bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of Cyclic Nucleotide Metabolism During Human Monocyte Differentiation
Authors: O'Dorisio M S , Fertel R , Finkler E , Brooks R , Vassalo L
Published: January 2018

Cyclic nucleotide metabolism was studied during human monocyte differentiation. Intracellular cAMP increased 17-fold during in vitro differentiation. This increase was due to an increase in the specific activity of adenylate cyclase and a concomitant decrease in the specific activity of cyclic AMP phosphodiesterase. Monocyte adenylate cyclase activity was stimulated by guanine nucleotide, prostaglandin E1, isoproterenol, and epinephrine. Macrophage adenylate cyclase demonstrated less responsiveness to guanine nucleotide and was refractory to stimulation by prostaglandin E1, and to β-adrenergic receptor agonists.

In contrast to cAMP, total intracellular cGMP levels remained constant. Guanylate cyclase was found predominately in the cytosol of monocytes. The specific activity of soluble guanylate cyclase decreased during differentiation, while particulate activity increased more than 40-fold. Cyclic GMP phosphodiesterase activity remained stable during monocyte differentiation.

The ratio of cAMP:cGMP increased dramatically from 2:1 in monocytes to 9:1 in macrophages suggesting that cAMP may be an important mediator of differentiation, while cGMP metabolism decreases in the fully differentiated nonproliferating macrophage.
Opsonin-Independent Phagocytosis of Surface-Adherent Bacteria by Human Alveolar Macrophages
Authors: Lee David A , Hoidal John R , Garlich Dawn J , Clawson C Carlyle , Quie Paul G , Peterson Phillip K
Published: January 2018

Opsonin-independent mechanisms of phagocytosis may be important in host defense of certain body sites such as the lung, in this study, one such mechanism, “surface phagocytosis,” was investigated by measuring the uptake of unopsonized [3H]-labeled Staphylococcus aureus and Pseudomonas aeruginosa adherent to a plastic surface by human alveolar macrophages (AM) and peripheral blood polymorphonuclear leukocytes (PMN). Efficient uptake of unopsonized bacteria by both cell types was observed. Electron microscopic studies suggested that the manner in which these cell types encounter adherent bacteria is different. While AM appear to gather in organisms at their periphery as they spread out upon the underlying substrate, PMN seem to sweep bacteria up as they move along the plastic surface. Bacterial killing determined by a fluorochrome microassay was decreased by AM compared to PMN. Although the precise mechanism whereby phagocytes recognize unopsonized bacteria adherent to a surface remains to be determined, this aspect of phagocytic cell function may prove to have clinical relevance.
The Separation of Cytotoxic Human Peripheral Blood Monocytes Into High and Low Phagocytic Subsets by Centrifugal Elutriation
Authors: Chiu K M , McPherson L H , Harris J E , Braun D P
Published: January 2018

Mononuclear cells (4 × 108 in a 5-ml volume) were loaded onto a Beckman JE-6B elutriator rotor spinning at 2,000 ± 10 rpm with a flow rate of 10.0 ml/ min. After lymphocytes were removed at flow rates between 10 and 11 ml/min with 500 ml of buffer, the flow rate was increased by 0.5 ml/min/fraction to collect 100-ml fractions. Highly enriched monocytes as judged by nonspecific esterase staining and morphology (70–95%) were found in each of eight fractions collected with flow rates between 11.5 to 15.0 ml/min. When stimulated with phorbol myristic acetate, these fractions mediated equivalent levels of cytotoxicity against 51Cr-labeled Chang liver cell line. Similarly, each mono- cyte-containing fraction was found to mediate the same level of cytotoxicity against antibody-sensitized 51Cr-labeled Chang liver cells. In contrast, cytotoxicity against the natural killer cell-sensitive K-562 cell line was found in only those fractions that contained a high percentage of lymphocytes. The fractions that were enriched in monocytes were found to differ in their ability to ingest latex. Those monocyte fractions that were collected between 11.0–12.0 ml/min consisted primarily of low numbers of latex-ingesting monocytes (<30%). Those monocyte fractions that were collected between 12.5 and 15.0 ml/min consisted primarily of latex-ingesting monocytes (50–70%). These data show that cytotoxic monocytes can be separated by centrifugal elutriation into at least two subsets that can be distinguished by their phagocytic activity.
The Modification of the Oxidative Metabolism of Cells Derived Both Locally and at Distance From the Site of an Acute Inflammatory Reaction
Authors: Bird John , Pelletier Monique , Tissot Michéle , Giroud Jean-Paul
Published: January 2018

During an acute nonspecific inflammatory reaction initiated in the pleural cavity by a nondiffusible stimulus (calcium pyrophosphate crystals), the oxidative metabolism, as measured by chemiluminescence and superoxide release, of cells harvested from both the inflammatory site and at points distant from it was studied.

