bluuebunny/crossref_metadata_2025 huggingface dataset:
Metabolism of Glycosylated Human Salivary Amylase: In Vivo Plasma Clearance by Rat Hepatic Endothelial Cells and In Vitro Receptor Mediated Pinocytosis by Rat Macrophages
Authors: Niesen Thomas E , Alpers David H , Stahl Philip D , Rosenblum Jerry L
Published: January 2018

Salivary-type amylase normally comprises about 60% of the amylase activity in human serum, but only a small fraction is a glycosylated isoenzyme (amylase A). In contrast, 1/3 of amylase in human saliva is glycosylated. Since glycosylate can affect circulatory clearance, we studied the clearance of amylase A in rats and its uptake by rat alveolar macrophages.

Following intravenous injection, 125l-labeled amylase A disappeared rapidly from plasma (t 1/2 = 9 min) and accumulated in the liver. Simultaneous injection of mannose-albumin slowed its clearance to a rate comparable to that of 125l-labeled nonglycosylated salivary amylase (t 1/2=45 min). In contrast, galactose-albumin had no effect. Electron microscope autoradiography of the liver following injection of 125l-labeled amylase A revealed a localization of grains over the hepatic endothelial cells.

In vitro studies indicated that amylase A is taken up by alveolar macrophages via receptor-mediated pinocytosis. Uptake was linear over time, saturable, and inhibited by mannan and mannose-albumin, but not by galactose- albumin. We conclude that amylase A, which is a naturally occurring human glycoprotein with at most three terminal L-fucose residues per molecule, is recognized in rats by a mannose receptor located on hepatic endothelial cells. We speculate that this receptor, by rapidly clearing circulating amylase A, may be responsible for the low level of amylase A in human serum.
The Numerous Postulated Biological Manifestations of Interleukin-1
Authors: Kampschmidt Ralph F
Published: January 2018

Over the past 25 years there has been an ever-expanding interest in the mediators that control the immune response [98,133], One of these is produced by macrophages and was first called “lymphocyte activating factor” [49–51] but is now designated “interleukin-1” (IL-1) [118].

As the physical characteristics of IL-1 began to be described, it was noted that they were similar to those of endogenous pyrogen (EP) [128] [151] and leukocytic endogenous mediator (LEM) [85]. Since then, many of the biochemical changes that occur in the host during infection, inflammation, or neoplasia have been attributed to this unique family of monokines. In this review, the family of monokines that has similar physical and biological properties will be referred to as EP/LEM/IL-1. When only one biological activity was measured, the designation will be for that activity. It will be the purpose of this review to examine the evidence that suggests that EP/ LEM/IL-1 has numerous biological activites and to describe its chemical and biological properties, species specificity, and possible sites of action. Finally, I will attempt to address the issue of why there might be a closely related group of mediators with such a broad range of biological activities.
Immunological Activation of Polymorphonuclear Neutrophils for Fungal Killing: Studies With Murine Cells and Blastomyces dermatitidis In Vitro
Authors: Brummer Elmer , Sugar Alan M , Stevens David A
Published: January 2018

The interaction of elicited murine polymorphonuclear neutrophils (PMN) and the thermally dimorphic fungal pathogen Blastomyces dermatitidis in vitro was studied. The PMN elicited intraperitonealy with thioglycollate, in normal mice or mice immune to B dermatitidis, failed to reduce colony forming units (CFU) of B dermatitidis in the inoculum in a 4-hr in vitro assay, even in the presence of 10% fresh immune serum. In contrast, PMN elicited intraperitonealy in immune mice by injection of nonviable B dermatitidis cells significantly reduced inoculum CFU (60 ± 5%) under the same conditions. Furthermore, nonviable B dermatitidis intraperitoneally (i.p.) in normal mice or nonviable Candida albicans i.p. in immune mice failed to elicit peritoneal exudate cells that reduced inoculum CFU in this system. These results support the concept that PMN, elicited in a site by means of an immunological reaction, acquired enhanced microbicidal activity. The fungicidal activity of immunologically elicited PMN was shown to be most effective at high effector to target cell ratios (1,000:1), maximal within 2 hr of coculture, and significantly enhanced in the presence of fresh immune serum compared to heat-inactivated immune serum, normal mouse serum, or fetal bovine serum. Such PMN also had significantly enhanced fungicidal activity against C albicans compared to normal PMN. Fungicidal activity was abrogated in the presence of catalase, implicating hydrogen peroxide generation as the killing mechanism in the activated cells.
Relationships Between Oxidative Metabolism, Macrophage Activation, and Antilisterial Activity
Authors: Godfrey R W , Wilder M S
Published: January 2018

