Dead cells (red) were excluded from all analyses. T cell proliferation was the difference between the percentages of PHA-stimulated blasts and non-stimulated blasts.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3040712&req=5

Figure 2: Analysis of T cell proliferation of non-stimulated (A) and PHA-stimulated (B) cord mononuclear cells of a newborn from a HIV-infected mother. After gating of CD3 cells, events were analyzed for size and complexity (forward-scatter and side-scatter gates), from which resting (R1-green) and blast cells (R2-blue) were separated. T-lymphocyte subsets were then analyzed. Dead cells (red) were excluded from all analyses. T cell proliferation was the difference between the percentages of PHA-stimulated blasts and non-stimulated blasts.
Mentions: Cord Blood mononuclear cells (CBMC) were isolated by density gradient centrifugation over Ficoll-Hypaque (Amersham Biosciences, USA), washed, diluted to 1 × 106 cells/mL in RPMI 1640 medium (Sigma, USA), supplemented with 10% human AB serum (Sigma, USA), 1% glutamine (Sigma, USA) and 0.1% gentamycin and stimulated for 6 days with PHA or medium alone at 37°C with 5% CO2 96-well tissue culture plates (NUNC, Denmark). After harvesting in 20 mM EDTA, samples were incubated with human immunoglobulin and then stained with anti-human CD3, CD4 and CD8 (Beckman Coulter, USA) fluorescent antibodies before acquisition and analysis. Only CD3+ T cells were used in the analysis. Resting and blast lymphocytes were gated on the forward and side scatter plot (Figure 2). Dead cells were excluded from all analyses. CD4+ and CD8+ cells were identified in the gate of blast lymphocytes. Proliferation was measured as percentage of CD3+ blasts in the PHA-stimulated well from which basal proliferation without stimulation was subtracted (PHA, Sigma, USA, 7.5 μg/mL).

Bottom Line: The non-stimulated, as well as BCG- and PHA-stimulated production of IL2, IL4, IL7, IL10, IL12, IFN-γ and TNF-alpha in mononuclear cell cultures from mothers and infants were quantified using ELISA.Cytokine values in supernatants were similar in infants and their mothers except for IFN-γ and TNF-alpha that were higher in HIV+ mothers, especially in drug abusing ones.A higher maternal age and smoking was associated with a decrease of cord blood CD4+ cells. in uninfected infants born to HIV+ women, several immunological abnormalities were found, related to the residual maternal immune changes induced by the HIV infection and those associated with antiretroviral treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Internal Medicine, Faculty of Medical Sciences, State University of Campinas, Rua Tessalia Vieira de Camargo 126, 13083-887 - Campinas, Brazil.

ABSTRACT

Background: Children born to HIV+ mothers are exposed intra-utero to several drugs and cytokines that can modify the developing immune system, and influence the newborn's immune response to infections and vaccines. We analyzed the relation between the distribution of cord blood lymphocyte subsets and cytokine profile in term newborns of HIV+ mothers using HAART during pregnancy and compared them to normal newborns.

Methods: In a prospective, controlled study, 36 mother-child pairs from HIV+ mothers and 15 HIV-uninfected mothers were studied. Hematological features and cytokine profiles of mothers at 35 weeks of pregnancy were examined. Maternal and cord lymphocyte subsets as well as B-cell maturation in cord blood were analyzed by flow cytometry. The non-stimulated, as well as BCG- and PHA-stimulated production of IL2, IL4, IL7, IL10, IL12, IFN-γ and TNF-alpha in mononuclear cell cultures from mothers and infants were quantified using ELISA.

Results: After one year follow-up none of the exposed infants became seropositive for HIV. An increase in B lymphocytes, especially the CD19/CD5+ ones, was observed in cord blood of HIV-exposed newborns. Children of HIV+ hard drug using mothers had also an increase of immature B-cells. Cord blood mononuclear cells of HIV-exposed newborns produced less IL-4 and IL-7 and more IL-10 and IFN-γ in culture than those of uninfected mothers. Cytokine values in supernatants were similar in infants and their mothers except for IFN-γ and TNF-alpha that were higher in HIV+ mothers, especially in drug abusing ones. Cord blood CD19/CD5+ lymphocytes showed a positive correlation with cord IL-7 and IL-10. A higher maternal age and smoking was associated with a decrease of cord blood CD4+ cells.

Conclusions: in uninfected infants born to HIV+ women, several immunological abnormalities were found, related to the residual maternal immune changes induced by the HIV infection and those associated with antiretroviral treatment. Maternal smoking was associated to changes in cord CD3/CD4 lymphocytes and maternal hard drug abuse was associated with more pronounced changes in the cord B cell line.

Show MeSH
Related in: MedlinePlus