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Biomedical subjects

M E Rossi

Publications and source records attributed to M E Rossi.

At least 55 records · Page 3Linked to original sources

Immune interferon enhances functional properties of human granulocytes: role of Fc receptors and effect of lymphotoxin, tumor necrosis factor, and granulocyte-macrophage colony-stimulating factor.

We report here a comparative study of the effects of several cytokines known to affect myeloid cell differentiation on functional properties of human mature granulocytes. We show that recombinant interferon-gamma (rIFN-gamma), recombinant granulocyte/macrophage-colony stimulating factor (rGM-CSF), recombinant tumor necrosis factor (rTNF), and lymphotoxin (LT) purified to homogeneity are potent stimulators of polymorphonuclear cells (PMN) activity. All cytokines enhance antibody-dependent cell-mediated cytotoxicity (Ab-CMC) mediated by human PMN; however, rGM-CSF, rTNF, and LT have an immediate and short-lived effect on the PMN, whereas the activation by rIFN-gamma requires several hours of induction but can be observed up to 24 to 48 hr of culture. Only the effect of rIFN-gamma is in part dependent on induction of a high-affinity FcR for monomeric IgG on PMN, as suggested by two-color sorting analysis, and on mechanisms that result in prolonged survival of PMN in a functionally active state to mediate oxidative burst, phagocytosis, and bactericidal activity. Greater enhancement of Ab-CMC is obtained by using rIFN-gamma in combination with the other cytokines. Our data indicate that cytokines previously defined on the basis of their cytotoxic effects mediate a wide spectrum of activities on mature myeloid cells and provide evidence for their possible role in vivo, alone or in combination with rIFN-gamma, in modulating functional activities of cells responsible for non-adaptive systems of defense.

Antibody-Dependent Cell Cytotoxicity↗

Determination of monoenoic fatty acid double bond position by permanganate-periodate oxidation followed by high-performance liquid chromatography of carboxylic acid phenacyl esters.

This investigation was carried out to develop methods for a reverse-phase, high-performance liquid chromatography analysis of the monocarboxylic and dicarboxylic acids produced by permanganate-periodate oxidation of monoenoic fatty acids. Oxidation reactions were performed using [U-14C]oleic acid and [U-14C]oleic acid methyl ester in order to measure reaction yields and product distributions. The 14C-labeled oxidation products consisted of nearly equal amounts of monocarboxylic and dicarboxylic acid (or dicarboxylic acid monomethyl ester), with few side products (yield greater than 98%). Conversion of the carboxylic acids to phenacyl esters proceeded to completion. HPLC of carboxylic acid phenacyl esters was performed using a C18 column with a linear solvent gradient beginning with acetonitrile/water (1/1) and ending with 100% acetonitrile. Excellent resolution was achieved for all components of a mixture of C5 through C12 monocarboxylic acid phenacyl esters and C6 through C11 dicarboxylic acid phenacyl esters. Resolution was also achieved for all components of a mixture of C5 through C12 monocarboxylic acid phenacyl esters and C6 through C11 dicarboxylic acid monomethyl, monophenacyl esters. The resolution obtained by HPLC demonstrates that, for a wide range of monoenoic fatty acids, both products of a permanganate-periodate oxidation can be identified on a single chromatogram. Free fatty acids and fatty acid methyl esters were analyzed with equal success. Neither the oxidation nor the esterification reaction caused detectable hydrolysis of methyl ester. The method is illustrated for free acids and methyl esters of 14:1 (cis-9), 16:1 (cis-9), 18:1 (cis-6), 18:1 (cis-9), and 18:1 (cis-11).

Acetophenones↗

Association between morphologic distortion of sickle cells and deoxygenation-induced cation permeability increase.