Previous reports from this laboratory have revealed that macrophages obtained from 7-day Listeria-immune mice elicited 15 h before harvest with heat- killed homologous microorganisms were able to kill Listeria monocytogenes while resident or elicited cells were not [14,16]. In the present study, experiments were conducted to determine if phagocytosis-associated oxidative metabolic activity participates in the enhanced destruction of Listeria by activated macrophages. Investigations into production of oxygen radicals by zymosan- stimulated macrophages revealed that Listeria-immune antigen-elicited (LIAE) cells produced significantly more superoxide and hydrogen peroxide than did resident, thioglycolate, or Listeria antigen-elicited macrophages. Additionally, the percentage of nitroblue tetrazolium (NBT) dye postitive cells following exposure to zymosan was maximal in the immune-elicited population. Utilizing a luminol-dependent assay, a short-term chemiluminescent (CL) burst was noted in phagocytizing macrophages. This response was greatest in the LIAE population that exhibited a tenfold increase in peak chemiluminescence over other cell types. Prolonged in vitro culture of these cells diminishes their capacity to generate oxygen radicals. Additionally, LIAE macrophages cultured in excess of 38 h exhibited a significant decrease in zymosan-stimulated hydrogen peroxide release while the decline in superoxide generation was minimal. A substantial diminution in the Listeria-stimulated CL response was also noted during this time period. However, phagocytosis of Listeria by LIAE cells failed to induce the level of oxygen metabolites seen when zymosan was used as the particulate stimulant. In addition, scavengers of oxygen radicals were found to be relatively ineffective in reducing the killing of L monocytogenes by immunologically activated macrophages in culture. It therefore appears that toxic oxygen species do not play a major role in the heightened killing of Listeria by activated macrophages.
Interaction of Plasma Fibronectin (pFN) With Membranous Constituents of Peritoneal Exudate Cells and Pulmonary Macrophages
Authors: Rovin Brad , Molnar Janos , Chevalier Donna , Ng Patrick
Published: January 2018

The prominent role of plasma fibronectin (pFN) in the host defense system as an opsonin for gelatin (collagen)-coated colloids has been established. In the present study we investigated the interaction of pFN and membrane isolates from cells devoid of collagen, as well as several tissues. In a liver slice assay system it was shown that subcellular membrane fractions from lung macrophages, peritoneal exudate cells, spleen, testes, and liver were able to competitively inhibit the pFN-mediated uptake of 125l-gelatin coated latex beads (gLtx*) at low concentrations. Endocytosis of 125l-labeled membrane isolates by macrophage monolayers was also promoted by addition of pFN. In an attempt to characterize the membrane component(s) interacting with pFN, it was found that mild extraction procedure with 1 M KBr could release a significant amount of this inhibitory activity. Further studies demonstrated that the agent(s) responsible for inhibition of gLtx* uptake was heat sensitive, not altered by trypsin treatment, and did not contain actin, a protein known to interact with pFN. This work indicates that pFN interacts specifically with an as yet unknown membrane component(s) and that such interaction will promote clearance of cellular debris by macrophages. This suggests that pFN may be an important opsonin for the reticuloendothelial system in clearance of collagenous and noncollagenous cellular debris once they are exposed to interact with it.
A Model for the Regulation of Myelopoiesis by Specific Factors
Authors: Bender James G , Van Epps Dennis E , Stewart Carleton C
Published: January 2018

Hematopoietic cells originate from stem cells that generate progenitor cells programmed to differentiate along specific cell lineages. Recently, a number of factors have been identified that are involved in regulating the proliferation of myeloid lineage cells. We propose a model in which a hierarchy of specific factors acts sequentially during defined stages of maturation to regulate myelopoiesis. The role of these factors as competence or progression factors in causing cells to enter and traverse the cell cycle is discussed. Experimental evidence supports the model.
Relation of Chemotactic Factor Gradients to Neutrophil Migration and Orientation Under Agarose
Authors: Udén Ann-Mari , Hafström Ingläid , Palmblad Jan
Published: January 2018

Chemotactic substances confer a migratory pattern for neutrohil granulocytes under agarose that is characteristic for each agent. To analyse the cause of such differences, we have studied neutrophil migration and orientation with fMet-Leu-Phe (fMLP), leukotriene B4 (LTB4), and serum as chemoattractants. When these agents were used at optimal concentrations, it was observed that cells stimulated by LTB4 did not start migration as fast and did not migrate as far as those exposed to fMLP, but they maintained a higher degree of orientation. This delay in initiation of migration and maximal degree of orientation was even more marked when serum was the chemoattractant. These migration variables were related to the generation of gradients in the agarose of fML[3H]P, arachidonic-[3H]acid (AA, of which LTB4 is a metabolite), and fluorescein. The curvilinear AA gradient was flatter and more stable than those of fML[3H]P and fluorescein, which were linear. Thus, differences in the development and shape of the gradient of chemoattractant may contribute to differences in migration kinetics.
Studies of Macrophage Function in Murine Systemic Lupus Erythematosus. 3. The Nature, Anatomical Location, and Reversibility of the Phagocytic Defect
Authors: Russell Pamela J , Cameron Fiona H
Published: January 2018

The defect in phagocytosis and binding of antibody-coated sheep erythrocytes (EA) by peritoneal macrophages of (NZB × NZW)F1 or B/W mice is not intrinsic, but is related to the development of the autoimmune disease process. The defect appears to be confined to peritoneal macrophages, since bone marrow (BM)-derived macrophages have normal to elevated activities in vitro. The peritoneal macrophage defect is not due to blockade of Fc receptors in vivo, as shown by long-term culture or recovery of phagocytic and binding activities after removal of Fc receptors by pronase, but represents a reduced number of receptors with slightly delayed turnover. The defect can be reversed by elicitation of activated macrophages with Corynebacterium parvum, thioglycollate, or proteose peptone in vivo. Normal Fc-mediated phagocytosis and binding by BM-derived macrophages cultured from untreated autoimmune mice is enhanced by pretreatment of mice with C. parvum, thioglycollate, or proteose peptone. The cause of the defect in Fc-mediated phagocytosis by resident peritoneal macrophages of autoimmune mice was not ascertained; it may be due to abnormal macrophage kinetics or to the local effects of lymphokines released as a result of other autoimmune changes.
The Significance of Local Resident Pulmonary Alveolar Macrophage Proliferation to Population Renewal
Authors: Sawyer Richard T
Published: January 2018

