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Biomedical subjects

R van Furth

Publications and source records attributed to R van Furth.

At least 199 records · Page 11Linked to original sources
PubMed1987

Salmonella-typhimurium-specific difference in rate of intracellular killing by resident peritoneal macrophages from salmonella-resistant CBA and salmonella-susceptible C57BL/10 mice.

The aim of the present study was to determine whether the difference between the rate of intracellular killing of Salmonella typhimurium by macrophages of salmonella-resistant CBA and salmonella-susceptible C57BL/10 mice also holds for other salmonellae and other bacteria species. After in vivo phagocytosis, the initial rate of in vitro intracellular killing of S. typhimurium phagetype 505, S. typhimurium phagetype 510, and S. typhimurium M206 by macrophages of CBA mice amounted always to approximately 1.7 times the value found for macrophages of C57BL/10 mice (p less than 0.001), indicating that the difference in killing efficiency between CBA and C57BL/10 macrophages holds for various strains of S. typhimurium. However, some other salmonella species, i.e., S. dublin and S. heidelberg, as well as E. coli 054 and 02K1+, Listeria monocytogenes EGD and L347, and Staphylococcus aureus were killed equally efficiently by macrophages of both mouse strains. These findings indicate that the difference between the rates of intracellular killing by macrophages of salmonella-resistant CBA and salmonella-susceptible C57BL/10 does not hold for several other bacteria species and thus might be specific for S. typhimurium. Subsequent experiments showed that the in vivo proliferation of S. typhimurium 510 in the first 2 days after i.v. injection was 2.0-fold to 3.0-fold higher in the spleens and livers of C57BL/10 mice than in those of CBA mice, whereas the in vivo proliferation of S. dublin and S. heidelberg was between 1.0-fold to 1.4-fold higher in the C57BL/10 mice. These findings suggest that the differences between the rate of in vitro intracellular killing of salmonella by CBA and C57BL/10 macrophages are reflected in differences in the rate of in vivo proliferation of these microorganisms in CBA and C57BL/10 mice. To gain insight into the involvement of the oxidative metabolism of CBA and C57BL/10 macrophages in the difference in the rate of intracellular killing of S. typhimurium, the O2 consumption and H2O2 release by resident peritoneal macrophages was determined. The amplitudes of the respiratory burst and the release of H2O2 was identical in macrophages of the two mouse strains after triggering by either preopsonized heat-killed S. typhimurium or phorbol myristic acetate. These findings indicate that the mouse species-associated difference in the intracellular killing of S. typhimurium is not caused by a difference in the oxidative metabolism of CBA and C57BL/10 macrophages.

Animals↗
PubMed1987

Quantitative immunocytochemical characterization of mononuclear phagocytes. I. Monoblasts, promonocytes, monocytes, and peritoneal and alveolar macrophages.

The purpose of the present study was to compare the phenotype of tissue macrophages with that of their precursors in the bone marrow and blood. The phenotype was determined on the basis of the quantitative binding of monoclonal antibodies to cell-surface antigens (antigen F4/80, complement receptor III, Fc receptor II, Ia antigen, common leukocyte antigen, and Mac-2 and Mac-3 antigens) on individual mononuclear phagocytes. Monoclonal antibody binding to cells, detected by the biotin-avidin immunoperoxidase procedure, was quantitated by cytophotometric determination of the amount of enzyme reaction product on cells. The results of this quantitation are expressed as the median of the specific absorbance per unit of cell-surface area (0.25 micron2) and per cell. Shortly after collection of the mononuclear phagocytes, binding of all monoclonal antibodies except those directed against the common leukocyte and Mac-2 antigens to peritoneal macrophages was enhanced compared with binding to blood monocytes; for alveolar macrophages we found reduced binding of monoclonal antibodies F4/80 and M1/70 (complement receptor III) and enhanced binding of monoclonal antibodies with specificity for the common leukocyte antigen and Mac-2 and Mac-3 antigens. The results obtained with cultured mononuclear phagocytes show that during the development from monoblast to tissue macrophages, monoclonal antibody binding to the various types of mononuclear phagocyte, expressed per unit of cell-surface area, was not significantly altered except that of M3/38 (Mac-2 antigen) to peritoneal macrophages and that of F4/80 and M1/70 (complement receptor III) to alveolar macrophages. Expressed on a per cell basis, the results show an increase in the binding of all monoclonal antibodies except those directed against the Fc receptor II and Mac-3 antigen during the development from promonocytes to peritoneal macrophages; binding of most monoclonal antibodies to alveolar macrophages was considerably lower than that to blood monocytes. It is concluded that the expression of the various cell-surface antigens alters during mononuclear phagocyte differentiation. The expression changed also during culture, although distinct patterns of alteration could not be distinguished.