The oxidative metabolism of peritoneal macrophages, obtained from rats undergoing an inflammatory reaction (pleurisy), demonstrated a transient decrease in activity compared with the resident population when using both zymosan and phorbol myristate acetate as stimulants. This metabolic unresponsiveness induced by inflammation may be related to the concomitant changes in the levels of prostacyclin in the peritoneal cavity. It should be emphasized that the peritoneal cellular composition or number did not change during these events. On the other hand alveolar macrophages from inflamed animals showed no significant changes in their superoxide production or chemiluminescence compared to controls. The precise reason for these inflammation-induced changes is unknown; however the acute nonspecific inflammatory reaction was able to modulate the oxidative metabolism of cells not only at the site of inflammation, but at points distant from it.
Phagocytosis, Chemiluminescence, and Cell Volume of Alveolar Macrophages From Neonatal and Adult Pigs
Authors: Zeidler R B , Kim H D
Published: January 2018

Broncho-alveolar cells lavaged from the lungs of 1- and 7-day-old piglets and adult pigs were examined to determine their alveolar macrophage content, cell size, and ability to phagocytyze emulsified oil droplets and to chemiluminesce in response to opsonized zymosan particles. Alveolar macrophages were identified by specific cytochemical stains and differential cell counts as being 95%, 97%, and 90% of the cell populations, respectively. The cell volume of macrophages from 1-day-old pigs was 383 ± 40 µm3; while the cell volumes from 7 day and adult pigs were 1,660 ± 265 and 1,411 ± 310 µm3. Alveolar macrophage from 1-day-old piglets possessed little ability to engulf oil droplets; however, macrophages from 7-day-old piglets phagocytized these opsonized droplets at a rate equivalent to alveolar macrophages obtained from adult pigs. The phagocytosis of oil droplets by both 7-day and adult alveolar macrophages manifested similar characteristics. Both rates can be described by saturation kinetics, and while temperatures from 37° to 30°C had little effect, the rate declined rapidly between 30° and 16°C. Alveolar macrophages from 1-day-old piglets displayed a lower magnitude of chemiluminescence than cells from 7- day-old and adult pigs and did not always respond to zymosan exposure. On the other hand, 7-day-old and adult pig alveolar macrophages were quite similar in their ability to chemiluminesce. Increasing extracellular glucose from 1 to 20 mM induced a concomitant increase in chemiluminescence. The findings suggest that functionally competent alveolar macrophages emerge in the pig lung approximately 1 week after birth.
Reticuloendothelial System Activated by Fructose-1,6-Diphosphate in Mice
Authors: Tarsi R , Mulieri L , Venturi F , Simonetti N
Published: January 2018

Fructose-1,6-diphosphate (FDP) exerts a marked activity on reticuloendothelial system (RES) functions, increasing in mice the clearance of colloidal carbon. ATP depletion occurring during phagocytic activity is concentrated by FDP.
Influence of Prostaglandin I2 on Fibronectin-Mediated Phagocytosis In Vivo and In Vitro
Authors: Weinberg David A , Weston Lisa K , Kaplan John E
Published: January 2018

This study evaluated the effect of prostaglandin I2 (PGI2)on fibronectin-me- diated macrophage phagocytosis in vivo and in vitro. Phagocytosis measured in vivo in rats by the vascular clearance rate and hepatic localization of gelatinized sheep erythrocytes was inhibited in a dose-dependent manner after intravenous administration of PGI2. Phagocytosis was assessed in vitro in terms of uptake of fibronectin-dependent gelatinized sheep erythrocytes by monolayers of casein-elicited rat peritoneal macrophages. Concentrations of 1 ng/ml PGI2 or greater resulted in inhibition of particle internalization but not attachment to macrophages. This inhibitory effect was enhanced by amino- phylline, a phosphodiesterase inhibitor. PGI2 increased cAMP levels and these were further increased in the presence of aminophylline. These data indicate that PGI2 inhibits macrophage uptake of gelatinized particles and support the idea that this is mediated by increased intracellular levels of cyclic AMP. PGI2 should thus be considered a potential etiologic factor in the phagocytic depression observed in association with thrombosis.
The Response of Human Peripheral Blood Mononuclear Phagocytes to Rheumatoid Arthritis
Authors: Buchan Glenn S , Palmer David G , Glbbins Bruce L
Published: January 2018

The maturity of peripheral blood mononuclear phagocytes (B-MPs) from patients with rheumatoid arthritis (RA), osteoarthritis (OA), and reference (“normal”) subjects was compared. Mononuclear cell isolates from peripheral blood were separated on discontinuous gradients of Percoll into low density (more mature) and high density (less mature) subpopulations. Contrary to expectations, the proportion of immature B-MPs in RA patients was found to be significantly lower than that in reference subjects. In RA patients with synovial effusions the proportion of immature B-MPs approached but did not exceed those found in reference subjects, despite the fact that 31% of these patients displayed a peripheral blood monocytosis. It was concluded that the bone marrow precursor population had adapted to the long-term demand for B-MPs in the course of this chronic inflammatory disease.
Modulation of Mononuclear Phagocyte Cytotoxicity by Alpha-Tocopherol (Vitamin E)
Authors: Leb Laszlo , Beatson Patricia , Fortier Normand , Newburger Peter E , Snyder L Michael
Published: January 2018

The in vitro effect of colloidal suspensions of alpha-tocopherol (α-T) on phorbol- myristate-acetate (PMA)-induced monocyte cytotoxicity and on antibody-dependent monocyte cytotoxicity (ADCC) was studied. We observed that 1) in the presence of α-T, the inhibition was twice as high in the PMA-induced assay than in ADCC; 2) monocytes preincubated with α-T were inhibitory in both assays but much less in ADCC, and 3) target erythrocytes preincubated with a-T decreased the cytotoxicity in the PMA-induced assay only. Since α-T preincubated monocytes showed a decreased release of H2O2 but not of O2″, we concluded that one of the mechanisms by which α-T decreased cytotoxicity could be decreased release of H2O2. Whereas the role of H2O2 was documented in the PMA-induced cytotoxicity, in ADCC non-oxidative injury seems more important. This is supported by 1) lesser inhibition of the assay with α-T preincubated monocytes; 2) lack of protection with α-T preincubated erythrocytes, and 3) mild inhibition with protease inhibitor.
Human Monocyte-Mediated Cytotoxicity Against Herpes Simplex Virus-Infected Cells: Activation of Cytotoxic Monocytes by Free and Liposome-Encapsulated Lymphokines
Authors: Koff Wayne C , Showalter Stephen D , Chakrabarty Mrinal K , Hampar Berge , Ceccorulli Lisa M , Kleinerman Eugenie S
Published: January 2018