The local proliferative capacity of resident pulmonary alveolar macrophages (PAM) was determined in normal CD-1 (ICR) mice and mice made severely monocytopenic with strontium-89 (89Sr). Intravenous injection of 2 μCi/gm BW 89Sr rendered mice monocytopenic for a 1-month period. During this time, in addition to there being no change in the size of the PAM population, flash 3HTdR labeling studies showed that at any one instant about 8.8 × 103 of PAM in 88Sr-treated controls and 9.2 × 103 PAM in 89Sr-treated mice were in S phase of the cell cycle. Metaphase arrest experiments using vincristine showed that in 88Sr-treated mice PAM entered M phase at a rate (rM) of 0.51%/hr with a duration of M (Tm) of 0.37 hr, whereas rM = 0.23%/hr with Tm = 0.14 hr for PAM in 89Sr-treated mice. These data suggest PAM population half-times of 131 hr in the presence of monocytes and 301 hr in the absence of monocytes. The data show that PAM proliferate locally; moreover, for intervals longer than the estimated half-time, PAM population size was unchanged. By following the accumulation of pulse 3HTdR-labeled mitotic figures it was found that in both 88Sr- and 89Sr-treated mice G2 = 3 hr and Ts = 11 hr. From these data a model of PAM population growth was used to show that PAM renewal may be accounted for by local proliferation alone. Taken together the data show that resident PAM populations may be sustained by local division of cells and that the influx of peripheral blood monocytes was not needed to sustain PAM.
Chemotactic Activity of Collagen-Like Polypeptides for Human Peripheral Blood Neutrophils
Authors: Laskin Debra L , Kimura Terutoshi , Sakakibara Shumpei , Riley David J , Berg Richard A
Published: January 2018

Damage to interstitial connective tissue is associated with the rapid accumulation of monocytes and neutrophils at the site of injury. To study the role of collagen fragments in neutrophil migration, we analyzed the chemotactic properties of peptide fragments of bovine collagen digested with bacterial collagenase or cyanogen bromide and small molecular weight synthetic polypeptides containing proline (Pro), hydroxyproline (Hyp), and glycine (Gly), the major amino acids that comprise collagen. Using the Boyden chamber and under agarose techniques, we found that collagen fragments were as potent in inducing chemotaxis in neutrophils as the bacterial-derived peptide f-met-leu-phe. The synthetic polytripeptides (Pro-Pro-Gly)5 and (Pro-Hyp-Gly)5 were found to be equipotent in inducing chemotaxis, producing a maximal induction of chemotaxis at 5–10 nM. This suggests that Hyp, the unique imino acid found in collagen, is not required for chemotactic activity. Increasing the length of the synthetic tripeptide from 5 to 10 subunits decreased its chemotactic activity, while the single tripeptide subunit (Pro-Hyp-Gly)1 was the least active peptide, inducing a maximal response at 100 nM. To study the structural requirements for chemotaxis, Pro-Hyp-Gly tripeptides were synthesized with modifications at the N and C terminals ends. Addition of a methyl group to the carboxyl of Gly to form an ester enhanced the chemotactic activity of the peptide by 50%, while substitutions on the amino terminus with an acetyl group decreased the chemotactic activity by 50%. Substitution on the amino terminus with a Boc group decreased the chemotactic activity by 100%. These results indicate that there are specific structural requirements for chemotaxis induced by peptides having a collagen-like sequence of amino acids.
Changes in the Incorporation of Free Fatty Acids Upon the Stimulation of Human Polymorphonuclear Leukocytes
Authors: Phillips Wayne A , Mossmann Horst , Ferber Ernst
Published: January 2018

We have investigated the incorporation of free fatty acids into the cellular lipids of human polymorphonuclear leukocytes (PMN). Resting PMN incorporated both saturated and unsaturated fatty acids into triacylglycerol with only small amounts incorporated into the phospholipids. In contrast, PMN stimulated with the calcium ionophore A23187 incorporated significantly higher amounts of fatty acids, predominantly those other than arachidonic acid, into phosphatidylcholine and phosphatidylinositol, with reduced incorporation into triacylglycerol. Stimulation of PMN with serum-treated zymosan or the chemotactic peptide f-met-leu-phe but not phorbol myristate acetate, also increased the incorporation of fatty acids into these phospholipids. This stimulation-induced incorporation of fatty acids into cellular phospholipids was directed exlusively into position 2 of the lipid and probably reflects the reacylation of lysophospholipids after the release of arachidonic acid by phospholipase A2.
Age Differences in Phagocytosis by Polymorphonuclear Leukocytes Measured by Flow Cytometry
Authors: Nagel James E , Han Kyungeun , Coon Patricia J , Adler William H , Bender Bradley S
Published: January 2018

Elderly persons have increased morbidity and mortality due to bacterial infections. Since the polymorphonuclear leukocyte (PMN) is a major defense against bacterial infection, we utilized fluorescent microspheres and flow cytometry to examine phagocytosis by PMNs from 55 young and middle-aged adults (mean age 41.5 yrs) and two groups of elderly subjects: one group of 35 healthy individuals (mean age 74.1 years) living at home, and a second group of 11 residents (mean age 83.1 years) with severe mental and physical disabilities, living in a domiciliary care facility. We determined the percent phagocytic PMNs, the number of microspheres per PMN, and the number of microspheres per 100 PMNs. The mean number of microspheres per phagocytic PMN was similar for all groups. Statistically significant differences were found between the young and middle-aged group and the healthy or ill elderly groups for the percent phagocytic PMNs (75.3% vs 51.5% and 43.8%), and the number of microspheres per 100 phagocytic PMNs (197.3 vs 131.4 and 103.2). There were no significant differences in these parameters between healthy and debilitated elderly subjects. These data document that there is an age-related increase in representation of a population of PMNs which have a defect in phagocytic ability.
Kinetics of Macrophage Recruitment and Turnover in Peritoneal Inflammatory Exudates Induced by Salmonella or Thioglycollate Broth
Authors: Hopper K E
Published: January 2018