Animals↗
PubMed1987

Regulation of granulocyte responses in the blood and peritoneal cavity of CBA and B10 mice during an acute inflammation.

The regulatory mechanisms that determine the course of an inflammation induced by an intraperitoneal injection of kaolin were investigated in Listeria-susceptible CBA and Listeria-resistant B10 mice. The magnitude of the granulocyte inflammatory response in the peritoneal cavity was high in B10 mice (area under the curve; AUC0-48 h: 210.9 x 10(6) granulocytes/mouse x h) and lower in CBA mice AUC0-48 h: 136.8 x 10(6) granulocytes/mouse x h), whereas the reverse was seen for the granulocyte response in the peripheral blood (AUC0-48 h: 30.5 and 80.7 x 10(6) granulocytes/mouse x h, respectively). With respect to the presence of humoral factors that affect the number of granulocytes in the circulation, sera of both mouse strains sampled 24 h after the kaolin injection had granulocytosis-inducing effect in CBA recipient mice and did not induce a response in the B10 recipient mice. This divergent sensitivity to serum factors inducing granulocytosis is consistent with the difference in the blood granulocyte response of B10 and CBA mice but does not explain the divergent inflammatory responses in the peritoneal cavity. Computer simulation showed that at least two factors must be taken into consideration to explain the differences in the inflammatory response, i.e., a factor regulating the release of granulocytes from the bone marrow and a factor governing the rate of granulocyte efflux from the site of inflammation.

Animals↗
PubMed1987

Cell surface characteristics and DNA content of macrophages in murine bone marrow cultures. A study using simultaneous scanning electron microscopy and fluorescence microscopy.

An instrument combining scanning electron microscopy (SEM) and light microscopy (LM) was used to study the cell surface characteristics and DNA content of macrophages in murine bone marrow cultures. After a quantitative Feulgen DNA staining, the DNA content of the individual macrophages was measured and their cell surface morphology was studied immediately thereafter with the SEM part of the instrument. The cells were divided into six groups according to the number of microvilli and/or microridges present on their surface. A proportion of macrophages showed a DNA content more than occurs in diploid cells, which could indicate a future division. No special surface morphology could be detected in this cell type.

Animals↗
PubMed1987

The course of Legionella pneumonia in guinea pigs after inhalation of various quantities of L. pneumophila.