Human peripheral blood monocytes were incubated with free or liposome- encapsulated human lymphokines containing macrophage-activating factor (MAF) and tested for their effect on herpes simplex virus (HSV)-infected target cells. Activated monocytes lysed allogeneic HSV type 2 (HSV-2)-infected whole human embryo cells and xenogeneic BALB/c mouse embryo cells (10E2) without any significant effect on uninfected cells, as measured by release of 51Cr from target cells after 18 h of cocultivation. Kinetic studies revealed that lysis of virus-infected cells occurred by 10 h following cocultivation with activated monocytes. The inability of free MAF or supernatants from MAF-acti- vated monocytes to lyse HSV-2-infected cells suggested that direct monocyte- target cell contact is required for monocyte-mediated cytotoxicity of the virus- infected cells. Monocytes activated with MAF suppressed the production of HSV-2 and HSV-1 from virus-infected cells more than control monocytes did. In addition, monocytes treated with liposome-encapsulated MAF selectively destroyed HSV-2-infected cells but left uninfected cells unharmed. The capacity of liposome-encapsulated immunomodulators to activate human monocytes to selectively lyse HSV-2-infected cells has potential therapeutic benefit and should be evaluated in vivo.
Biochemical Characterization of Interleukin 1 From a Human Monocytic Cell Line
Authors: Krakauer Teresa
Published: January 2018

A protein with interleukin 1 (IL-1) activity was obtained from the human acute monocytic leukemia cell line (THP-1) and purified by ultrafiltration, Ultrogel AcA54 chromatography, isoelectric focusing, and discontinuous polyacrylam- ide gel electrophoresis. Like the pi 6.8 species of IL-1 from human peripheral blood monocytes (PBM), the cell line IL-1 has a molecular weight (Mw) of 14,000, a pL of 6.8, is heat labile, and does not bind to concanavalin A- Sepharose. Chemical modification of arginine residues by phenylglyoxal or sulfhydryl groups by N-ethyl maleimide and iodoacetamide completely destroys the activity of IL-1 from THP-1 cells as well as that of the pl 6.8 component from PBM. In contrast, the pi 5.1 component of PBM IL-1 is resistant to heat denaturation and sulfhydryl reagents, although it is totally inactivated by phenylglyoxal. IL-1 from THP-1 cells enhanced the proliferative response of the same subpopulations of PNA- thymocytes as IL-1 from PBM. These observations suggest that IL-1 derived from this cell line is similar to the IL-1 pi 6.8 species produced by human monocytes, and distinct from the pi 5.1 species.
Characteristics of Macrophages in Irradiation Chimeras in Mice Reconstituted With Allogeneic Bone Marrow Cells
Authors: Yasumizu Ryoji , Onoé Kazunori , Iwabuchi Kazuya , Ogasawara Masahiro , Fujita Masaki , Okuyama Harue , Good Robert A , Morikawa Kazuo
Published: January 2018

Biological and immunological characteristics of the reticuloendothelial system of irradiation bone marrow chimeric mice and macrophages collected from various tissue sources of the mice were studied. The chimeras showed comparable activities in carbon clearance to those of normal donor or recipient mice. The macrophages from spleen, lymph node, bone marrow, peripheral blood, liver, peritoneal cavity, and lung were demonstrated to be of donor marrow origin. They showed almost the same enzyme activities and phagocytic capability of sheep erythrocytes (SRBC, E), SRBC sensitized with anti-SRBC IgG (EA), and SRBC sensitized with anti-SRBC IgM and coated with complement (EAC) as those of normal mice. Proportions of Fc receptor and complement receptor-positive cells are also in normal range. In addition, the antigen-presenting capability of the chimeric macrophages for in vitro primary antibody response to SRBC was intact. These observations suggest that the reticuloendothelial system and macrophages of allogeneic bone marrow chimeras where donor and recipient differ at the major histocompatibility complex have no defect so far as could be ascertained by the present study.
Preparation for Hapten Help by Glucan, Muramyl Dipeptide, and Its L-ala-Glycerol-Mycolate Derivative
Authors: Leech Stephen H , Di Luzio Nicholas R , Leclerc Claude
Published: January 2018

Previously, we reported that one of the factors that determines whether or not an animal will be prepared for hapten help after priming is the type of adjuvant used. The present work was undertaken, therefore, to determine which of a diverse variety of adjuvants or biological response modifiers would be effective. They included Freund's complete (CFA) and incomplete (FICA) adjuvants, particulate glucan, muramyl dipeptide (MDP), and its L-ala-glycerol-mycolate derivative. Help by the azobenzenearsonate (ABA) hapten was measured as the augmentation of the anti-bovine γ-globulin (BGG) plaque-forming cell (PFC) response to ABA-BGG of mice that had been hapten-primed with ABA conjugated to ovalbumin (OVA). The results showed that FICA was ineffective. MDP was effective but only if administered with FICA during hapten-priming. MDP-L-ala-glycerol-mycolate was effective without any adjuvant but only within a narrow dose range. Particulate glucan was as effective as CFA in preparing mice for hapten help.