Kinetics of peritoneal macrophage turnover during infection of mice with Salmonella enteritidis or following injection with thioglycollate broth or other peritoneal stimulants has been studied. Single intravenous injections of tritiated thymidine were given and the cells were examined by autoradiography. Maximum labelling of small adherent peritoneal macrophages occurred when 3H-thymidine was given 1 d after Salmonella and the cells were harvested 1 d later. Labelled cells decreased at later times despite maintenance of high numbers of macrophages in the exudates. Results from experiments in which labelled peritoneal cells were reinjected indicated that small, monocyte-enriched, labelled cells were not the major source of the large macrophages. Similar labelling at 2 d was observed using heat-killed Corynebacterium parvum or lipopolysaccharide (LPS) as ip stimulants.

Following injection of thioglycollate broth, labelled peritoneal macrophages were only detectable if 3H-thymidine was given before the stimulant. These labelled cells remained longer in the peritoneal cavity. Labelling of and numbers of blood monocytes were consistent with the promotion of monocytopoiesis by Salmonella but not by thioglycollate. The response to thioglycollate but not Salmonella was dependent on the age of the mice. Animals injected with thioglycollate 1 d before Salmonella also had decreased resistance to bacteria and low numbers of labelled peritoneal macrophages. We propose that thioglycollate may recruit from a subset of preformed monocytes and temporarily block monocytopoiesis or macrophage bactericidal activity.
Measurement of Active Phagocytosis by Polymorphonuclear Leukocytes by Fluorescence Liberation From Phagocytized Microspheres
Authors: Suzuki Kazuo , Uchida Takafumi , Sakatani Tatsuichiro , Sasagawa Sumiko , Hosaka Shuntaro , Fujikura Toshio
Published: January 2018

Phagocytosis by polymorphonuclear leukocytes (PMN) was determined by a newly developed technique based on measurement of liberation of a fluorescence substance from PMN phagosomes; 4-methylumbelliferyl-beta-D-glucuronide (4MUGL), which is a substrate of beta-glucuronidase in lysosome, was conjugated with a microsphere, and 4-methylumbelliferone (4MU) liberated from phagocytized 4MUGL-microspheres was measured. The microspheres were composed of glyceryl-methacrylate having a diameter of 2.0 μm. Liberating activity of six kinds of 4MUGL-microspheres containing various amounts of amino and carboxyl groups was compared. Among these six kinds of 4MUGL-microspheres, four kinds showed activity similar to that of morphological phagocytosis. These four kinds of 4MUGL-microspheres liberated 4MU into the extracellular fluid from PMN during phagocytosis. Furthermore, they were recognized as a substrate of purified beta-glucuronidase. 4MUGL-MS610 showed the highest liberating activity among the four kinds of microspheres. Optimal conditions for phagocytosis by PMN were determined using 4MUGL-MS610. Total liberation of 4MU from the microspheres increased almost linearly with incubation time with PMN from 0 to 60 min and was linear with 4MUGL-MS in concentrations up to 4 × 108 microspheres/ml. This liberation was parallel to phagocytosis in a dose-dependent fashion. During 10-min incubation 20.4% of 4MU was liberated from 4MUGL-microspheres with phagocytosis. Seventy-five percent of the liberated 4MU was distributed in the extracellular fluid. 4MU distributed in the extracellular fluid was not attributable to hydrolysis of unphagocytized microspheres by beta-glucuronidase extracellularly leaked from PMN by phagocytosis. Also phagocytized 4MUGL-MS610 by PMN was observed by scanning electron microscopy.

These results indicate that 4MU was liberated from 4MUGL-MS by hydrolysis due to beta-glucuronidase released into phagosomes with phagocytosis by PMN. Sensitivity of this assay was limited to about 50 pmol/ml, being less than 0.5–1 microsphere phagocytized into one cell.
Oxygen-Reactive Metabolites Are Not Detected at the Effector-Target Interface During Natural Killing
Authors: Storkus Walter J , Dawson Jeffrey R
Published: January 2018

Oxygen-reactive metabolites are not detected at the natural killer (NK)-target cell interface in quantities comparable to those seen for other effector-target cell interactions. A novel luminol-coated target cell chemiluminescence assay is described in which luminol is conjugated to the target cell surface with the bifunctional crosslinker 3,3′-dithiobis(propionic acid N-hydroxysuccinimide ester) (DSP). This modification of the basic chemiluminescence assay precludes exclusion of the detection system from the tightly occluded intercellular junction, a possible deficiency in previous investigations. Luminol conjugation does not affect NK-mediated conjugate formation or cytolysis. As NK activity is enriched by a standard series of effector fractionation procedures, chemiluminescence generated against labeled target cells diminishes. Residual chemiluminescence in the most highly NK-active effector fraction is ablated upon antibody and complement depletion of MO2+ cells. This indicates that monocyte contamination is the source of luminol-detectable oxygen-reactive metabolites.
Effect of Inapparent Murine Hepatitis Virus Infections on Macrophages and Host Resistance
Authors: Dempsey Walla L , Smith Abigail L , Morahan Page S
Published: January 2018

