Human colostrum T lymphocytes and their effector cytokines actively aid the development of the newborn immune system.

Colostrum contains soluble and cellular components, the latter mainly T lymphocytes. We expanded in vitro colostrum T lymphocytes (CoTL) to evaluate phenotype and capability of cytokine production. We also considered paired cord blood T-lymphocytes (CBTL) representing the newborn "virgin" immune system. CoTL showed memory phenotype while CBTL expressed mainly naïve phenotype. CoTL included a balanced percentage of helper and cytotoxic subsets. We observed higher percentages of IL-2 (p=0.003) and IL-4 (p=0.027) producing cells by helper rather than by cytotoxic T lymphocytes. The greatest percentage of IFN-gamma producing cells was in cytotoxic cells (p=0.0048), while no difference was found for IL-10. Cord blood samples consisted of a statistically significant greater percentage of helper than cytotoxic cells (p<0.001), with a low percentage of cytokine producing cells, confirming the immaturity of the newborns immune system. CBTL percentage of IL-2 producing cells was higher for helper than cytotoxic subset (p<0.001). We observed a greater percentage of IFN-gamma (p=0.001), IL-4 (p=0.003) and IL-10 (p<0.001) producing cells by cytotoxic than helper T lymphocytes. CoTL demonstrated to protect the newborn through the mothers previous immune experience and to supply active cytokines, which can help the postnatal development of both T type 1/T type 2 response.

Escherichia coli specific secretory IgA and cytokines in human milk from mothers of different ethnic groups resident in northern Italy.

Breast milk supplies many bioactive components. Neonates protection from pathogenic bacteria is mainly attributable to secretory IgA antibodies present in human milk in an amount depending on previous antigenic exposure. To bring new details into the field of immunological memory in secretory immunity, we evaluated the production of s-IgA specific for E. coli (E. coli s-IgA), and of pro-inflammatory (IL-6 and IL-8) or anti-inflammatory (IL-10) cytokines in the milk of mothers of different ethnic groups exposed in the past to poor conditions, but nowadays living in Italy in adequate conditions. Mothers from Italy, Africa, Asia and Eastern European Countries were included in the study. Anti-E. coli s-IgA, IL-6, IL-8 and IL-10 were determined by ELISA. Breast milk of all the foreign mothers presented higher levels of E. coli s-IgA than Italians, and for Asian and African mothers were significative (p=0.031 and p=0.015, respectively). Milk from women of Eastern European Countries revealed the highest IL-8 levels (p=0.026), while milk from Asian women presented the greatest concentration of IL-6 (p=0.04); however, the Africans reported the lowest concentrations of IL-10 (p=0.045). Since all the mothers had been living in Italy for some time, we believe that the presence of high levels of E.coli s-IgA, supported by high levels of pro-inflammatory cytokine, is part of a persisting immunological secretory memory.

IFN-gamma low production capacity in type 1 diabetes mellitus patients at onset of disease.

In type 1 diabetes mellitus (T1DM), cytokines can be directly cytotoxic to beta-cells, and/or play an indirect role influencing some cells of the immune system. Since several factors could impair cytokine serum levels, the purpose of our study was to longitudinally evaluate intracellular cytokines, in T1DM patients, and in subject at risk, by flow cytometry analysis. At T1DM onset we observed significantly lower percentage of peripheral CD4 + and CD8 + cells producing IFN-gamma in patients compared to controls and subjects at risk. The 15-month follow-up patients showed significantly lower percentage of CD4 + and CD8 + cells producing IFN-gamma compared to the other groups. At 8-year follow-up no significant differences were observed among the groups in the percentage of cells producing cytokines. We could have considered "exhausted cells" or these T cell subsets may be migrated from peripheral blood to pancreas. On the other hand, our results are in agreement with those reported in literature: in animal model the absence of IFN-gamma production makes beta-cells highly susceptible to viral infection and subsequent attack by natural killer cells, which lead to hyperglycaemia and diabetes mellitus.

Neonatal outcome in patients with rheumatic disease.

