Urinary Immunoglobulins in Healthy Individuals and Children with Acute Pyelonephritis

Urine samples obtained from children with acute pyelonephritis and from healthy children and adults were analysed with regard to the molecular form and specific antibody activity of urinary immunoglobulins. The urinary IgA and IgG levels were quantified in unconcentrated urine by radioimmunoassay. The children with urinary tract infection had significantly higher levels of IgG and IgA than age-matched controls but not higher than healthy adults. After tenfold concentration, the urine was fractionated on an Ultrogel AcA 22 column, and the IgA, secretory IgA, and IgG in the fractions were determined by radioimmunoassay. IgA in urine from healthy adults was predominantly represented by polymeric IgA linked to secretory component; small quantities of monomeric IgA were also present. IgG eluted in the position of the serum standard. Increased proportions of IgG and monomeric IgA were found in the infected patients. Specific antibody activity of the IgG and IgA classes to antigens of the infecting Escherichia coli strain was detected in whole and in fractionated urine from children with acute pyelonephritis. The specific antibody activity in healthy adults and children was low.     

Isolation of Lymphocytes, Granulocytes and Macrophages

Abstract. This paper presents the standard procedure for isolating lymphocytes and granulocytes from blood, using the Isopaque-Ficoll technique. A procedure for isolating granulocytes and macrophages from peritoneal fluid is also described.     

Sex Differences in the Catecholamine Output of Children

Urinary excretion of catecholamines was studied in 240 normal, healthy children 12 years of age during a passive condition (film exposure) and an active condition (arithmetic test) at school. No sex differences in catecholamine excretion were found during the passive condition, whereas, during the work period, boys excreted significantly more adrenaline and noradrealine than girls. A comparison is made between catecholamine excretion levels of the children and adult subjects examined in other studies.     

Differences in Death Concepts Between Children Wither Cancer and Physically Healthy Children

The objectives of the current study were (a) to investigate differences in concepts of death between healthy children and children with cancer and (b) to investigate within the oncology sample the effects of disease,related variables and previous experience with death on concepts of death. Study participants included 32 children with cancer (ages = 3 to 16 years) and 32 healthy children matched on age. Analyses of a structured interview indicated that healthy children were more likely to exhibit the concepts of immanent justice and personal death than children with cancer. Point-biserial correlations conducted within the oncology group indicated that 3- to 6-year-old children who had had personal experiences with death (loss of a loved one) were more likely to exhibit the concepts of universality, personal death, and irrevocability. Implications for clinical practice are reviewed, and directions for future research are discussed.     

Functional and Phenotypic Changes in Circulating Lymphocytes from Hospitalized Zambian Children with Measles

Measles is associated with immunosuppression and increased susceptibility to secondary infections and is a particular problem in developing countries. Lymphocyte changes accompanying immune activation and regulation of the immune response may contribute to immunosuppression. To evaluate lymphocyte changes during measles, children (n = 274) hospitalized with measles in Lusaka, Zambia, were evaluated at entry, discharge, and 1-month follow-up and compared to healthy Zambian children (n = 98). Lymphopenia was present on hospital admission and reflected decreased CD4 and CD8 T cells but resolved quickly. Lymphopenia was most marked in girls, in those with temperatures of >38.5°C, and in malnourished children. CD4/CD8 ratios were decreased at all time points and were lower in boys than in girls at discharge and follow-up. Spontaneous death occurred in cultured lymphocytes, and the proportions of freshly isolated cells undergoing apoptosis, based on annexin V and propidium iodide staining, were increased. Surface Fas was increased on both CD4 and CD8 T cells compared to controls, and expression was greater on CD4 T cells and was inversely correlated with lymphocyte viability in culture at study entry. Mitogen stimulation of lymphocytes improved viability, but inhibitors of Fas, tumor necrosis factor (TNF)-related apoptosis-inducing ligand, and TNF did not. Plasma levels of β₂ microglobulin and soluble Fas, Fas ligand, CD8, CD4, and TNF receptor were increased, and soluble CD8 was higher in boys than in girls. The multiple effects of measles on lymphocytes from Zambian children include decreased numbers in circulation, increased activation, and increased susceptibility to cell death, with substantive differences in the magnitude of these changes between boys and girls.     