Inapparent infections of mice with murine hepatitis virus (MHV) altered host resistance to experimental infection with a second virus, encephalomyocarditis virus (EMC), reduced the protective effects of exogeneously administered interferon against EMC infections, and it altered macrophage ectoenzyme phenotypes in two macrophage populations. Resident peritoneal macrophages from mice experimentally infected with one of two strains of MHV also demonstrated altered ectoenzyme phenotypes. These data demonstrate that inapparent infections with MHV alter several host resistance and macrophage parameters and directly demonstrate that effects of inapparent MHV infection on macrophage parameters can be reproduced experimentally.
Enhanced Luminol-Dependent Chemiluminescence of Stimulated Rat Alveolar Macrophages by Pretreatment With Alveolar Lining Material
Authors: Webb Deborah S A , Jeska Edward L
Published: January 2018

Previous reports indicate that the in vitro bactericidal activity of rat alveolar macrophages (AM) is dependent on the lipid fraction (ALM-L) of the alveolar lining material (ALM). The present study demonstrates that luminol-dependent chemiluminescence of stimulated rat AM is increased when rat AM are preincubated in the ALM or in the ALM-L. Evidence is presented that oxidation of the unsaturated lipids is responsible for the increase. In addition to pretreatment with the ALM, cells were also pretreated in commercial preparations of several lipids found to be present in the ALM. Preincubation in these lipids produced a significant increase in the luminol-dependent chemiluminescence response. However, when a saturated lipid, dipalmitoyl phosphatidycholine, was used no increase was found. Pretreatment in ALM did not increase the nitro blue tetrazolium dye reduction by the AM, nor was the phagocytosis of latex beads by the AM altered by the addition of the ALM.
Cell-Surface Carbohydrates in Cell Recognition and Response
Authors: Brandley Brian K , Schnaar Ronald L
Published: January 2018

Complex carbohydrates coat the surfaces of cells and have the potential to carry the information necessary for cell–cell recognition. Sugar-specific receptors (lectins) are also present on cells, and can interact with sugars on apposing cells. This may result in the adhesion of the two cells via carbohydrates and specific cell-surface receptors. Such carbohydrate-directed cell adhesion appears to be important in many intercellular activities including infection by bacteria and viruses, communication among cells of lower eukaryotes, specific binding of sperm to egg; and recirculation of lymphocytes, among others [50]. New approaches involving synthesis of chemically defined cell-surface analogs, in conjunction with inhibition experiments, are beginning to reveal the mechanics of a potential carbohydrate “language” involved in intercellular interactions.
Differential Recovery of Antibody Production Potential After Sublethal, Whole-Body Irradiation of Mice
Authors: Brooks Bradford O , Newman Edward L , Reed Norman D
Published: January 2018

Mice were given single injections of sheep erythrocytes (SE) or polyvinylpyrrolidone (PVP) at various times after sublethal, whole-body irradiation (550 rad 60CO) and direct, antigen-specific, plaque-forming cell (PFC) responses were quantified. Irradiated mice did not respond to SE or PVP when immunized 15 d postirradiation (PI); by day 30 PI, the responses by irradiated mice were 40–126% of normal to SE and 3–38% of normal to PVP. The impaired recovery after irradiation of immune responses to PVP was not due to altered antigen dose requirements or altered time of peak PFC response and occurred after irradiation of mice by doses as low as 200 rad. Both athymic and euthymic mice had impaired responses to PVP after whole-body irradiation. The impaired response of irradiated mice to PVP was repaired by adoptive transfer of normal bone marrow, fetal liver, or spleen cells and also by spleen cell preparations enriched in Ig+ cells but not by spleen cell preparations enriched in Thy.1+ or Ig− cells. With the aid of additional antigens it was observed that by day 30 PI, mice had recovered ability to respond to the T-cell-dependent antigen SE and the T-cell-independent type-1 antigens 2,4,6-trinitrophenyl-Brucella abortus and butanol-extracted bacterial lipopolysaccharide, but at that time they gave impaired responses to the T-cell-independent type-2 antigens PVP, type III pneumococcal polysaccharide, and phenol-extracted bacterial lipopolysaccharide; they had an immune response pattern similar to that of CBA/N mice having an X-linked immunodeficiency.
The Ontogeny of Pulmonary Alveolar Macrophages in Parabiotic Mice
Authors: Sawyer Richard T
Published: January 2018

A bilateral distribution of labeled blood monocytes was achieved by unilaterally labeling donor members of surgically parabiosed B6D2F1/J mice with tritiated thymidine (3HTdR). Labeled pulmonary alveolar macrophages (PAM) were found in lung lavage fluid of donors. By contrast, no labeled PAM were found in lung lavage fluid from unstimulated recipients, even 12 days after labeling. Contralateral 3HTdR flash labeling showed that in recipients 3%, or about 104, PAM were in S phase of the cell cycle. Contralateral intratracheal challenge of recipient members with either Corynebacterium parvum vaccine or with Salmonella typhimurium lipopolysaccharide induced the emigration of labeled blood monocytes of donor origin into recipient alveoli. These data show that in parabiotic mice, blood monocytes do not regularly enter the alveolar compartment. It may be concluded that monocytes are not needed on a daily basis to sustain PAM populations.
Eosinophil Chemotactic Factors From Granuloma of Kimura's Disease, With Special Reference to T Lymphocyte-Derived Factors
Authors: Hirashima M , Tashiro K , Sakata K , Muramoto K , Iyama K
Published: January 2018

