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Meir Djaldetti

Publications and source records attributed to Meir Djaldetti.

13 recordsLinked to original sources

Functional hypersplenism in mice induced by adoptive transfer of syngeneic spleen cells.

Investigation of peripheral blood cell count alterations in cases with hypersplenism, and an understanding of the relationship between splenic function and hematopoietic cell production require suitable experimental animal models. Previously described methods are either traumatic or require surgical intervention. We suggest a relatively simple method for achievement of a state mimicking hypersplenism in mice by intraperitoneal inoculation of syngeneic spleen cells. Mice were inoculated intraperitoneally with 3 x 10(7) splenocytes suspended in 0.3 ml phosphate buffered saline (PBS). After 2 months, the inoculated animals showed a progressive decrease in the peripheral white blood cell (WBC) counts and hyperplastic bone marrow that persisted until the experimental end point (7 months). Five days after inoculation of splenocytes stained with carboxy-fluorescein diacetate succinimidyl ester (CFSE), the majority of the stained cells was present in the peritoneal cavity (33%) and in the liver (13%), whereas the percentage of stained cells in the peripheral blood and the spleen cell suspension was negligible. The mitogen response of the peripheral blood mononuclear cells (PBMC) from treated mice to concanavalin A (Con A) remained unaltered. Splenocyte-inoculated mice that were further splenectomized did not show leukocytosis after splenectomy, as was observed in animals in which the spleen was removed without any pretreatment. The lack of any signs of discomfort in animals from the study group, in comparison with the visibly ill appearance and even death of mice in which hypersplenism was achieved by repeated injections of methylcellulose (MC), which served as controls, favors the convenience of the method.

Adoptive Transfer↗

Effect of pravastatin, simvastatin and atorvastatin on the phagocytic activity of mouse peritoneal macrophages.

Since cholesterol and lipid content may affect cell membrane fluidity, we assumed that treatment of mice with lipid lowering statins would enhance the engulfing capacity of their macrophages. Four groups of animals were examined. Group I-treated with pravastatin, group II--with simvastatin--both drugs in a dosage of 40 mg/kg daily, 5 days/week for a total of 3 weeks. Mice in group III received atorvastatin 5 mg/kg for the same time period. Group IV--untreated animals serving as controls. The phagocytic capacity of the peritoneal macrophages was evaluated by their ability to engulf latex particles. In addition, the mitogen response of the peripheral blood mononuclear cells (PBMC) and splenocytes to Con A and PHA was examined. Compared to the controls, the percentage of phagocyting cells in pravastatin-treated mice was enhanced by 18%, with simvastatin--by 24% and in atorvastatin-treated animals by 8%. The three statins increased the phagocytic index by 79.5%, 88.8% and 62%, respectively. The mitogen response of splenocytes from mice treated with the three statins to Con A increased by 68%, 48% and by 40%, respectively. Compared with the controls the response to PHA was higher in animals treated with pravastatin (84%), simvastatin (73%) and atorvastatin (57%). The response of PBMC from statin-treated animals to both mitogens did not differ from that of the controls. The results suggest that statins, at least those hereby investigated, may exert a beneficial effect on the immune function of the macrophages.

Animals↗

Phagocytic capacity and apoptosis of peripheral blood cells from patients with iron deficiency anemia.

Following clinical observations that patients with iron deficiency anemia (IDA) are more susceptible to infections than non-anemic individuals, the phagocytic capacity and number of apoptotic peripheral white blood cells (PWBC) from patients with IDA were examined. PWBC from 15 patients with IDA and from 18 healthy donors were incubated with various doses of iron. Phagocytosis was examined using latex particles and apoptosis was evaluated by a flow cytometric assay using propidium iodide staining. The percentage of phagocyting polymorphonuclear cells was lower in IDA patients compared to that of the controls. However, there was no difference in the percentage of phagocyting monocytes from individuals of both groups. The number of latex beads engulfed by each polymorphonuclear or monocyte was lower in IDA patients. Incubation with 100 microg% of iron did not affect the phagocytic ability of both cell types in IDA patients, but increased that of control cells. Incubation with 300 microg% of iron caused an increase in the phagocytic capacity of patients' cells and a decrease in that function in cells from controls. Higher dose (500 microg%) induced suppression of phagocytosis in cells from both groups. There was no difference in the number of apoptotic cells from individuals of both groups. Apoptosis of polymorphonuclears, but not mononuclear cells from both controls and IDA patients showed a linear dependency on the iron concentration in the medium. It is possible that the impaired phagocytic capacity of the PBWC found in patients with IDA contribute to the increased susceptibility to infections observed in these individuals.