As the macrophage is the primary cellular target of those biological response modifiers that were effective, we conclude that it plays an important role in the cellular interaction involved in the mediation of hapten help.
Effect of Surgical Bursectomy on the Ellipsoid, Ellipsoid-Associated Cells, and Periellipsoid Region of the Chicken's Spleen
Authors: Olah I , Glick B , Taylor R L
Published: January 2018

Surgical bursectomy resulted in cellular depletion of the periellipsoid white pulp, confirming its bursa dependency. Also, in bursectomized birds, the ellipsoid could not be identified, although a small number of abnormal ellipsoid-associated cells (EAC) were observed in the periellipsoid region. The most characteristic finding was the degeneration of the EAC. Degeneration of EAC indicated that the intact bursa was mandatory for normal differentiation of cells of the periellipsoid white pulp into EAC. The promoting effect of the bursa might take place by a bursal hormone. The histological impairment of the EAC was followed by reduced carbon binding and migrating capabilities. Bursectomy resulted in a shift in bacterial phagocytosis in that many cells of the periellipsoid phagocytosed Salmonella. The reduced heterophil infiltration of the ellipsoid in bursectomized birds might be explained by the impaired granular content of the EAC. The impaired migration capability of the EAC might contribute to the low number of germinal centers in bursectomized birds.
Human Recombinant Interleukin-2 Is Mitogenic to Human Lymphocytes
Authors: Mookerjee Basab K , Pauly John L
Published: January 2018

Reported herein are the results of studies demonstrating that purified recombinant human interleukin-2 (hrIL-2) is a potent mitogen for lymphocytes of healthy human donors. The specificity of the hrlL-2-induced response was defined in experiments in which mitogenicity of this T cell growth-promoting lymphokine was completely abrogated by blocking the T cell membrane receptor for IL-2 with the anti-Tac monoclonal antibody. Depletion of adherent mononuclear leukocytes markedly reduced lymphocyte reactivity to hrIL-2, but the response could be fully recovered by the addition of interleukin-1 (IL-1).

Increased proliferative responses were observed using a combination of hrIL-2 and a monoclonal antibody OKT3 that defines a T cell membrane antigen. These studies demonstrate that hrIL-2, as with antigens and phytomitogens, may serve as the first signal of T cell proliferation.
Enhanced Phospholipase A2 and C Activities of Peripheral Blood Polymorphonuclear Leukocytes From Patients With Rheumatoid Arthritis
Authors: Bomalaski John S , Clark Mike A , Douglas Steven D , Zurier Robert B
Published: January 2018

Increased concentrations of eicosanoids are found in synovial fluids (SF) from patients with rheumatoid arthritis (RA). SF polymorphonuclear leukocytes (PMN), which derive from peripheral blood, usually account for approximately 90% of the cells in RA SF. Since eicosanoid precursor fatty acids (FA) are liberated by phospholipases from membrane phospholipids, we examined phospholipase A2 and C activities in peripheral blood PMN from patients with RA. Peripheral blood PMN from patients with RA (RA-PMN) exhibit greater phospholipase A2 activities against phosphatidylcholine (PC) and phosphatidylethanolamine (PE), and greater phospholipase C activities against PC, PE, and phosphatidylinositol (PI) than PMN obtained from normal volunteers (N-PMN).
Dynamics of Arachidonic Acid Metabolism in Macrophages From Delayed-Type Hypersensitivity (Schistosoma mansoni egg) and Foreign-Body-Type Granulomas
Authors: Chensue Stephen W , Ellul Denise A , Spengler Mary , Higashi Gene I , Kunkel Steven L
Published: January 2018

The present study examines the kinetics of arachidonic acid (AA) metabolism by murine macrophages isolated from sites of experimentally induced pulmonary granulomatous inflammation. Macrophages of T-cell-mediated hypersensitivity lesions induced by Schistosoma mansoni eggs (SE-GM) and non-T-cell-mediated foreign-body-type lesions (FB-GM) induced by Sephadex beads were examined. Overall, macrophages from both types of lesions produced mainly lipoxygenase pathway metabolites, leukotrienes, and monohydroxyeicosatetraenoic acids (mono-HETEs). Early after induction (4 days [4D]), SE-GM showed an augmented zymosan-stimulated AA release and metabolism compared to resident peritoneal macrophages. Macrophages from mature lesions (8–32D) showed constitutive synthesis of metabolites and were refractory to zymosan stimulation. Both SE-GM and FB-GM showed augmented AA uptake incorporating a large proportion into neutral lipids. A direct comparison of SE-GM and FB-GM revealed that the T-cell-mediated lesion produced lesser amounts of prostaglandins and leukotrienes and showed reduced incorporation of AA into phosphatidylcholine. These data suggest that AA metabolism by granuloma macrophages is sequentially modified during recruitment and activation at sites of chronic inflammation.
Relationship of Ability of Phospholipids to Stimulate Growth and Bind to Macrophages
Authors: Yui Satoru , Yamazaki Masatoshi
Published: January 2018

Among the phospholipids normally present in mammalian cell membranes, the negatively charged phospholipids, phosphatidylserine (PS), phosphatidylglycerol, and cardiolipin, and the neutral phospholipid phosphatidylethanolamine—but not phosphatidylcholine (PC) or sphingomyelin—were found to induce growth of peripheral macrophages. By use of liposomes prepared from PS, which stimulated growth, and PC, which did not, the relation between growth-stimulating activity and binding of the phospholipids to macrophages was studied. The growth-stimulating activity of PS/PC liposomes decreased with increase in their relative content of PC. The amount of PS bound to macrophages also decreased with increase in the proportion of PC in PS/PC liposomes. These decreases in growth-stimulating and binding activities were both partly recovered by additional incorporation into the PS/PC liposomes of cholesterol, which is also a cell membrane component. Phospholipids that stimulated macrophage growth showed high binding ability to macrophages, whereas those that did not stimulate growth scarcely bound to macrophages. Thus the macrophage growth-stimulating activities of phospholipids correlated well with their ability to bind to macrophages. These liposome models should be useful in elucidating the early mechanism of induction of macrophage growth by lipid materials such as cell debris and lipoproteins.
Tumor-Specific T-Lymphocyte Cytotoxicity Enhanced by Low Dose of C. parvum
Authors: Lau Benjamin H S , Wang-Cheng Rebekah M , Tosk Jeff
Published: January 2018