The granuloma of patients with Kimura's disease characterized by tissue and peripheral blood eosinophilia was reviewed with respect to eosinophil infiltration. An infiltrate of inflammatory cells with histiocytes and a sprinkling of eosinophils were observed in the fibrous stroma surrounding the newly formed vessels. Mast cells were rarely seen in the areas where eosinophils were grouped together. Three different eosinophil chemotactic factors (ECF) were isolated from the granulomas of Kimura's disease. They were termed as low molecular weight (LMW), intermediate molecular weight (IMW), and high molecular weight (HMW)-ECF according to the profile on gel filtration (LMW-ECF, about 500; IMW-ECF, about 12,500; HMW-ECF, 45,000–70,000). In terms of their activity when extracted from the granuloma, LMW-ECF and HMW-ECF seemed to be major natural mediators for the tissue eosinophilia, whereas IMW-ECF was a minor one. In an in vitro system, it was shown that granuloma lymphoid cells produce spontaneously at least two ECF having similar properties to LMW- and HMW-ECF, respectively. By analysis with monoclonal antibodies, granuloma T cells, probably OKT4-positive cells, were shown to be responsible for the production of those two ECF. It was thus suggested that prolonged synthesis of LMW- and HMW-ECF by OKT4-positive T cells plays a crucial role in the local eosinophilia of Kimura's disease.
Fibronectin Is Present on B-Cells but not on OKT 3-Positive T-Lymphocytes or LEU 11-Positive Natural Killer Cells
Authors: Klingemann Hans G , Ebert Jörg , Deeg H Joachim
Published: January 2018

We examined T-cells, B-cells, and natural killer cells of four normal individuals for surface-bound fibronectin (FN) using fluorescence-activated cell sorter analysis and double fluorescence labeling with monoclonal antibodies. While Leu 12 (CD 19)-positive B-cells stained also uniformly with anti-FN antibody, neither OKT 3 (CD 3)-positive T-cells nor Leu 11 (CD 16)-positive natural killer cells could be labeled with anti-FN. Although an anti-FN receptor antibody was not available, these data strongly suggest a distinct pattern of FN-binding by human lymphocytes.
The Immunoregulatory Nature of Iron. II. Lymphocyte Surface Marker Expression
Authors: Bryan Christopher F , Leech Stephen H , Bozelka Brian
Published: January 2018

Previously, we presented preliminary evidence that supported our hypothesis for the immunoregulatory nature of iron [7]. The objective of the present work was to test that hypothesis in greater detail. Our approach was to examine the effect that iron had on the expression of the surface markers on lymphocytes that had been activated by pokeweed mitogen (PWM). The two categories of lymphoid surface molecules were enumerated on those cells; first were those that identify T lymphocytes and second, those that appear on the membrane of T cells following activation. The results, as regards T cell-associated molecules, demonstrated that iron suppresses the expression of the molecules identified by the monoclonal antibodies OKT3 and OKT4. It suppressed expression of the T4 molecule in PWM-activated cells (30.6% ± 4.5; n = 5) compared with untreated but activated cells (52.2% ± 2.9; n = 5; P = 1.9 × 10−3) resulting in a reduced helper:suppressor T cell ratio from 2.2 ± 0.4 to 1.2 ± 0.3. With regard to activation-associated lymphocyte markers, iron significantly enhanced expression of the receptor for transferrin as identified by the monoclonal antibody, OKT9. However, it failed to change significantly the expression of three other activation-associated markers, namely, la, T10, and the receptor that forms thermostable erythrocyte-rosettes (TE-R) with sheep red blood cells (SRC). We conclude from those results that iron has a differential immunoregulatory influence on the expression of certain lymphocyte surface molecules on actively dividing lymphocytes.
Transient Catecholamine Modulation of Neutrophil Activation: Kinetic and Intracellular Aspects of Isoproterenol Action
Authors: Tecoma Evelyn S , Motulsky Harvey J , Traynor Alexis E , Omann Geneva M , Muller Heinz , Sklar Larry A
Published: January 2018

Modulation of neutrophil activation by catecholamines may reflect regulatory mechanisms that couple beta-adrenergic and N-formyl peptide receptors to antagonistic biochemical pathways. We examined kinetic and mechanistic aspects of the inhibition by catecholamines of neutrophil activation by formyl peptides. Inhibition of oxidant production by isoproterenol (ISO) was detected as low as 3 nM, had an ID50 of 10−7 M, and could be blocked and reversed by propranolol. Recovery of cell function occurred over a period of minutes when the concentration of ISO was less than 10−6 M. These observations are discussed in terms of the interaction of ISO with the adrenergic receptors.

The site of catecholamine action is addressed. ISO neither influences formyl peptide-receptor interaction nor does it inhibit oxidant production by phorbol ester. These results suggest an impairment of intracellular signalling processes that couple the formyl peptide-receptor binding to cell activation. We observed inhibition of intracellular Ca+ + elevation by ISO only at low formyl peptide concentrations. This inhibition is consistent with a partial inhibition of phosphoinositide metabolism, which was observed. Several other cell responses, including actin polymerization and right angle light scatter, are minimally inhibited by 10−6 M ISO indicating that the cell activation process is not entirely obliterated. The presence of catecholamine and formyl peptide results in a synergistic elevation of cAMP. The intracellular targets of ISO action may be regulated by cAMP dependent kinases and could follow a branchpoint in the activation sequence that leads distinctly to oxidase activation and cytoskeletal activation.
Naturally Acquired Leprosy in the Nine-Banded Armadillo: A Decade of Experience 1975–1985
Authors: Walsh Gerald P , Meyers Wayne M , Binford Chapman H
Published: January 2018