The course of legionella pneumonia in guinea pigs after infection with various quantities of virulent L. pneumophila serogroup 1 organisms by aerosol exposure was investigated. The clinical course, histopathological characteristics, manifestations in the lungs and clearance of the legionella organisms from the lungs and spleen were followed. Four groups were exposed to 4.3 X 10(4), 4.7 X 10(5), 5.0 X 10(6) and 1.0 X 10(8) aerosolized legionellae, respectively. The most striking clinical symptoms were fever and weight loss, which were found in 67-94% and 33-100% of the animals, depending on the dose of L. pneumophila organisms administered. Spontaneous death occurred only in animals receiving the highest dose and always within 10 days. All animals exhibited exponential growth of legionella organisms in the lungs. Maximal growth occurred 5 to 7 days after exposure and varied from 9.3 X 10(7) to 7.4 X 10(8) organisms/both lungs. Twenty-two days after exposure, legionellae could still be cultured from lung tissue. Between 2 and 7 days after exposure, the spleen cultures were positive for legionellae in 41% of the animals receiving the lowest dose and in 83% of all other animals; legionellae could no longer be cultured from spleen tissue after day 7. Depending on the dose, peripherally localized areas of bronchopneumonia increased in size with time, tending to become confluent lobar pneumonia. The microscopic changes were not related to the number of inhaled organisms. In the cellular infiltrate, PMN predominated until day 7 and macrophages thereafter. Seroconversion was found in all animals that survived greater than 7 days. The present animal model closely mimics the course of events in human legionella pneumonia, thus enabling us to further study the factors involved in host resistance against legionella as well as the efficacy of various antimicrobial agents in normal and immunosuppressed animals.

Animals↗
PubMed1987

Effects of tissue-type plasminogen activator on Staphylococcus epidermidis--infected plasma clots as a model of infected endocardial vegetations.

The ability of tissue-type plasminogen activator (t-PA) to enhance the effect of antibiotics in the treatment of bacterial endocarditis was studied using plasma clots infected with Staphylococcus epidermidis as a model of infected endocardial vegetations. A concentration-dependent lysis of the infected plasma clots was induced by t-PA, as shown by the decrease in the weight of the clots, a decrease in the amount of incorporated 125I fibrin, as well as the release of staphylococci from the clots into the incubation fluid. The addition of cloxacillin to the incubation medium in various concentrations led to a concentration-dependent decrease of the number of S. epidermidis in the clots. The presence of t-PA did not enhance the antibacterial effect of cloxacillin. It is concluded that the lysis induced by t-PA might enhance the effect of treatment of endocarditis by reducing the size of endocardial vegetations but not by enhancement of the effect of antibiotics on the bacteria embedded in the vegetations.

Animals↗
PubMed1987

Opsonic and physicochemical characteristics of intravenous immunoglobulin preparations.

The composition and opsonizing activity of five commercially available immunoglobulin preparations for intravenous use (Venoglobulin I, Venilon, Gammagard, Polyglobin, and Sandoglobulin) were studied. The composition of these preparations does not differ very much as far as total protein, immunoglobulin class and IgG subclass concentrations are concerned. The only exceptions were that Veniglobulin I, Gammagard and Sandoglobulin contain IgA, which might cause side effects in patients with anti-IgA antibodies, Gammagard contains very little IgG4, and Venilon and Polyglobin contain no and almost no IgG3, respectively, which might explain their very low opsonic activity. It was found that Venilon and Gammagard activate complement in the ready-for-infusion state. The opsonic activity of Venoglobulin I, Sandoglobulin and Gammagard is about equal to that of inactivated serum: Staphylococcus aureus, Escherichia coli with K antigen, Streptococcus pyogenes and Streptococcus group B are well opsonized and E. coli without K antigen and Streptococcus pneumoniae are poorly opsonized.

Blood Bactericidal Activity↗
PubMed1987

Quantitative effect of granulocytes on antibiotic treatment of experimental staphylococcal infection.

The quantitative relation between granulocytopenia and antibiotic treatment was established for a short-term Staphylococcus aureus infection in the thighs of mice, using rifampin, benzylpenicillin, and erythromycin. Granulocytopenia was induced by total-body irradiation; the number of granulocytes decreased gradually during the first 5 days after irradiation to 10% of the number in normal mice. Experimental infections were established on each of the 5 days after irradiation. In animals not treated with antibiotics, the number of granulocytes in peripheral blood and the number of CFU at the site of infection exhibited a strong negative correlation. The influence of granulocytes on the effect of antibiotics on the number of CFU differed for the three antibiotics. For erythromycin the slope of the dose-effect relation was rather flat, but a decrease in the number of granulocytes caused a significant, nearly parallel, shift in the dose-effect relation, resulting in an increase in the number of CFU. For benzylpenicillin the slope of the dose-effect relation for normal mice was also flat, but as the number of granulocytes decreased the slope became significantly steeper, resulting in a diminishing influence of granulocytes at higher dosages. For rifampin the slope of the dose-effect relation, which was already steep for nonirradiated animals, increased significantly. Here too the effect of granulocytes decreased as the dose increased.