We hypothesized that the deoxygenation-induced increase in cation permeability of sickle cells was related to mechanical distention of the membrane by growing HbS polymer within the cell. To test this hypothesis, we determined the effect of deoxygenation on cation fluxes in sickle cells under conditions that restricted or permitted extensive growth of polymer, producing different degrees of membrane distention. Manipulation of suspending medium osmolality for density-isolated high and low mean cell hemoglobin concentration (MCHC) cells was used to regulate the extensional growth of polymer bundles and hence membrane distortion. For initially low MCHC cells, the deoxygenation-induced increase in both Na and K fluxes was markedly suppressed when the MCHC was increased by increasing the osmolality. This suppression corresponded to the inhibition of extensive morphologic cellular distortion. For initially high MCHC, ISC-rich cells, deoxygenation had minimal effect on K permeability. However, reduction of MCHC by a decrease in osmolality produced a concomitant increase in cation permeability and cellular distortion. These observations support the idea that the sickling-associated increase in membrane permeability is related to mechanical stress imposed on the membrane by bundles of HbS polymer.

Anemia, Sickle Cell↗

The multitransfused beta-thalassemic child: a model for the study of IgE response.

Positive skin tests to common inhalant allergens and the frequency of respiratory atopic diseases were determined in a population of multitransfused beta-thalassemic patients and in their parents and siblings. Positive skin test responses and respiratory atopic diseases were significantly less frequent in patients than in their relatives and in controls. Negative patients differed from positive patients as to the age at first transfusion, since the former had started transfusion earlier than the latter. It is speculated that in beta-thalassemic patients, allergen-specific IgG acquired early in life through transfusions suppresses the synthesis of allergen-specific IgE.

Adolescent↗

Altered T cell subsets and function in polytransfused beta-thalassemia patients: correlation with sex and age at first transfusion.

Monoclonal antibodies (OKT series) have been used to investigate possible modifications of T lymphocytes and T lymphocyte subsets in 65 multiply transfused beta-thalassemia patients. No significant difference was observed in percentage and absolute number of OKT3-, OKT4-, and OKT8-positive cells when compared to controls. A subgroup of patients (10 patients, 15.3%), however, could be selected who showed a reversal of OKT4/OKT8 ratio. These patients did not differ from the others as to age, number of transfusions, frequency of splenectomy, ferritin levels, hepatitis B markers, chronic liver disease incidence, and numbers of B lymphocytes and natural killer cells. The features distinguishing this group from the remaining patients were: decreased mitogen responsiveness; early age when first transfused; high incidence of males (90%). Immunological investigation was done in 2 occasions, 1 year apart, but no significant modification was observed in these patients. These findings suggest that in beta-thalassemia patients transfusion therapy started very early in life may be responsible for persistent immunological modifications. The susceptibility to such modifications might be greater in males.

Adolescent↗

Should hepatitis B surface antigen positive mothers breast feed?

Breast fed infants may be at greater risk of mother to infant hepatitis B virus infection compared with formula fed infants. We studied 85 infants born to 84 hepatitis B surface antigen positive mothers (only two of whom were hepatitis B e positive), and who had received immunisation against hepatitis B virus. Our results indicate that breast feeding does not increase the risk of developing hepatitis B virus infection in infants born to these mothers if immunisation is carried out.

Breast Feeding↗

Interference of hepatitis B virus surface antigen with natural killer cell function.

The influence of purified hepatitis B virus surface antigen (HBsAg) preparations or of supernatants derived from PLC/PRF/5 cell line (which produces HBsAg) on human natural killer (NK) activity was examined. Lymphocytes pre-incubated with HBsAg and subsequently washed showed a significant decrease in NK cytotoxicity against K-562 target cells. This effect was reversible and dose-dependent. In addition, pre-incubation with either HBsAg or PLC/PRF/5 supernatants inhibited in a reversible manner lymphocyte--K-562 conjugates and the binding of B73.1 monoclonal antibody (MoAb), which recognizes Fc receptors on NK cells. This effect was not observed with HNK-1, T3, T4, T6, T8 and T11 MoAb. HBsAg was non-toxic to lymphocytes, and ineffective with K-562 target cells. Beta-interferon did not modify HBsAg-mediated inhibition, when added either before or during the contact with HBsAg. Moreover, no modification was observed when neutrophils (at various neutrophil:lymphocyte ratios) were added, even though HBsAg is known to stimulate neutrophils to produce oxygen radicals which may modulate NK activity. We speculate that HBsAg produces these effects by reacting into receptor sites (possibly Fc receptor sites) on NK cell membrane. The overall significance of our results in relation to hepatitis and hepatocellular carcinoma is discussed.