Anemia, Iron-Deficiency↗

In vitro effect of statins on cytokine production and mitogen response of human peripheral blood mononuclear cells.

The in vitro effect of the hydrophilic statin - pravastatin - and three hydrophobic statins - atorvastatin, lovastatin and simvastatin - on the production of IL-1beta, IL-1ra, IL-2, IL-6 and IFN-gamma by human peripheral blood mononuclear cells (PBMC) and their response to mitogens was examined. Lovastatin and simvastatin increased the production of IL-1beta in a dose dependent manner and reduced secretion of IL-1ra at high concentration. These two statins exerted a dose dependent inhibitory effect on IL-2 production and reduced the secretion of IFNgamma at high dose. Atorvastatin did not affect IL-1beta, but suppressed IL-1ra, IL-2 and IFNgamma production. Atorvastatin, lovastatin and simvastatin caused a dose dependent inhibition of mitogen-induced proliferation of PBMC. IL-6 production was not affected by any one of the statins. While pravastatin caused a slight reduction in the proliferative response of PBMC to PHA, it did not affect either their response to Con A and PWM, or the secretion of cytokines tested.

Atorvastatin↗

Ultrastructural features of bone marrow cells from patients with acquired sideroblastic anemia.

The ultrastructural findings of the bone marrow cells from 15 patients with acquired sideroblastic anemia are presented. The red cell precursors from all patients showed the presence of electron-dense material in the mitochondria, representing most probably iron deposits. A great number of these mitochondria were completely destroyed. The erythropoietic precursors from one of the patients showed markedly elongated mitochondria that measured up to 3 microm. In addition numerous cytoplasmic vacuoles were observed. The red cell precursors from 60% of the patients showed signs of dyserythropoiesis, such as incomplete nuclear division and nuclear distortion. The polymorphonuclears from 47% of the patients presented nuclear abnormalities expressed as nuclear bridges, appendices, and blebs. In addition, phagocytosis of red blood cells was observed. The results of the study underline the advantages of the transmission electron microscope examination in visualization of intricate alterations in hematopoietic cells that cannot be detected with a light microscope.

Aged↗

In vitro cytokine production in patients with iron deficiency anemia.

The in vitro production of interleukin (IL)-1beta, IL-2, IL-6, IL-10, and tumor necrosis factor alpha (TNFalpha) by peripheral blood mononuclear cells (PBMC) from 20 patients with iron deficiency anemia (IDA) was examined before and after iron supplementation and compared to values obtained for PBMC from healthy controls. A significant decrease in IL-2 production was observed in IDA patients, whereas the secretion of the other cytokines did not differ from that of controls. Addition of iron to the culture medium did not affect the secretion of IL-2 and IL-1beta, but caused an increase in IL-6, IL-10, and TNF-alpha production. Since a deficiency in IL-2 production plays a role in the pathogenesis of certain infectious and malignant diseases, the results of the present study may explain in part the increased susceptibility to infections observed in patients with IDA.

Anemia, Iron-Deficiency↗

In vitro immune response of human peripheral blood cells to vitamins C and E.

Since oxygen free radicals exert a noxious effect on cell functions, the purpose of the study was to examine the influence of the antioxidant vitamins C and E on the phagocytic capacity, apoptotic death, production of TNFalpha and IL-10 by human peripheral blood cells. In addition, an attempt to find a correlation between the effect of these vitamins on apoptosis and DNA synthesis was carried out. Peripheral white blood cells obtained from 27 healthy volunteers were incubated for 24 hr without and with vitamins C and E at doses extrapolated from clinical practice. Incubation of cells with vit. C caused a significant increase in the number of latex particles internalized by each individual polymorphonuclear cell, but not by monocytes. Both vitamins did not change the number of cells capable for phagocytosis. By the method of propidium iodide staining for detection of apoptosis, incubation of the cells with 0.2 mg/mL vit. C for 24 hrs caused a 39% increase in the percentage of apoptotic cells, as compared to those kept at the same incubation conditions without vitamin. 0.125 mg/mL of vit. E did not affect the percentage of apoptotic cells. On the other hand, applying the caspase-3 method for apoptosis detection, vitamins C and E did not affect the caspase-3 activity. Both vitamins caused an inhibition of 3H-TdR incorporation, which was dose-dependent for vit. C. Concentrations of the vitamins lower than those mentioned above did not alter DNA synthesis. While TNFalpha production was not affected by both vitamins, the spontaneous secretion of IL-10 was dose-dependently reduced by vit. C but remained unaltered following incubation with vit. E. The results, although observed in vitro, might be of importance when those vitamins are administered to healthy subjects.