A decade has passed since our first report of naturally acquired leprosy in the nine-banded armadillo. Our studies and those of others during this period confirm the identification of the etiologic agent as Mycobacterium leprae. Confirmation is based on the results of histopathologic examination and microbiologic evaluations that included attempts to culture the organism, flourescent antibody studies, mycolic acid analysis, and DNA determinations demonstrating complete relatedness between the natural agent and M. leprae. Surveys involving large numbers of animals demonstrate a significant prevalence of the disease in armadillos captured in Louisiana and Texas. The discovery of naturally acquired leprosy in a chimpanzee in 1977 and a sooty mangabey monkey in 1979 reinforce the concept of leprosy as a zoonosis. Extensive contact with armadillos has been implicated by other observers in seven patients with leprosy in Texas. We believe the prevalence of leprosy in wild armadillos requires that they be considered a source of infection in patients from geographic areas where leprosy and armadillos co-exist.
Intravascular Macrophages in the Sheep Lung
Authors: Wheeldon Eric B , Hansen-Flaschen John H
Published: January 2018

The presence of a resident population of intravascular macrophages in the sheep lung was investigated. Injection of fluorescent microspheres into the sheep results in uptake by a population of large phagocytic cells scattered within the alveolar capillaries of the lung. By contrast, the rat lung has no uptake of these fluorescent microspheres into the alveolar walls. The cells are large, extremely irregular in shape, often exceed 30 μm in length, and lie within the vascular lumen, apposed to the underlying endothelium. They have several features characteristic of mature macrophages including numerous pseudopodia, a partially preserved cell coat, wormlike bodies (“micropinocytosis vermiformis”), and numerous phagosomes that often contain erythrocytes. Unlike other species such as the rat, in which the mononuclear phagocyte system is distributed primarily in the liver and spleen, the sheep lung has a significant clearance function. Moreover, the presence of these cells in the lung may have important implications for studies of the inflammatory response to pulmonary vascular injury in the sheep.
Effect of Pentoxifylline on the Phagocytic Activity, cAMP Levels, and Superoxide Anion Production by Monocytes and Polymorphonuclear Cells
Authors: Bessler H , Gilgal R , Djaldetti M , Zahavi I
Published: January 2018

The effect of pentoxifylline (Trental) on the phagocytic capacity, cAMP levels, and superoxide anion production of human peripheral blood monocytes and polymorphonuclears (PMNs) was studied. The drug inhibited the phagocytosis of latex particles by both monocytes and PMNs in a dose-dependent manner. In addition, superoxide anion production during the phagocytic process was also reduced following incubation of the cells with pentoxifylline. It is suggested that this inhibitory effect is due to the increased intracellular levels of cAMP induced by the drug.
Model for Leukocyte Regulation by Chemoattractant Receptors: Roles of a Guanine Nucleotide Regulatory Protein and Polyphosphoinositide Metabolism
Authors: Snyderman Ralph , Smith Charles D , Verghese Margrith W
Published: January 2018

Binding of chemoattractants to their receptors on phagocytes activates a guanine nucleotide regulatory (N) protein through the substitution of GTP for GDP on N. The activated N protein in turn stimulates a membrane-associated phospholipase C by lowering the Ca2+ concentration required to activate this enzyme from supraphysiologic levels to ambient intracellular concentrations. The phospholipase C hydrolyzes phosphatidylinositol 4,5-bisphosphate into the Ca2+ mobilizer inositol 1,4,5-trisphosphate and the protein kinase C activator 1,2-diacylglycerol. In addition to promoting cellular activation, the products of this hydrolysis initiate processes which feed back to inhibit polyphosphoinositide breakdown. The regulatory model proposed herein may be relevant to other receptors which stimulate polyphosphoinositide metabolism.
Preferential Loss of DNA Polymerase α Following Suppression of Replicative DNA Synthesis of Guinea Pig Macrophages by the Immunostimulants Muramyl Dipeptide or Lipopolysaccharide
Authors: Haraguchi Tokuko , Nagao Shigeki , Tanaka Atsushi , Nagano Hiroshi
Published: January 2018

Oil-induced guinea pig peritoneal exudate macrophages were found to incorporate 3H-thymidine into trichloroacetic acid-insoluble fraction. In pulie labeling experiments, the moorporsted 3H-thymiome was detected in short fragmetns or DNVA, wnicn corresponded to the Okazaki fragments. These results indicate that the observed thymidine incorporation is due to nuclear DNA replication but not DNA repair. The observed DNA synthesis of the macrophages was remarkably suppressed when the cells were cuftured in a presence of muramyl dipeptide (MDP) or bacterial lipoporysaccharide (LPS). The significant decrease of DNA polymerase a activity was found in the celts treated with MDP or LPS. in contrast, the activity of polymerase β was not at all affected by the same treatment.
FACS Analysis of Tumor-Associated Macrophage Replication: Differences Between Metastatic and Nonmetastatic Murine Mammary Tumors
Authors: Mahoney Keith H , Heppner Gloria H
Published: January 2018

The proliferation activity of adherent, tumor-associated macrophages (TAM) from three related murine mammary carcinoma lines was measured by fluorescence-activated cell sorter (FACS) analysis of the incorporation of 5′-Bromo-2′deoxyuridine (BrdU) in vivo during a 1 hr period prior to tumor removal. The tumor lines examined were line 67 which is nonmetastatic following either subcutaneous (SC) or intravenous (IV) injection, line 168 which colonizes the lung after IV injection but not SC, and line 66 which can colonize the lung from either site.