Agranulocytosis↗
PubMed1987

Microphotometric quantitation of the reaction product of several indirect immunoperoxidase methods demonstrating monoclonal antibody binding to antigens immobilized on nitrocellulose.

The nitrocellulose model and microphotometry were used to investigate whether in immunoperoxidase cytochemical methods the amount of final reaction product reflects the amount of cell surface antigen. The results obtained with four cytochemical peroxidase methods, i.e., those using diaminobenzidine/H2O2 (DAB/H2O)2, DAB/H2O2/COCl2, DAB/H2O2/imidazole, and silver intensification of the DAB end product, were compared first. The quantitative DAB/H2O2/imidazole method proved to be the most sensitive and was selected for further studies. Cell surface antigens prepared by solubilization of peritoneal macrophages with octyl-beta-D-glucopyranoside were immobilized on nitrocellulose. Monoclonal antibody binding to these cell antigens was detected by peroxidase immunocytochemistry. Comparison of the sensitivity of the indirect immunoperoxidase and the biotin-(strept)avidin immunoperoxidase methods on the basis of the highest detectable dilution of a cell lysate showed that these methods were equally sensitive. A linear relationship between the absorbance of the peroxidase reaction product and the amount of cell lysate immobilized on nitrocellulose was found for all three indirect immunoperoxidase methods. This proves that the amount of final immunocytochemical peroxidase reaction product is proportional to the amount of antigen in cell lysates. However, the relative expression of antigens in intact cells differs from that in cell lysates. Therefore, the present method to solubilize cells and immobilize cell antigens cannot be used to quantitate the antigen content of cells.

Animals↗
PubMed1987

Quantitative immunocytochemical characterization of mononuclear phagocytes. II. Monocytes and tissue macrophages.

The purpose of the present study was to compare the monoclonal antibody (Mab) binding patterns of various tissue macrophages with each other and with blood monocytes. To allow recovery from the effects of the isolation procedure, or to obtain purified populations, macrophages were cultured for 24 hr and 48 hr. For comparison, blood monocytes were also cultured for 24 hr and 48 hr. Mab binding to individual cells, detected by the biotin avidin immunoperoxidase method, was quantified cytophotometrically and the results expressed as the median of the specific mean absorbance per 0.25 micron2 cell surface area or as specific integrated absorbance per cell. Analysis of the quantitative data in relation to the results of subjective evaluation of the peroxidase reaction product, demonstrating Mab binding to cells, yielded three classes for description of the intensity of antigen expression by cells: weak (specific mean absorbance per unit cell surface less than 0.07), moderate (values between 0.07 and 0.14), and intense (values more than 0.14). No matter how the results were expressed, comparison of the Mab binding patterns of macrophages with those of blood monocytes showed that spleen macrophages bound significantly less F4/80 and more M5/114 (Ia antigen). Kupffer cells and skin macrophages bound either approximately the same amount or considerably less of the various Mabs than monocytes did. Pulmonary tissue and alveolar macrophages bound significantly more 30.G.12 (leucocyte antigen), M3/38 (Mac-2 antigen), and M3/84 (Mac-3 antigen) and comparable amounts or considerably less of the other Mabs than the monocytes did. Peritoneal macrophages bound significantly more F4/80, M1/70 (complement receptor III), and 2.4.G.2. (Fc receptor II) and comparable amounts or considerably less of the other Mabs than monocytes did. It is concluded that macrophages from different organs and different anatomical sites within one organ differ from one another, for example, peritoneal macrophages do not resemble any other population of macrophages and alveolar macrophages do not resemble pulmonary tissue macrophages, and differentiation of blood monocytes into tissue macrophages does not show a distinct pattern.