Differences in Circulating Dendritic Cell Subtypes in Pregnant Women, Cord Blood and Healthy Adult Women

Different subtypes of dendritic cells (DC) influence the differentiation of naíve T lymphocytes into T helper type 1 (Th1) and Th2 effector cells. We evaluated the percentages of DC subtypes in peripheral blood from pregnant women (maternal blood) and their cord blood compared to the peripheral blood of healthy non pregnant women (control). Circulating DC were identified by flow cytometry as lineage (CD3, CD14, CD16, CD19, CD20, and CD56)-negative and HLA-DR-positive cells. Subtypes of DC were further characterized as myeloid DC (CD11c⁺/CD123^±), lymphoid DC (CD11c^-/CD123⁺⁺⁺) and less differentiated DC (CD11c^-/CD123^±). The frequency of DC out of all nucleated cells was significantly lower in maternal blood than in control (P<0.001). The ratio of myeloid DC/lymphoid DC was significantly higher in maternal blood than in control (P<0.01). HLA-DR expressions of myeloid DC as mean fluorescence intensity (MFI) were significantly less in maternal blood and in cord blood than in control (P<0.001, respectively). The DC differentiation factors, TNF-α and GM-CSF, released from mononuclear cells after lipopolysaccharide stimulation were significantly lower in maternal blood than in control (P<0.01). The distribution of DC subtypes was different in maternal and cord blood from those of non-pregnant women. Their role during pregnancy remains to be determined.     

Surface Immunoglobulins of Circulating Lymphocytes in Mouse Plasmacytoma III. The Effect of Plasmacytoma RNA on the Immune Response

RNA extracted from several plasmacytomas of BALB/c mice was previously shown in in vitro and in vivo experiments to induce the synthesis of receptors on B cells with The immunochemical characteristics of the respective plasmacytoma globulin. These findings were also produced with immuno-globulin-free RNA extracted from the plasma of plasmacytoma animals. The results of the present study indicate that the injection of 0.5-1 mg of RNA into CF₁ mice also has an inhibiting effect on the primary response to sheep erythrocytes (SRBC) as measured by agglutinin titers and plaque forming cells. On the other hand, RNA injected prior to the second injection of SRBC did not inhibit the secondary response. RNA from normal tissue and RNase-inactivated RNA were ineffective. These findings suggest that an RNA fraction of the plasmacytoma participates in the mediation of the characteristic impairment of the primary immune response in myeloma.     

Racial Differences in Personality, Behavior, and Family Environment in Minneapolis School Children

Racial differences in personality, behavior, and family environment of lower elementary school children were examined in a sample of 433 black and 897 white children. Numerous significant differences in scores on scales of the Missouri Children''s Picture Series, the Missouri Children''s Behavior Checklist, and the Family Environment Scale persisted after adjustment for socioeconomic status.     

Differences between Metabolically Healthy vs Unhealthy Obese Children and Adolescents

Obesity is on the rise worldwide. An obesity subtype, metabolically healthy obese (MHO), is resilient to unfavorable metabolic and cardiovascular effects. Factors predicting MHO phenotype are not well characterized. We aimed to identify MHO and metabolically unhealthy obese (MUO) children and adolescents with respect to metabolic factors, and to find predictors of MHO subtype. A retrospective chart review was done on children, ages 4–19 years, 99% African–American/Caribbean, with BMI ≥95th %tile. MUO was defined as meeting ≥1 of the following: fasting glucose ≥100 mg/dl, HbA1c >5.6%, BP ≥90th %tile, TG ≥150 mg/dl, or HDL <40 mg/dl. Study included 189 subjects, 37.6% were MHO and 62.4% MUO. MHO subjects were younger (mean ± SD, 11.6 ± 3.3 vs 12.9 ± 3.2 years; p < 0.009) and had lower BMI %tile (98.4 ± 1.4 vs 98.8 ± 2.1; p < 0.04), smaller waist (94.2 ± 15.2 vs 101.4 ± 17 cm; p < 0.003) and hip circumferences (105.3 ± 15.6 vs 113.5 ± 15.4 cm; p < 0.001), lower fasting insulin (18.5 ± 10.2 vs 24.2 ± 14.3 μU/ml; p < 0.022), and lower HOMA-IR (4.1 ± 2.4 vs 5.5 ± 3.6; p < 0.022). Acanthosis nigricans was noted less frequently in MHO than MUO (p < 0.005). In stepwise logistic regression, age and BMI %tile were significant predictors of MHO. We found that 38% of obese children are MHO. They are younger and have lower BMI %tiles. Lifestyle modification initiated at an early age may prevent metabolic abnormalities.     