The percentage of total cells identified morphologically as macrophages were similar for tumors from lines 66 and 67 (41% and 38%, respectively), as compared to line 168 tumors, in which, after digestion, 27% of cells were identified as macrophages (P < 0.05).

Adherent TAM from line 66 tumors had the greatest percentage of BrdU incorporating cells, with an average of 22%. This was statistically significant (P < 0.01) from TAM from tumors of line 67, which were 13% positive. The percentage of adherent TAM from line 168 tumors (18% positive) was also significantly different from 67 TAM (P < 0.01).

These results demonstrate a possible correlation between the percentage of TAM undergoing replication and the ability of the host tumor to colonize the lung. There is no apparent relationship between the percentage of TAM in replication and the number of macrophages associated with a tumor.
A Role for the Laminin Receptor in Leukocyte Chemotaxis
Authors: Bryant G , Rao C N , Brentani M , Martins W , Lopes J D , Martin S E , Liotta L A , Schiffmann E
Published: January 2018

Previous studies on the mechanism of leukocyte traversal of basement membranes showed that rabbit peritoneal exudate polymorphonuclear cells (PMN) preferentially used laminin, a major constituent of basement membrane, to attach to another component, type IV collagen. PMN also responded chemotactically to nanomolar levels of laminin. We have now determined that PMN possess a receptor for laminin. Scatchard analysis using 125I laminin indicates a single class of saturable high affinity binding sites (kd = 6.15 nM/L) on PMN and 3.6 x 104 sites per cell. A chymotryptically derived fragment of laminin, C1, which lacks both the long arm and the globular end regions of the short arm (i.e., matrix binding sites) but retains the laminin receptor binding region, gave similar results. Immunoperoxidase studies using monoclonal antibodies to the laminin receptor (mAbLR) indicated the presence of the receptor on the surface of PMN. These cells responded chemotactically to nanomolar levels of C1 and laminin, a result consonant with binding data. PMN chemotaxis to a formyl peptide was markedly inhibited by mAbLR, suggesting that the laminin receptor may be required for PMN chemotaxis in general. Our results suggest that PMN extravasation across basement membranes is aided both by reversible attachment of the cells to laminin in the matrix and by chemotaxis to a gradient of soluble intact and possibly degraded laminin. These characteristics have much in common with those of highly metastatic tumor cells.
Monokines Mediate Decreased Hepatic Glucocorticoid Binding in Endotoxemia
Authors: Hill Molly R , Stith Rex D , McCallum Roderick E
Published: January 2018

The purpose of this study was to determine whether endotoxin decreased hepatic glucocorticoid binding by the action of mediator(s). Steroid binding in liver cytosol, plasma glucose levels, and plasma corticosterone levels were assayed in C3HeB/FeJ LPS normoresponsive and C3H/HeJ LPS hyporesponsive mice. In C3HeB/FeJ mice, endotoxin significantly depressed the maximum number of steroid binding sites (Bmax) to 30% of control. Plasma glucose levels were decreased to 50% of control, and plasma corticosterone levels increased 4-fold. No changes in these parameters were seen in C3H/HeJ mice given endotoxin, except for decreased plasma glucose levels at the highest dose of endotoxin. Decreased steroid binding was observed in C3H/HeJ mice 4-6 hours after receiving C3HeB/FeJ peritoneal exudate cells (elicited with thiogiycollate) and endotoxin. No change in steroid binding was observed in C3H/HeJ mice that received C3H/HeJ peritoneal exudate cells and endotoxin. Mediator-rich plasma was produced in CF-1 mice by infecting them with 1 x 107 BCG and by challenging them with endotoxin (2 μg) 2 weeks later for 2 h. Transfer of BCG-endotoxin plasma to C3H/HeJ mice also resulted in decreased steroid binding and plasma glucose. These results indicate that perturbation of glucocorticoid action during endotoxin shock is mediated by soluble factor(s) other than endotoxin. A likely source of mediator(s) is the mononuclear phagocyte.
On the Regulation of Differentiation and Function of Rat Macrophages by 1,25-Dihydroxyvitamin D3 and Dexamethasone
Authors: Shezen Elias , Goldman Rachel
Published: January 2018

Rat bone marrow macrophage progenitor cells develop in vitro in the presence of rat embryo fibroblast conditioned medium into colonies and clusters. 1α,25-Dihydroxyvitamin D3 (1,25(OH)2D3) (0.12-12 nM) was found to enhance the formation of macrophage colonies and the proliferation of mononuclear phagocytes in liquid cultures of bone marrow cells (ED50 0.12-1.0 nM). Fractionation of bone marrow cells by centrifugal elutriation showed that: a) macrophage progenitors are heterogeneous in size; b) the progenitors eluted at early fractions have a lower proliferative capacity (form mainly small clusters) than those eluting at later fractions (higher counterflow velocities) which develop into macrophage colonies and c) that 1,25(OH)2D3 (at 12 nM) augments the expression of colony forming cells enriched in late eluting fractions while having a suppressive effect on expression of low proliferative potential cluster forming cells enriched in early eluting fractions. Dexamethasone was found to suppress the clonal growth of macrophage progenitor cells as well as their proliferation in liquid cultures (ED50 about 1 nM). Both dexamethasone and 1,25(OH)2D3 induced in mononuclear phagocytes of 4 d cultures an increased phagocytic capability. The data suggest a regulatory role for 1,25(OH)2D3 and glucocorticosteroids in myelopoietic processes in the rat. Furthermore, when compared with our recent findings with mouse bone marrow cells, the effects, their magnitude and concentration dependence imply genuine species differences in the responses of mice and rats to these hormones.