Three routes of immunotherapy with Corynebacterium parvum (CP) on an ascitic Friend virus-induced leukemia were evaluated. Only the intraperitoneal route, which provided optimal contact between CP and tumor cells, showed prolonged mean survival time. Greatest effectiveness was obtained with multiple injections of CP at weekly intervals and with small initial tumor load. Of particular interest was that lower dosages of CP (5 and 25 μg) gave longer protection than dosages of 50 and 250 μg. Using in vitro 125l-iododeoxyuridine release assay, these lower dosages were shown to selectively enhance the cytotoxicity associated with T lymphocytes, whereas higher dosages appeared to primarily augment the activity of phagocytes. Moderate natural killer cell activity was observed with both the lower and higher dosages of CP. Data from this study indicate that route of administration, dosage of CP, and size of tumor burden are crucial variables determining optimal response to immunotherapy.
Activation of Murine Polymorphonuclear Neutrophils for Fungicidal Activity by Recombinant Gamma Interferon
Authors: Morrison Christine J , Brummer Elmer , Isenberg Richard A , Stevens David A
Published: January 2018

Recombinant murine gamma-interferon (IFN) was tested for its ability to enhance murine polymorphonuclear neutrophil (PMN) fungicidal activity in vitro. PMNs, elicited by intraperitoneal injection of thioglycollate 4 hr prior to collection, were treated with 0.00003-300,000 units of IFN per milliliter for 1 hr prior to challenge with yeast form Blastomyces dermatitidis. These PMNs were not fungicidal for Blastomyces in the absence of IFN; significant enhancement of PMN fungicidal activity by IFN treatment occurred in a dose-dependent manner with maximal enhancement observed at 30,000 U/ml (21% killing). Pretreatment of IFN with rabbit anti-IFN antiserum before addition to PMNs eliminated the enhancement of fungicidal activity by effective doses of IFN. PMN fungicidal activity against phagocytizable Candida albicans was significantly (P < .001) higher (71.3 ± 17.4%) than against 8. dermatitidis. Candidacidal activity was not significantly enhanced by IFN treatment of PMNs. Exogenously added lipopolysaccharide, at levels corresponding to those found in this preparation of IFN, did not activate PMNs for enhanced fungicidal capacity. These data indicate a stimulatory role for IFN in the killing of 6. dermatitidis by PMNs, suggesting that IFN is an active component of the communication between T lymphocytes and PMNs with respect to antimicrobial resistance. They suggest a natural role for IFN in host defense against blastomycosis and other fungal infections, and a possible therapeutic use for exogenous IFN in fungal disease.
Quantitative Microscopy: I. A Computer-Assisted Approach to the Study of Polymorphonuclear Leukocyte (PMN) Chemotaxis
Authors: Cheung Anthony T W , Donovan Richard M , Miller Michael E , Bettendorff Andrea J , Goldstein Elliot
Published: January 2018

A computer-assisted approach has been designed to analyze and quantitate polymorphonuclear leukocyte (PMN) Chemotaxis. This approach involves a rapid, objective, and semiautomated (user-directed) image-analysis system that is video- and microscope-based. The entire system consists of a microvideo set-up that is put on line with a Digital DEC-LSI-11/73 microcomputer, interfaced with a Datacube analog-digital/digital-analog converter. Video signals of PMN movement are digitized by the system at a resolution of 240 pixels vertically by 320 pixels horizontally (at 256 gray levels) and stored in a 76,800-byte frame buffer. The digitized data are stored for later use or utilized immediately for image segmentation, image display, movement, and morphometric computations for each PMN in a maximum phase field (at 645 × high dry) of 50 PMNs at 10-second intervals. The digitized data are used for computation of cell perimeter, surface area, optical density, contour-ratio, position, speed, and direction of locomotion with the utilization of micro-image-analysis programs written in FORTRAN and MACRO assembly language, with the computer operating under RT-11/TSX +. The reliability, objectivity, and reproducibility of measurements made with this quantitative approach have been tested by comparing with manual-tracing measurements of PMN movement. A correlation factor of 0.99 has been obtained. However, the quantitative-microscopic approach is much faster, more objective, less tedious, and much easier to operate than the conventional manual-tracing method.
Cancer Induced Anti-Inflammation and Its Potentiation by Tumor Excision and Rechallenge
Authors: Normann S , Schardt M , Sorkin E
Published: January 2018

Tumor rechallenge following primary tumor excision elicits systemic anti-inflammation that occurs rapidly, affects granulocytes as well as macrophages and is more severe, longer in duration, and induced by fewer tumor cells than the macrophage specific anti-inflammatory effect sometimes seen after primary tumor challenge. Factors important in the pathogenesis of this abnormality are the following. First, primary tumor excision was required as defects did not occur when a second tumor was transplanted during primary tumor growth. Second, the abnormality was restricted to neoplastic cells since normal cells were unable to substitute for either primary or secondary tumor challenge. Third, the anti-inflammatory effect was not due to surgical trauma or local irritation. Fourth, defective inflammation occurred in syngeneic but not allogeneic rats, suggesting an immunological basis for the anti-inflammation. Fifth, elevated glucocorticoids, such as might be expected from an immunological reaction or release of IL-1, may be a contributing but not sole cause for the phenomenon.
Arachidonic and Eicosapentaenoic Acid Metabolism in Bovine Neutrophils and Platelets: Effect of Calcium Ionophore
Authors: Taylor Stephen M , Laegreid William W , Heidel Jerry R , Straub Kenneth M , Liggitt H Denny , Silflow Ron M , Breeze Roger G , Leid R Wes
Published: January 2018