Interaction of Macrophages and Lymphocytes in Rat Skin Allografts

Light microscopy of 2μm sections of rejecting rat skin allografts, embedded in hydroxyethyl methacrylate, revealed among the cells infiltrating the graft base extravascular macrophages containing a small lymphocyte. Toluidine blue staining indicated DNA degradation in some of these phagocytosed lymphocytes. More frequently small lymphocytes were in intimate contact with the surface of the macrophages, resembling 'Periopolesis', which others have previously observed in vitro. These macrophage-lymphocyte interactions were not seen in sections of autografts. Despite a previous report that diphenylhydantoin (phenytoin) impairs macrophage function, these macrophage-lymphocyte interactions were present in grafts placed in rats receiving this drug. This treatment did not hasten or delay the onset of graft rejection. These in vivo findings both accord with recent in vitro studies on the mechanisms of phagocytosis and with reports that phagocytosis is one of the effector mechanisms in allograft rejection. However, macrophage phagocytosis of lymphocytes has also been observed in testicular lymph collected from conscious normal sheep.     

Production and Secretion of Immunoglobulins by in Vitro-Activated Human B Lymphocytes

Activation induced by pokeweed mitogen in cultures of mononuclear cells from human blood was followed sequentially by simultaneous quantitation of live cells, thymidine incorporation, cells displaying cytoplasmic IgM, IgG, IgA or IgD, cells secreting IgM, IgG or IgA and cumulated IgM secretion. Maximal cellular activity was found after 7 days of culture, with means of 16000 IgM-, 20700 IgG- and 9900 IgA-secreting cells per 10⁶ originally cultured cells. The cumulated IgM secretion after 21 days of culture averaged 10400 ng per 10⁶ originally cultured cells. A close correlation was found between the number of IgM-secreting cells and the cumulated IgM secretion.     

Interaction of Macrophages and Lymphocytes in Rat Skin Allografts

Light microscopy of 2μm sections of rejecting rat skin allografts, embedded in hydroxyethyl methacrylate, revealed among the cells infiltrating the graft base extravascular macrophages containing a small lymphocyte. Toluidine blue staining indicated DNA degradation in some of these phagocytosed lymphocytes. More frequently small lymphocytes were in intimate contact with the surface of the macrophages, resembling ‘Periopolesis’, which others have previously observed in vitro. These macrophage-lymphocyte interactions were not seen in sections of autografts. Despite a previous report that diphenylhydantoin (phenytoin) impairs macrophage function, these macrophage-lymphocyte interactions were present in grafts placed in rats receiving this drug. This treatment did not hasten or delay the onset of graft rejection. These in vivo findings both accord with recent in vitro studies on the mechanisms of phagocytosis and with reports that phagocytosis is one of the effector mechanisms in allograft rejection. However, macrophage phagocytosis of lymphocytes has also been observed in testicular lymph collected from conscious normal sheep.     