Rheumatic autoimmune diseases have a higher prevalence in women, particularly during their childbearing age. Due to improved management, an increasing number of patients plan and carry out one or more pregnancies. Therefore, a growing interest is being paid to the possible consequences of maternal disease and associated treatment on the fetus and newborn infant. If maternal disease is characterized by the presence of IgG isotype autoantibodies, these can cross the placenta with possible antibody-mediated damage to the fetus. This is typically the case of the so called neonatal lupus erythematosus (NLE); a similar mechanism has been shown in infants of patients with immune thrombocytopenic purpura (ITP) and, less frequently, in those from mothers with antiphospholipid syndrome (APS). Indeed, this last condition is often responsible for placental, rather than neonatal, pathology. In addition, immunosuppressive and other drugs administered to the mothers during pregnancy and lactation might affect the fetal and neonatal immune system development. Finally, mothers disease and/or treatment could be related to neuropsychological alteration reported in some of their children.

Antigen-specific T cell response in infants after recombinant hepatitis B virus vaccination at birth: evaluation of T helper lymphocyte diversity.

Recombinant hepatitis B virus antigen (rHBsAg)-specific CD4+ T cell clones (TCC) were isolated and expanded from the peripheral blood of nine children vaccinated at birth against the hepatitis B (HB) virus. Four of them responded with protective antibody production (responders), three subjects were unable to produce detectable antibody levels even after revaccination (nonresponders), and two infants produced antibodies only after revaccination (slow responders). TCC were then characterized for their ability to produce cytokines known to be important for T cell expansion (interleukin-2, IL-2) and/or effector functions (IL-4, IFN-gamma, IL-10). Results demonstrated that the frequency of rHBsAg-specific TCC in the samples of nonresponders was comparable to or higher than that in the samples of responders. Nevertheless, the majority of TCC obtained from responders or from slow responders before revaccination displayed the T helper 1 (T(H1))-dominant phenotype, while the majority of TCC obtained from nonresponders were nonpolarized T lymphocytes. After revaccination, the distribution of the different T(H) subsets in slow responders was heterogeneous. Overall, our present data suggest that an absence or delay in developing an rHBsAg-specific antibody response to vaccination is not associated with the capacity to generate an Ag-specific T cell response. However, compared to responders, nonresponding infants react to the rHBsAg vaccination with a reduced capacity to expand and differentiate toward polarized T(H) cells.

Effect of general and epidural anaesthesia on thyroid hormones and immunity in neonates.

BACKGROUND: The aim of this study was to verify if variations of thyroid hormones related to circumstances of delivery and mode of maternal anaesthesia can contribute to neonatal neutrophil respiratory burst and natural killer cell activity. METHODS: We evaluated 10 infants born by vaginal delivery (group A), 10 infants born by caesarean section after epidural anaesthesia with lidocaine (group B) and 10 infants born by caesarean section after general anaesthesia with sevoflurane (group C). RESULTS: A significant reduction of neutrophil respiratory burst test was found in groups A and C compared with group B. Natural killer cell (NK) activity with an effector : target ratio of 30 : 1 (NK30) and 10 : 1 (NK10) was significantly higher in group A compared with the B and C groups. In addition, thyroid stimulating hormone (TSH) concentration was significantly reduced in group A compared with the B and C groups. A significant negative correlation was found between TSH and NK30 or NK10. CONCLUSIONS: Our results suggest that the mode of delivery and anaesthesia can significantly modify the endocrine-immune system in the newborn. Caesarean section delivery with regional anaesthesia seems to produce fewer modifications of neonatal immune function compared with general anaesthesia.

Increment of recombinant hepatitis B surface antigen-specific T-cell precursors after revaccination of slow responder children.

The aim of the study was to investigate the in vitro T-cell response to recombinant hepatitis B (rHBsAg) in a group of children (defined as "slow responders") vaccinated at birth, presenting antibody levels < 10 mIU/ml after the vaccination schedule, and developing anti-rHBs antibodies after revaccination. T-cell mediated immune response towards rHBsAg was evaluated in 35 healthy children in "bulk" culture experiments (19 responders and 16 slow responders) and by limiting dilution analysis (nine responders and five slow responders) to quantify the frequency of proliferating T lymphocyte-precursors (PTL-p). Before the booster dose, lymphocytes from slow responder children failed to proliferate to rHBsAg, while a normal proliferation was observed in all responders. A statistically significant difference in rHBsAg-specific PTLp frequencies was observed between the two groups. Among the slow responder group, a significant increase of PTLp was observed after the supplementary vaccine dose.Nevertheless, PTLp frequencies remained significantly lower than those measured in responders. These results suggest a role for follow-up of slow responder children over time, in order to perform booster vaccination when inadequate anti-HBs titre is present.