Substitution of dietary fatty acids has potential for altering the inflammatory response. The purpose of the present study was to define the metabolites of arachidonic acid (AA) and eicosapentaenoic acid (EPA) secreted by bovine peripheral blood neutrophils and platelets. High performance liquid chromatography was used to characterize cyclooxygenase and lipoxygenase metabolites secreted in response to the calcium ionophore A23187. Cells were prelabelled with 3H-AA or 3H-EPA prior to challenge with the calcium ionophore. Bovine neutrophils secreted leukotriene B4 (LTB4) and 5-hydroxyeicosatet-raenoic acid (5-HETE) as the major metabolites of AA, as well as the corresponding leukotriene B5 (LTB5) and 5-hydroxyeicosapentaenoic acid (5-HEPE) metabolites of EPA. Peptidoleukotrienes derived from 3H-AA or 3H-EPA were not detected under these conditions. The major tritiated metabolites secreted from bovine platelets were: thromboxane A2, measured as the stable metabolite thromboxane B2 (TXB2); hydroxyhepta-decatrienoic acid (HHT) and 12-HETE derived from 3H-AA; and the omega-3 analogs TXB3 and 12-HEPE, derived from 3H-EPA. Preferred substrate specificities existed amongst the AA- and EPA-derived metabolites for the intermediary enzymes involved in the arachidonic acid cascade. These findings support the hypothesis that substitution of membrane-bound AA by EPA has potential for modulation of the host inflammatory response following cellular phospholipid mobilization.
Generation of Macrophage Variants With 5-Azacytidine: Selection for Mannose Receptor Expression
Authors: Diment Stephanie , Leech Melinda S , Stahl Philip D
Published: January 2018

Mannose receptors are expressed only in primary macrophages. Established macrophage-derived cell lines, although apparently possessing the potential to synthesize mannose receptors, do not express them on their plasma membranes. Using the drug 5-Azacytidine, mannose receptor expression was induced in the macrophage-derived mouse cell line J774. Receptor positive cells were sorted through a fluorescent activated cell sorter (FACS) prior to cloning. Clones were isolated which continuously express mannose receptors in culture. These macrophages were able to endocytose β-glucuronidase and phagocytose yeast particles via mannose receptors. Secretion of the lysosomal enzyme β-hexosaminidase was also reduced in proportion to the degree of mannose receptor expression.
Regulation of Granulocyte Responses in the Blood and Peritoneal Cavity of CBA and B10 Mice During an Acute Inflammation
Authors: Sluiter W , Elzenga-Claasen I , van Hemsbergen-Oomens L W M , van der Voort van der Kley-van Andel A , van Dissel J T , van Furth R
Published: January 2018

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; AUC 0-48 h: 210.9 × 106 granulocytes/mouse × h) and lower in CBA mice AUC0-48 h: 136.8 × 106 granulocytes/mouse × h), whereas the reverse was seen for the granulocyte response in the peripheral blood (AUC0-48 h= 30.5 and 80.7 × 106 granulocytes/mouse × 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.
Eosinophil Granule Cationic Proteins: Major Basic Protein Is Distinct From the Smaller Subunit of Eosinophil Peroxidase
Authors: Weller Peter F , Ackerman Steven J , Smith John A
Published: January 2018

Major basic protein and eosinophil peroxidase are predominant cationic proteins in the cytoplasmic granules of human eosinophilic leukocytes. Each of these proteins has been purified, and major basic protein and the lower molecular weight subunit of eosinophil peroxidase have been found to comigrate on polyacrylamide gel electrophoresis in sodium dodecyl sulfate with similar apparent molecular weights of about 14,700. Because previous molecular weight estimates for these proteins have not recognized the similar molecular weights of these two cationic eosinophil granule constituents, we have evaluated the possible relatedness of these proteins. Upon protein sequence analyses, it was found that the N-terminal 20 amino acid residues of each of these two purified polypeptides differed. These findings established that major basic protein and the smaller subunit of eosinophil peroxidase, although of comparable molecular weights, are two distinct proteins within the cytoplasmic granules of human eosinophils.
Glycolipid Induced Proliferation of Lipopolysaccharide Hyporesponsive C3H/HeJ Splenocytes
Authors: Tomai Mark A , Johnson Arthur G , Ribi Edgar
Published: January 2018