Animals↗
PubMed1987

Cell kinetic analysis of a murine macrophage cell line.

For the present study, which was performed to find a reliable method suitable for determination of the cell kinetic parameters of a continuous cell line, use was made of the macrophage cell line J774.1. The doubling time of the cell population was approximately 27 h. The continuous labeling curve showed that all the cells divide and almost no quiescent cells occur. The cell-cycle time as determined from the curve of the labeled cells in mitosis, the course of the stathmokinetic index, and time-lapse videorecordings, was about 19 h. The discrepancy between the population doubling time and the cell-cycle time must be due to death and disintegration of cells during culture in vitro. The results indicate that the doubling time of a cell population is not a reliable parameter to determine the kinetics of a population of continuously proliferating cells and that determination of the course of the stathmokinetic index offers a rapid and simple method to establish the cell-cycle time reliably.

Animals↗
PubMed1986

Distribution of blood monocytes between a marginating and a circulating pool.

Studies on the blood volume of SPF Swiss mice in the steady state led to reevaluation of the distribution of monocytes in the blood compartment. It was concluded from the results of our calculations that monocytes entering the circulation are distributed over a circulating and a marginating pool. The circulating pool accounts for approximately 40%, and the marginating pool or approximately 60% of the population of peripheral blood monocytes.

Animals↗
PubMed1986

Differences in initial rate of intracellular killing of Salmonella typhimurium by granulocytes of Salmonella-susceptible C57BL/10 mice and Salmonella-resistant CBA mice.

The contribution of granulocytes to differences in the innate susceptibility of mouse strains to infection by Salmonella typhimurium was assessed on the basis of the size and composition of the inflammatory exudate after i.p. injection of bacteria and the intracellular killing of the bacteria by exudate peritoneal cells and blood granulocytes of resistant CBA and susceptible C57BL/10 mice. The increase in the numbers of both peritoneal granulocytes and macrophages 24 hr after i.p. injection of various numbers of live S. typhimurium was two to four times higher in C57BL/10 mice (p less than 0.05) than in CBA mice. However, despite the larger number of phagocytes in the inflammatory exudate, the numbers of viable S. typhimurium in the peritoneal cavity 24 hr after injection was higher (p less than 0.01) in C57BL/10 mice than in CBA mice. Because the proportion of noningested bacteria was similar in the two mouse strains (less than 30%), these findings indicate a difference in the rate of intracellular killing of the bacteria by exudate peritoneal cells (greater than 75% granulocytes) of the two mouse strains. Subsequent determination of the initial rate of intracellular killing (Kk) of S. typhimurium revealed that after phagocytosis of the bacteria in vivo, exudate peritoneal granulocytes (harvested 24 hr after i.p. injection of 10(3) live S. typhimurium) of CBA mice killed S. typhimurium twice as efficiently (Kk = 0.014 min-1; p less than 0.01) as exudate granulocytes of C57BL/10 mice (Kk = 0.008 min-1) did. Similarly, the initial rate of intracellular killing of the ingested S. typhimurium by blood granulocytes of CBA mice (Kk = 0.017 min-1) was two times higher (p less than 0.01) than that of C57BL/10 mice (Kk = 0.007 min-1). These findings may be specific for S. typhimurium, because L. monocytogenes were killed with equal efficiency by exudate granulocytes and blood granulocytes of these mouse strains (p greater than 0.20). The results of the present study are relevant with respect to the innate resistance of mice to S. typhimurium, particularly during the initial phase of infection when the inflammatory exudate contains predominantly granulocytes.