Immunophenotypic changes of fetal cord blood hematopoietic progenitor cells during gestation.

We measured cell surface expression of CD34, HLA-DR, CD38, CD19, CD33, CD71, and CD45 antigens in the hematopoietic progenitor cells of fetal cord blood to investigate immunophenotypic changes at different gestational ages. These antigens were identified by flow cytometry in 11 fetuses (gestational age 19-24 wk, in 12 preterm (25-28 wk) and in ten newborn infants born at term. The frequency and number of CD34+ cells were higher in the blood of the 11 fetuses; in addition, a statistically significant inverse correlation between number of CD34+ cells and advancing gestational age was noted. The numbers of CD34+ CD19+, CD34+ CD33+, and CD34+ CD45+ coexpressing cells were significantly higher in the fetuses, whereas CD34+ CD38+ cells were more represented in the neonates at term. Gestational age was inversely correlated with the number of CD34+ CD19+ and CD34+ CD33+ coexpressing cells. A positive correlation between gestational age and CD34+ CD38+ cells was noted. The number of CD34- CD19+, CD34- CD38+, and CD34- CD45+ cells was higher in term infants; furthermore, a significant correlation between advancing gestational age and CD34- CD38+ or CD34- CD45+ cells was demonstrated. The proliferative capacity was also higher at lower gestational ages. These data suggest that the development and lineage commitment of fetal cord blood hematopoietic progenitor cells are very active during the last two trimesters of pregnancy. The most significant changes of hematopoietic cells maturation seem to occur within 25 wk of gestation.

Leukocyte counts in relation to the method of delivery during the first five days of life.

The goal of the present study was to evaluate total and differential leukocyte counts during the first 5 days of life in relation to the method of delivery. We included 203 healthy term infants; of these, 114 were born by vaginal delivery, and 89 by elective cesarean section. Total and differential leukocyte counts were evaluated at the following intervals: 0-6, 7-12, 13-24, 25-48, 49-72, 73-96, and 97-120 h after birth. The cord serum cortisol level was measured as an indicator of the degree of delivery-related stress. Mean leukocyte and neutrophil counts were higher in infants born by vaginal delivery in cord blood and up to 12 h of life. No significant differences were observed in the immature: total neutrophil ratios between the two groups of infants. The cord serum cortisol level was higher in vaginally delivered infants. A significant correlation was found between cortisol and leukocyte, neutrophil, or lymphocyte counts. The method of delivery produces significantly different total leukocyte and neutrophil counts during the first 12 h after birth; after this time, there appears to be no more variation of leukocyte counts during the first 5 days of life.

Effect of lidocaine on neutrophil chemotaxis in newborn infants.

Anesthetic drugs can influence the immune system, particularly granulocyte function. The goal of the present study was to evaluate if lidocaine used for epidural anesthesia during cesarean section can influence neonatal neutrophil chemotaxis. We measured chemotaxis and plasma cord lidocaine and cortisol levels in (A) 15 infants born by cesarean section with epidural anesthesia, (B) 15 infants born by vaginal delivery, and (C) 20 infants born by cesarean section with general anesthesia. Chemotaxis levels were significantly lower in group A infants (35.5 +/- 16.1 microns) compared to groups B (54.6 +/- 10.5 microns) and C (71.4 +/- 23 microns). The highest cortisol levels were observed in vaginally delivered infants. A significant inverse relationship was observed between chemotaxis and lidocaine levels (r = -0.6, P = 0.016) in infants born by cesarean section after epidural anesthesia, while no significant correlation was observed between chemotaxis and cortisol level. In conclusion, lidocaine, transferred through the placenta to the fetus during epidural anesthesia, may have an inhibitory effect on chemotaxis.

Serum concentration of granulocyte colony stimulating factor in term and preterm infants.