Although the C3H/HeJ mouse is hyporesponsive to lipopolysaccharides (LPS), certain forms of the lipid A fraction have been shown to stimulate cells from this mouse strain. To determine the role of the oligosaccharide chain length on the lipid A-induced proliferation of C3H/HeJ splenocytes, a panel of glycolipids from R-chemotypes (Re, Rc, and Rd) and a nontoxic monophosphoryl lipid A (MPL) were tested. The MPL cells isolated from the MPL of Salmonella minnesota, Salmonella typhlmurium, and the Reglycolipids isolated from Escherichia coli were found to be effective at stimulating the LPS-hyporesponsive spleen cells. A Re-glycolipid isolated from a different strain of E. coli cells was inactive, as were the S. minnesota Rc and Rd chemotypes. Proliferation induced by MPL and the active Re preparations was dose dependent and was inhibited by polymyxin B. Thus, if contamination of the Rc-LPS or MPL with lipid A-associated protein occurred, it was below functional levels. The data suggest that the C3H/HeJ spleen cells are capable of responding to certain glycolipids, but they may lack the ability to convert native LPS into a stimulatory signal. In addition, a monosaccharide precursor of lipid A (lipid X), and a monoacyl glucosamine phospholipid derivative of lipid X (MaGP), were capable of inhibiting the proliferation induced by the MPL and Re-glycolipids. These data are compatible with the existence of a spleen cell receptor for lipid A.
Protein Malnutrition and the Febrile Response in the Fischer Rat
Authors: Bradley Suzanne F , Kauffman Carol A
Published: January 2018

We assessed the effect of protein deprivation on the ability of peritoneal macrophages from Fischer rats to produce interieukin-1 (IL-1) after in vitro stimulation. Pyrogenic activity of supernatants was measured by an in vivo febrile response assay. Control rats were given a 23% casein diet and protein-malnourished rats were given an 8% casein diet for 4 weeks. IL-1-containing supernatants prepared from peritoneal macrophages were injected into assay rats, whose temperatures were measured for 6 hours (δT6). Rats injected with IL-1-containing supernatants derived from peritoneal macrophage cultures of protein-deprived rats had significantly less fever (δT6 = 0.20 ± 0.09°) than rats injected with IL-1 containing supernatants derived from peritoneal macrophage cultures of control rats (δT6 = 0.56 ± 0.09°), P < .01. Protein malnutrition leads to diminished pyrogenicity of macrophage culture supernatants and may be at least partly responsible for the decreased febrile response seen in the malnourished animals.
Diversity of the Human Monocyte/Macrophage System as Detected by Monoclonal Antibodies
Authors: Radzun H J , Kreipe H , Zavazava N , Hansmann M -L , Parwaresch M R
Published: January 2018

Four monoclonal antibodies against the human monocyte/macrophage system, termed KI-M1, KI-M6, KI-M7, and KI-M8, are described with regard to their immunohistochemical tissue distribution pattern and their subcellular reactive sites. The differences found applying these analyses are also reflected by the various molecular weights of the recognized antigens. Based on these data it is proposed that the monocyte/macrophage system can be divided into the phagocytosing compartment on one hand and the immune accessory compartment on the other hand; the latter constitutes the interdigitating reticulum cells, the indeterminate dendritic cells, and the Langerhans cells, as well as the follicular dendritic cells (dendritic reticulum cells) as the accessory cells for T- and B-cell immune response, respectively.
Oxygen Metabolism of the HL-60 Cell Line: Comparison of the Effects of Monocytoid and Neutrophilic Differentiation
Authors: Thompson Blonnie Y , Sivam Geetha , Britigan Bradley E , Rosen Gerald M , Cohen Myron S
Published: January 2018

HL-60 cells are promyelocytic leukemia cells that respond to culture conditions with differentiation into granulocytelike or macrophagelike phagocytes. O2 metabolism is critical to the microbicidal function of phagocytic cells. O2 metabolism was studied in HL-60 cells differentiated with dimethylsulfoxide (Me2SO) and 1,25(OH)2D3, with the objective of 1) determining the validity of these cells as models for human neutrophils and monocytes, respectively, and 2) determining whether these ceils are capable of forming hydroxyl radical. Me2SO-treated cells had morphology consistent with human neutrophils. O2 consumption by these cells in response to phorbol myristate acetate (PMA; 100 ng/ml) or opsonized zymosan (3 mg/ml) was less than that by neutrophils, as was superoxide formation. O2 metabolism was not inhibited by KCN or antimycin A. Myeloperoxidase (MPO) activity decreased during differentiation but remained greater than that of human neutrophils. Cytochalasin B enhanced recovery of superoxide secreted in response to zymosan, implying its release from the phagosome. 1,25(OH)2D3-treated cells had morphology consistent with monocytes. O2 consumption and superoxide release were less than with Me2O-treated cells. Unlike the case with human monocytes, O2 consumption was not inhibited by KCN or antimycin A. MPO activity was minimally reduced by differentiation. Cytochalasin B inhibited recovery of superoxide. Luminol-dependent luminescence was greater among 1,25(OH)2D3-treated cells than among Me2SO-treated cells. Free radicals were also measured with a spin trapping technique using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). Spin trapping allows direct, simultaneous detection of superoxide and hydroxyl radicals. Regardless of the mechanism of differentiation, only superoxide was formed by HL-60 cells. These results show that Me2SO-treated HL-60 cells represent an excellent model for the study of human neutrophil oxidative function. However, 1,25(OH)2D3-treated cells are quite different in their O2 metabolism from peripheral blood monocytes.
Amyloid-Enhancing Factor: Production and Response in Amyloidosis-Susceptible and -Resistant Mouse Strains
Authors: Gervais Francine , Hébert Lise , Skamene Emil
Published: January 2018

Genetic variations in the development of casein-induced amyloidosis exist among inbred strains of mice: CBA/J and C57BL/6J mice are susceptible, while A/J strain mice are resistant to this disease. Amyloidosis is usually induced by daily injections of an inflammatory stimulus for 2-3 wk. The deposition of amyloid in experimental animals can be accelerated by injection of a material called amyloid-enhancing factor (AEF); when injected concomitantly with an inflammatory stimulus, AEF provokes appearance of amyloidosis as early as 2 days after injection. AEF is extracted from amyloid laden or from normal organs (although in small amount).