Adult↗

Changes in superoxide anion production in neutrophils from multitransfused beta-thalassemia patients: correlation with ferritin levels and liver damage.

Superoxide anion production in resting and PMA- or zymosan-stimulated neutrophils was evaluated in 21 beta-thalassemia patients. The results were correlated with ferritin values, hepatitis B virus serum markers, liver pathology, immunoglobulin levels and T-cell subsets. Superoxide anion generation from resting or PMA-stimulated neutrophils was significantly higher in patients than in controls. On the contrary, zymosan-stimulated neutrophils showed reduced superoxide anion production. Increased superoxide anion production in resting neutrophils showed a significant correlation to the values of ferritin. In addition, patients with biopsy-proven chronic liver disease showed significantly increased ferritin levels and superoxide anion production as compared to the remaining patients. No correlation was observed between superoxide anion production and the presence or the absence of hepatitis B virus serum markers, immunoglobulin levels and T-cell subsets. A possible role of interreactions between iron and oxygen radicals in determining liver damage in beta-thalassemia patients is suggested.

Adolescent↗

Raised IgE levels in beta-thalassaemia: correlation with splenectomy and hepatitis B virus infection.

IgE values obtained in 117 beta-thalassaemia patients were significantly higher than in age matched normal subjects. In 31 patients (26.5%) IgE levels were above 2 s.d. of normal values for age, but the frequency of IgE with reaginic activity was lower in patients (5.1%) than in controls (11.9%). The highest values were observed in splenectomized patients who were also positive for one or more serological markers of hepatitis B virus infection. The increase of IgE levels was directly correlated with the number of years after splenectomy, and patients with biopsy proven chronic liver disease had higher IgE levels than those without evidence of liver damage. On the other hand, IgE levels were not correlated with the number of transfusions, age, IgG, IgA, IgM levels or T cell subsets and mitogen responsiveness. These results show that beta-thalassaemia patients develop elevated IgE levels to which splenectomy and hepatitis B virus infection contribute in a synergistic manner.

Adolescent↗

T lymphocytes in children with recurrent respiratory infections: effect of the use of thymostimulin on the alterations of T-cell subsets.

OKT3-, OKT4- and OKT8-positive cells were estimated in 303 children with recurrent respiratory infections. The patients (selected by a score method) had experienced 13 or more infections a year. Modifications in T-cell subsets were observed in 154 patients (50.8%). Decreased OKT3- and OKT4-positive cells were present in 80 children (26.4%), while 74 patients (24.4%) showed normal values of OKT3-positive cells but decreased OKT4- and increased OKT8-positive cells. An attempt at treatment with thymostimulin was undertaken in a group of randomly chosen children with modifications in T-cell subsets. The use of thymostimulin induced the treated children more readily than the untreated ones to show improvement in both the score for respiratory infections and the distribution of T-cell subsets.

Antibodies, Monoclonal↗

A mechanism for liver cell injury in viral hepatitis: effects of hepatitis B virus on neutrophil function in vitro and in children with chronic active hepatitis.