UNLABELLED: We measured serum granulocyte colony stimulating factor (GCSF) concentration and absolute neutrophil count in four groups of infants: (1) 15 healthy term newborn infants; (2) 21 healthy preterm newborn infants, with mean (SD) birth weight 1583 (533) g, and gestational age 32.0 (3.8) weeks; (3) 5 infected newborn infants; (4) 22 6-month-old control infants. Median (range) serum GCSF concentration was 132.2 (41.5-176.0) pg/ml in term infants, 51.5 (1.8-175.7) pg/ml in preterm infants and 138.9 (54.1-449.8) pg/ml in 6-month-old control infants, with a significant reduction in preterm infants, as compared to term and control infants. GCSF levels were significantly higher in the infected infants, as compared to healthy neonates. CONCLUSION: A significant positive relationship was found in term and preterm infants between serum GCSF concentration and gestational age or birth weight. No relationship was found between serum GCSF concentration and neutrophil count. The low GCSF baseline levels may contribute to the increased incidence and severity of infection in preterm infants.

Immunogenicity and antigenicity of a partially hydrolyzed cow's milk infant formula.

We evaluated the immunogenicity and antigenicity of a formula based on partially hydrolyzed cow's milk whey protein in infants at risk of atopy and in controls. Total IgE and specific IgE, IgG, and IgG4 subclass antibodies against egg albumin and cow's milk alpha-lactalbumin, casein, and beta-lactoglobulin were measured by radioimmunoassay of cord blood and of peripheral blood at 5 days and 6 months of life in five groups of infants: 16 breast-fed infants at risk of atopy (group 1), 21 partially hydrolyzed whey formula-fed infants at risk of atopy (group 2), 14 formula-fed infants at risk of atopy (group 3), 10 breast-fed control infants (group 4), and 13 formula-fed control infants (group 5). Total IgE concentration was significantly lower in group 2 at 6 months than in groups 3 and 5 infants and similar to that observed in groups 1 and 4 infants. The concentration of specific antiegg and anti-cow's milk protein IgG and of specific anti-cow's milk alpha-lactalbumin and beta-lactoglobulin IgG4 subclass antibodies was significantly reduced in group 2 as compared to group 3 infants and similar to that found in breast-fed infants. In conclusion, the partially hydrolyzed formula was less immunogenic and antigenic than a traditional formula and was as immunogenic and antigenic as breast milk.

[Bioavailability of a delayed-action aminophylline preparation]. / Biodisponibilità di un preparato ritardo di aminofillina.

Absorption of theophylline from one commercial product labelled as aminophylline sustained release was compared to the absorption from an oral solution of aminophylline in a single-dose bioavailability study. Aminomal-R tablets had bioavailability (101.2 +/- 19) statistically indistinguishable from that of the standard but showed significantly slower absorption (peak times of 3.6 +/- 1.1 h vs 1.3 +/- 0.8 h) and lower peak plasma concentrations (16.8 +/- 4.7 mg/l/1 g aminoph. dose vs 21.1 +/- 4.2 mg/l/1 g aminoph. dose). Projections of plasma concentrations upon multiple dosing were made from single dose data: the dosage interval (every 12 h) concentration ratio which reflects both the frequency of dosing and the entry of the drug into and removal from the body was of 1.8 vs 3.1.

An enzyme-histochemical approach to the study of the human bone-marrow stroma.

The different cells and structures of the bone-marrow stroma were studied with histochemical markers. Reticulum cells, endothelial cells and macrophages were identified by their morphologic characteristics as well as by their different positivity to alkaline-phosphatase, alpha-naphthyl-acetate-esterase and Prussian blue staining. Similarities, as well as distinguishing features of all these cells, possibly related to their common or different origin and functional properties, are described.

The clinical pharmacology of drug interactions.

Preliminary evaluation of pre-established and extemporaneous combinations showed that any combination requires broad, precise, and scientifically valid documentation. Every drug has to be evaluated according to physical, chemical, and pharmacologic points of view, either individually or in combination, and above all be adapted to the individual needs of each patient. To make a realistic contribution to evaluating and devising new combinations, specialized centers should be instituted such as the Central Admixture Service of S. Matteo Hospital in Pavia.