Our studies were designed to determine if the resistance to amyloidosis seen in A/J mice was either due to a lack of AEF production or to an inability of these mice to respond to AEF. A standard source of CBA/J-derived AEF facilitated the development of amyloidosis in the organs of both the susceptible (CBA/J, C57BL/6J) and the resistant A/J mice. On the contrary, amyloidosis was only induced in susceptible CBA/J hosts when material derived from susceptible (CBA/J, C57BL/6J) animals was injected. CBA/J mice injected with A/J-derived AEF preparation did not develop amyloidosis. These results thus suggest that the determination of resistance or susceptibility to secondary amyloidosis could operate at the level of AEF production.
REVIEW: Methionine Sulfoxide and the Oxidative Regulation of Plasma Proteinase Inhibitors
Authors: Swaim Mark W , Pizzo Salvatore V
Published: January 2018

The sensitivity of methionine residues to oxidation is a mechanism by which many proteins, including plasma proteinase inhibitors, may be oxidatively inactivated. Much evidence suggests that methionine oxidation and concurrent losses of protein activity not only occur widely in living systems but are physiologic, homeostatic processes. Neutrophils, macrophages and other leukocytes secrete large quantities of powerful oxidants at sites of inflammation and may readily bring about methionine oxidative inactivation of proteins. In particular, oxidation of proteinase inhibitors may favorably alter the proteinase-antiproteinase balance to facilitate tissue remodeling and protection from invading organisms. Leukocyte-mediated inhibitor oxidation also appears to regulate local immunosuppressive activity. Pathophysiologic processes such as emphysema and rheumatoidal disease involve derangements of these homeostatic mechanisms.
In Vitro Cytotoxic Activity of Interleukin 2-Dependent Murine Thy-1+ Dendritic Epidermal Cell Lines
Authors: Okamoto Hiroyuki , Itoh Kyogo , Welsh Elizabeth , Trial JoAnn , Platsoucas Chris , Bucana Corazon , Kripke Margaret L
Published: January 2018

Thy-1+ dendritic epidermal cells (Tdec) are bone marrow-derived cells of T-cell lineage. In order to investigate the function of these cells, two interleukin 2-dependent Tdec lines (AU4 and AU16) were established from ear skin of C3H/HeN (MTV-) mice and examined for cytotoxicity against various murine tumor cells. The original phenotype of the Tdec, Thy-1+, AsGM1+, T3+, Lyt-1-, Lyt-2-, Lyt-5+, and L3T4-, was maintained in the cell lines. The Tdec cell lines (AU4 and AU16) lysed a wide spectrum of tumor cell lines in a 4-hr 51Cr-release assay. AU16 was more effective in killing a syngeneic UV-induced skin tumor (UV-2240) and an allogeneic, natural killer cell-resistant tumor (EL-4) than the YAC-1 cell line, which is highly sensitive to natural killer cells. Neither syngeneic nor allogeneic lymphoblasts were killed by AU16. Cytotoxicity was blocked by fixation of AU16 with paraformaldehyde but not by pretreatment of the cells with puromycin or mitomycin C. The finding that two Thy-1+ cell lines derived independently from murine epidermis have unique cytotoxic activity against murine tumor cells suggests that Tdec could play a role in local immune surveillance against skin cancers.
Emergence of a B Lymphocyte Population With ADCC Effector Function in Mammary Tumor Bearing Mice
Authors: Padmanabhan Ranga R , Paul Ronald D , Watson Gordon A , Lopez Diana M
Published: January 2018

Differential expression of antibody dependent cellular cytotoxicity (ADCC) effectors was studied in normal Balb/cCrgl mice and those bearing a chemically induced 7, 12 dimethylbenzanthracene mammary adenocarcinoma. Depletion of macrophages from normal mouse splenocytes by Sephadex G-10 columns resulted in elimination of ADCC. Further separation of the normal G-10 nonadherent splenocytes on nylon wool columns did not result in any population with significant cytotoxicity. However, Balb/c mice bearing mammary tumors showed enhanced levels of ADCC which were not eliminated by macrophage removal. Lymphocytes from tumor bearers further separated on nylon wool yielded nonadherent and adherent populations both capable of effecting significant ADCC. Treatment of the nylon nonadherent cells of both normal and tumor bearing mice with anti-asialo GM1 (AGM1) and complement decreased the ADCC responses. The same treatment only marginally affected cytotoxic levels of nylon adherent cells from tumor bearers, indicating that these effectors are primarily of non-NK lineage. In addition, G-10 nonadherent, nylon adherent cells from tumor bearers separated on a fluorescence activated cell sorter based on the presence of surface immunoglobulins (slg) revealed that both the slg- and slg+ (98% pure) sorted cells were capable of functioning in ADCC. To determine whether in the tumor mice the 2% of slg- cells present in the slg+ sorted population were the ADCC effectors, mixing experiments were done in which up to 10% of slg- cells from tumor bearers were added to nylon adherent cells from normal mice. No significant increases in ADCC levels were found over that of normal mice. These experiments indicate that the 2% slg- cells were not the ADCC effectors nor were they inducing normal B cells to exert this type of cytotoxic reaction in vitro. To further substantiate the B cell lineage of the slg+ ADCC effectors, surface immunoglobulins were removed with protease treatment. After a 36 hr incubation, 92% of the cells had regenerated their slg. The results presented in this paper demonstrate that various splenic lymphoreticular populations from tumor bearers possess an enhanced cytolytic activity against antibody coated target cells. Among these is a unique nylon adherent slg+ cell that is capable of functioning as an ADCC effector.