Stimulation of Human Lymphocytes by Allogenic Macrophages in vitro

Macrophages were obtained from human peripheral blood by incubating mononuclear cells in plastic tissue culture flasks. After 1-14 days, the cells were used in mixed lymphocyte macrophage cultures. Macrophages could not, themselves, be stimulated to proliferation by allogenic cells, but stimulated allogenic lymphocytes. By the use of responding cells and stimulating macrophages from HLA-D homozygous individuals, the HLA-D determinants could be shown to be responsible for the stimulation of allogenic lymphocytes.     

Role for Macrophages and Thymus-Derived Lymphocytes in Cholera Toxin-Induced Immunosuppression

The exo-enterotoxin derived from Vibrio cholerae bacilli has marked immunomodulating activities, both in vivo and in vitro. In the present study, the mechanism whereby cholera toxin depresses the antibody-forming ability of murine splenocytes was investigated by in vitro reconstitution experiments. Spleen cells derived from mice treated with cholera toxin 2 days earlier were markedly deficient in their ability to respond to sheep erythrocytes upon challenge immunization in vitro. Addition of graded numbers of normal spleen cells to spleen cell cultures from toxin-treated mice partially restored the antibody response. Adherent splenocyte populations were even more effective in restoring antibody formation. Normal peritoneal exudate cells rich in macrophages were also capable of restoring the antibody-forming ability of toxin-pretreated splenocytes. Furthermore, thymus (T)-derived spleen cells from normal mice, as well as sheep erythrocyte "educated" T cells, were capable of restoring antibody formation to normal levels. The importance of T lymphocytes in restoring immune competence of spleen cell cultures from toxin-treated mice was shown by additional experiments in which T-depleted cell preparations were found to be ineffective in restoring antibody activity. These studies point to macrophages and T-derived lymphocytes as a major target for cholera toxin-induced immunosuppression.     

Normal Value of Immunoglobulins IgA,IgG, and IgM in Iranian Healthy Adults,Measured by Nephelometry

Abstract

Local reference ranges of immunoglobulins are required for studies and clinical interpretation. In this study, serum levels of IgA, IgG, and IgM in 914 Iranian healthy adult blood donor volunteers, aged 18–55 years, were measured by nephelometry. Our data showed that serum reference intervals of IgA, IgG, and IgM in subjects were 72–375, 636–1518, and 39–283 mg/dL, respectively. Data analysis showed a significant difference between the male and female subjects only for IgM; thus, the means of IgM in females were higher than for males (p<0.05). Correlation coefficient r for paired samples showed no statistically significant relationship between age and each Ig (p>0.05). Comparison of this study with others demonstrated that results are similar; some differences are probably related to ethnic differences. Therefore, our results can be considered as a source of reliable local reference for use in laboratories.     

Age,Sex, and Individual Differences in Children's Defensive Styles

Defensiveness in 4- and 6-year-old boys and girls was measured using a social desirability scale, a parent rating form of children's defensive behavior, and a semiprojective emotional attribution story-telling task. On the emotional attribution task, boys attributed less intense emotions to stories than did girls, and older children attributed less intense emotions to stories than did younger children. The quality and intensity of children's emotional attributions were related to their defensive styles, and individual differences in children's defensive styles occurred as early as age 4.     

The Variability of Immunoglobulins and Albumin in Salivary Secretions of Children

The variation of concentrations of immunoglobulins and albumin in consecutive daily collections of saliva was studied in 33 infants, aged 6 months to 5 years, for periods ranging from 16 to 26 days. The concentration and the within-child variability of IgA and albumin and the detection of IgG and IgM in saliva increased with age. Between-child variances were greater than the within-child variances by a factor of 2.8 for log (IgA) and 1.3 for log (albumin). The geometric mean IgA levels were consistently higher and IgG was detected more frequently during upper respiratory tract infections compared with periods of non-infection. There were no changes in albumin levels between infection and non-infection periods, suggesting a local immune response rather than serum leakage. There were significant within-child correlations (autocorrelations) between levels of IgA in saliva collected on consecutive days and samples collected up to 3 days apart. The autocorrelations between levels of albumin were significant for samples collected up to 2 days apart. The autocorrelation for IgA was significantly greater during infection periods compared with non-infection periods for samples collected on consecutive days.