Antioxidants↗

The relationship between partial splenectomy and peripheral leukocyte count.

BACKGROUND: Partial splenectomy is accepted as a treatment modality for hypersplenism permitting preservation of the spleen functions. Since a prominent leukocytosis is a marked event after total splenectomy, it was the aim of the present study to compare the peripheral white blood cell counts (PWBC) and immune response in mice following partial and total splenectomy. MATERIALS AND METHODS: Four groups of animals were included in the study: mice in which 70% of the spleen was removed, animals that underwent total splenectomy, mice with sham operation, and a group of mice that served as controls. The proliferative response of peripheral blood mononuclear cells, peritoneal cells, and splenocytes was examined using concavalin (Con) A. RESULTS: In distinction from marked leukocytosis observed in mice after total splenectomy, in partially splenectomized mice the PWBC counts did not show any significant increase during a follow-up period of up to 2 months after surgery. The mitogen response of the mononuclear cells to Con A in partially splenectomized mice was similar to that of controls, while in animals after total splenectomy, it was increased in cells from the peripheral blood and decreased in those from the peritoneum. CONCLUSIONS: The results indicate that removal of as much as about 70% of the spleen in mice is sufficient to maintain a normal PWBC count, suggesting a regulatory role of the spleen remnant on the PWBC production. The normal mitogen response of the cells to Con A indicates that the spleen rudiment preserves at least a part of the immune activity of the intact spleen.

Animals↗

Ultrastructure of mouse striated muscle fibers following pravastatin administration.

To examine the effect of pravastatin administration on striated muscle ultrastructure, 10 BalbC mice were given pravastatin 40 mg/kg/day for 3 weeks. At the end of the study, blood was withdrawn for evaluation of the serum creatine phospho-kinase (CPK) level and the muscles of the hind legs, as well as the heart and liver of the animals were examined with a light and transmission electron microscope. After treatment with pravastatin the results showed a 101% increase in serum CPK level in comparison to untreated controls. Hematoxillin-eosin stained tissues of pravastatin treated mice did not show any abnormal findings. While the ultrastructure of the heart and liver of the treated animals appeared normal, the muscle fibers showed a marked alterations of the mitochondria, which were increased in size compared to those of the controls. The cristae were heavily damaged and even completely destructed, giving the mitochondria appearance of empty vacuoles. The findings are in favor of a specificity of pravastatin for striated muscles.

Animals↗

Phagocytosis--the mighty weapon of the silent warriors.

Professional phagocytes, comprising polymorphonuclear neutrophils and monocyte/macrophage cells, play an important role in the host defense. Any defect in their function exposes the organism to microbial intruders terminating in fatal diseases. The functional responses of the phagocytes to bacterial and fungal infections include chemotaxis, actin assembly, migration, adhesion, aggregation, phagocytosis, degranulation, and reactive oxygen species production. Superoxide generation by phagocytic NADPH oxidase is an imperative step toward bacterial killing. Phagocytes participate in inflammatory reactions and exert tumoricidal activity. They are supported by serum factors such as immunoglobulins, cytokines, complement, the acute phase reactant C-reactive protein, production of antibacterial proteins, and others. In addition to their principal task to eliminate bacteria, they are engaged in removing damaged, senescent, and apoptotic cells. Engulfed cell debris, large particles such as latex beads, fat, and oil droplets, are examples of phagocytic activity illustrated in the present review with transmission and scanning electron microscope micrographs. Numerous factors, such as diseases and stressful conditions, affect the engulfing activity of the professional phagocytes. Our experience regarding the impaired phagocytic capacity of cells in patients with diabetes and chronic renal failure is discussed. The results obtained in our laboratory from experiments detecting the effect of strenuous physical exercise, hypothermia, fasting, and abdominal photon irradiation on the phagocytic capacity of human polymorphonuclear neutrophils and rat peritoneal macrophages are hereby summarized and the reports on those subjects in the recent literature are reviewed. A variety of assays are applied for quantifying phagocytosis. Flow cytometry based on incubation of phagocytic cells with fluorescent conjugated particles and measuring the amount of fluorescence as an indicator of the engulfing capacity of the cells is a useful method. A direct visualization of the ingested particles using light or electron microscopy is a valuable tool for estimation of phagocytic function. In our hands, the use of semithin sections of embedded phagocytes following their incubation with latex particles provided satisfactory results for measuring the total number of phagocytic cells, as well as the internalizing capacity of each individual cell. Microbiological assays, the nitroblue tetrazolium test, quantitation of antibody- and antigen-mediated phagocytosis, as well as methods reviewed in detail in other reports are additional applications for determination of this intricate process.