Neutrophil function was studied by nitroblue tetrazolium (NBT) reduction, superoxide anion (O2-) production, chemotaxis, and bactericidal activity in 9 children with HBsAg-positive chronic active hepatitis (CAH). NBT reduction and O2- production were higher in resting neutrophils from the children with CAH than from the controls, but the production of O2- was depressed after stimulation with zymosan. No defect was observed in both random and direct locomotion, whereas a significant decrease was present in bactericidal activity. To evaluate the role of virus components, purified preparations of HBsAg and HBcAg were added to neutrophils from normal children. The incubation with such products induced in vitro the same modifications that were observed in the neutrophils from children with CAH. In addition, defects in phagocytosis and killing of Candida albicans as well as in chemotaxis were demonstrated. The production of O2- was reduced in the neutrophils, stimulated by zymosan and previously opsonized with HBsAg-positive serum.

Adolescent↗

Imbalance in T-cell subsets of five children with congenital agammaglobulinemia and its effect on "in vitro" immunoglobulin synthesis.

OKT3, OKT4 and OKT8 monoclonal antibodies have been used to assess the number of total T cells and of some T-cell subpopulations in five children with Congenital Agammaglobulinemia. A significant increase in OKT8+ cells and parallel decrease in OKT4+ cells were observed in all patients, both as percentages and absolute numbers. by contrast there was no significant difference in the number of total T cells (OKT3+) and OKT3+ OKT4- OKT8- cells between patients and controls. Determinations were made of the synthesis of IgG and IgM by normal B cells cultured with T lymphocytes from patients stimulated by pokeweed mitogen. In all cases a significant decrease in Ig synthesis was observed. These results may be consistent with imbalance in the T regulatory subsets. However, a similar imbalance was observed in a healthy father of one patient, who had no other immunological defect. A normal distribution of T-cell subsets was observed in all the other relatives examined.

Agammaglobulinemia↗

Neutrophyl function in children with chondrodystrophia calcificans congenita: evidence for defective chemotaxis.

Some neutrophil functions (superoxide anion production, bactericidal activity and chemotaxis) have been examined in four children with Chondrodystrophia Calcificans Congenita. Superoxide anion production and bactericidal activity were in the normal range, but a defect in cellular chemotaxis has been proved. This finding account for predisposition to infection which is frequently present in these patients, and may include this disease among those syndromes in which a deficiency of the immune system is associated with growth failure.

Blood Bactericidal Activity↗

Lipid translocation across the human erythrocyte membrane. Regulatory factors.

A simple method based on the differential extraction of lysophosphatidylcholine (LPC) by saline and albumin solutions has been developed to study the factors that influence lipid translocation across intact human erythrocyte membrane. With this assay, the rate of LPC translocation across the bilayer at 37 degrees C was found to be 1.87% h (0.0187 h-1). Identical translocation rates were derived for normal cells and cells in which the ATP was totally depleted, implying that the metabolic state of the cell had no influence. In contrast, the translocation rate was strongly influenced by temperature. Above 21 degrees C, the rate doubled for every 51 degrees C increase in temperature, suggesting an important role for diffusion through the lipid phase. Denaturation of a single major skeletal protein, spectrin, by heating cells to 5 degrees C did not alter the translocation rate. However, oxidative cross-linking of a complex of membrane proteins by treatment with diamide significantly increased the rate of translocation at 37 degrees C. Cholesterol enrichment of the cells decreased the apparent rate of translocation but not the total quantity of LPC translocated. Taken together, these data suggest that lipid translocation across the intact human erythrocyte membrane is not energy dependent, and that it is influenced by the organizational state of both the lipid and protein moieties of the membrane.

Biological Transport↗

Ten-year follow-up study of neonatal hepatitis B immunization: are booster injections indicated?

One hundred and fourteen children, born to HBsAg-positive mothers received in the first year of life passive active prophylaxis for hepatitis B virus (HBV). They have been followed up to 10 years. A booster dose given in a cohort at the 5th year does not seem to increase protection against HBV. No difference in immunological memory was present at the 10th year between the group of children who had received the booster dose at 5 years of age and the group who did not.

Child↗