Animals↗
PubMed1986

Quantitative study of enzyme immunocytochemical reactions performed with enzyme conjugates immobilized on nitrocellulose.

The nitrocellulose binding assay was used for quantitative studies on the cytochemical reactions for the three enzymes most frequently used in immunocytochemistry. The results show a linear relationship between the amount of enzyme immobilized on nitrocellulose and the amount of the enzyme reaction product. The similar course of the formation of the reaction product after DAB/H2O2 staining for peroxidase immobilized on nitrocellulose and for immunoperoxidase labeled cells indicates a linear relationship between the amount of enzyme-coupled antibodies bound to cells and the amount of enzyme reaction product. Furthermore, a mild acid treatment for the abolition of endogeneous peroxidase activity in tissues and cells applicable to immunoperoxidase staining procedures is proposed.

3,3'-Diaminobenzidine↗
PubMed1986

Opsonic activity and composition of five intramuscular gammaglobulin preparations.

The opsonic activity of five immunoglobulin preparations for intramuscular and subcutaneous substitution therapy was investigated. Various strains of Staphylococcus aureus, Escherichia coli, Streptococcus pyogenes, group B streptococci, Streptococcus pneumoniae and Haemophilus influenzae were used. In addition, the concentration of unsplit IgG molecules, IgG fragments and IgG aggregates in these preparations was determined. With the exception of H. influenzae, all preparations opsonised the bacteria studied almost as well as inactivated serum. All preparations contained an appreciable amount of aggregates, but IgG fragments were not found.

Escherichia coli↗
PubMed1986

The effect of glucocorticosteroids on bone marrow mononuclear phagocytes in culture.

The effect of hydrocortisone and dexamethasone both in vivo and in vitro was studied in mouse bone marrow cultures in methylcellulose and in two liquid culture systems, one using Leighton tubes with a flying coverslip to grow adherent colonies and the other using Teflon culture bags to obtain suspension cultures. Although the total number of nucleated bone marrow cells was not greatly influenced by glucocorticosteroid treatment of the mice, a marked decrease in the number of colony-forming units and of mononuclear phagocytes was observed. Inhibition of colony growth in methylcellulose and of growth of mononuclear phagocytes in Teflon culture bags also occurred when glucocorticosteroids were added to in vitro cultures. Both drugs caused an almost complete inhibition of the growth of adherent colonies, and 3H-thymidine labeling of the cells was correspondingly low. When the glucocorticosteroids were added to cultures pre-incubated for 5 days in the presence of conditioned medium, the 3H-thymidine labeling of macrophages and promonocytes was markedly reduced, whereas there was no change in the labeling of monoblasts.

Animals↗
PubMed1986

Effect of penicillin G on Staphylococcus aureus phagocytosed by human monocytes.

The effect of penicillin G on Staphylococcus aureus after phagocytosis by human monocytes was compared with the effect on nonphagocytosed bacteria. Intracellular S. aureus was obtained by incubation of monocytes and preopsonized S. aureus for 3 min at 37 C; phagocytosis was stopped by cooling the test tubes in crushed ice. Nonphagocytosed bacteria were removed by differential centrifugation followed by two washes of the cells. Morphological studies with lysostaphin confirmed the intracellular localization of approximately 70% of the bacteria after this procedure. The antibacterial activity of penicillin G was assessed by incubation of monocytes containing ingested S. aureus with the drug at 37 C, and determination of the number of viable cell-associated bacteria was assessed microbiologically at various intervals. Inactivation of the intracellular killing mechanism of the monocytes by omission of serum during incubation allowed measurement of the effect of penicillin G apart from the bactericidal activity of the cells. The effect of penicillin G on nonphagocytosed bacteria was assessed after incubation of preopsonized S. aureus in medium with the drug. The results show that the effect of penicillin G on intracellular S. aureus is two to seven times stronger than the effect on nonphagocytosed S. aureus.

Humans↗