Animals↗

Factor(s) released from irradiated B-CLL cells induce apoptosis in leukemic lymphocytes.

Photon irradiation of peripheral blood lymphocytes from 25 patients with untreated B-chronic lymphocytic leukemia (B-CLL) induced an increase in apoptotic response by 270%. No significant increase in apoptosis was observed after irradiation of peripheral blood mononuclear cells from 15 healthy volunteers. Supernatants (sups) derived from irradiated leukemic cells incubated with non-irradiated autologous cells induced a 75% enhancement in number of apoptotic cells, as compared with sups from non-irradiated CLL cells. The level of tumor necrosis factor alpha, a cytokine known to prevent apoptosis, was reduced in the sups of irradiated CLL cells in comparison to that of non-irradiated lymphocytes. The interleukin (IL)-10 level, an IL reported to induce apoptosis, was similar in the sups of irradiated and non-irradiated lymphocytes from B-CLL patients. No change in IL-2 levels was observed. The significance of these findings and the role of factor(s) in the sups of irradiated leukemic lymphocytes as inducers of apoptosis are discussed.

Aged↗

Are peripheral blood cells from patients with Alzheimer disease more sensitive to apoptotic stimuli?

One of the reasons for the increased susceptibility to infections in patients with Alzheimer disease may be enhanced apoptotic death of their peripheral leukocytes. If this is the case, the enhanced apoptosis may be due to components in the patients' sera or to an increased sensitivity of the cells to apoptotic stimuli. To examine this possibility, the percentage of apoptotic cells in the peripheral blood mononuclear cells (PBMC) from 12 patients with Alzheimer disease was compared with that of 12 age-matched non-demented persons and 12 middle-aged healthy control subjects. In addition, the effect of sera from subjects in the three groups on the apoptosis, interleukin (IL)-1beta, IL-2, IL-6, IL-10, and tumor necrosis factor (TNF)alpha production by peripheral blood cells from healthy control subjects was examined. It was found that the percentage of apoptotic PBMC from patients with Alzheimer disease was higher than that from the remaining two groups. However, incubation of control cells with sera from patients with Alzheimer disease and non-demented elderly persons did not affect the number of apoptotic cells. Sera from patients with Alzheimer disease and non-demented elderly subjects caused an increase in IL-2 and a decrease in IL-10 production by PBMC from middle-aged control subjects but did not affect IL-1beta, IL-6, and TNFalpha secretion, indicating alterations of the immune system related to aging.

Aged↗

Cytokine levels and phagocytic activity in patients with Alzheimer's disease.

BACKGROUND: Clinical observations indicate that patients with Alzheimer's disease show a greater susceptibility to infections. One possible explanation is that this predisposition is due to alterations in their immune system. OBJECTIVE: To investigate this assumption, pro- and anti-inflammatory cytokines, as well as the phagocytic activity and superoxide anion generation was examined in aged individuals with and without Alzheimer's disease. METHODS: The production of IL-1beta, IL-2, IL-6, TNFalpha and IL-10 by peripheral blood mononuclear cells from 12 patients with Alzheimer's disease was compared with that of 12 age-matched individuals without any signs of dementia and 12 middle-aged healthy volunteers who served as an additional control. The engulfing capacity of the phagocytic cells was detected by counting cells containing latex beads and the number of particles internalized by each individual cell. RESULTS: The secretion of IL-2 was markedly low in the demented patients, compared with both elderly and middle-aged subjects. IL-1beta and TNFalpha production was similar in the individuals of the 3 groups. The production of IL-6 and IL-10 was significantly lower when compared to that of the middle-aged, but did not differ between the elderly patients with and without dementia. The phagocytic function of both polymorphonuclear cells and monocytes was decreased in individuals of the elderly groups with a low number of engulfed latex particles by each individual polymorphonuclear cell. The production of superoxide anions was increased only by monocytes from the elderly groups. CONCLUSIONS: The results suggest that although the impaired immune function in patients with Alzheimer's disease is related to the aging process, the significant low IL-2 production in these patients may play a role in their increased susceptibility to infections.

Aged↗