Driving Influences of the Doppler Flash Observed by SuperDARN HF Radars in Response to Solar Flares.

Sudden enhancement in high-frequency absorption is a well-known impact of solar flare-driven Short-Wave Fadeout (SWF). Less understood, is a perturbation of the radio wave frequency as it traverses the ionosphere in the early stages of SWF, also known as the Doppler flash. Investigations have suggested two possible sources that might contribute to it's manifestation: first, enhancements of plasma density in the D-and lower E-regions; second, the lowering of the F-region reflection point. Our recent work investigated a solar flare event using first principles modeling and Super Dual Auroral Radar Network (SuperDARN) HF radar observations and found that change in the F-region refractive index is the primary driver of the Doppler flash. This study analyzes multiple solar flare events observed across different SuperDARN HF radars to determine how flare characteristics, properties of the traveling radio wave, and geophysical conditions impact the Doppler flash. In addition, we use incoherent scatter radar data and first-principles modeling to investigate physical mechanisms that drive the lowering of the F-region reflection points. We found, (a) on average, the change in E- and F-region refractive index is the primary driver of the Doppler flash, (b) solar zenith angle, ray's elevation angle, operating frequency, and location of the solar flare on the solar disk can alter the ionospheric regions of maximum contribution to the Doppler flash, (c) increased ionospheric Hall and Pedersen conductance causes a reduction of the daytime eastward electric field, and consequently reduces the vertical ion-drift in the lower and middle latitude ionosphere, which results in lowering of the F-region ray reflection point.

Community-Based Recreation Therapy and Mental Health Recovery: A Mixed-Media Participatory Action Research Study.

BACKGROUND: Personal recovery is an individualized process through which people develop a positive identity and live a meaningful life, with symptoms of mental illness. Few studies have explored the role of recreation therapy in the recovery process from the perspectives of individuals with lived experience of mental illness. OBJECTIVES: To understand how community-based recreation therapy can support mental health recovery, from the perspectives of people diagnosed with mental illness, and to guide the development, delivery and evaluation of recovery-oriented mental health services. METHODS: Guided by the principles of participatory action research (PAR) and photovoice, six participant researchers (PRs) generated arts-based media and narrative data in response to the research question: How can therapeutic recreation, in a community mental health center, support the recovery of individuals diagnosed with mental illness? The research group analyzed the qualitative data through a participatory data analysis process. RESULTS: The PRs produced and analyzed 24 pieces of arts-based media and 5 hours of transcribed narrative data describing their artworks' relationship to therapeutic recreation and recovery. The PRs identified seven salient themes through the participatory data analysis process: providing a safe place, promoting hope, finding balance, developing self-wisdom, increasing enjoyment, building confidence, and encouraging self-determination. CONCLUSIONS: Service recipients' unique preferences and perspectives must be integral to service development to deliver therapeutic recreation interventions that are truly recovery-oriented. The research findings will be used to guide the delivery of innovative, collaborative, person-centered programming in community-based mental health settings.

Long-lasting poloidal ULF waves observed by multiple satellites and high-latitude SuperDARN radars.

Poloidal ultra-low frequency (ULF) waves between 5-10 mHz were observed by multiple satellites and three high-latitude Super Dual Auroral Radar Network (SuperDARN) radars during the recovery phase of a moderate geomagnetic storm on Jan 24-27, 2016. The long-lasting ULF waves were observed in the magnetic field and energetic particle flux perturbations during three successive passes by two Geostationary Operational Environmental Satellites (GOES) through the dayside magnetosphere, during which plasmasphere expansion and refilling were observed by two Time History of Events and Macroscale Interactions during Substorms (THEMIS) probes. The radial magnetic field oscillation was in phase (~ 180° out of phase) with the northward (southward) moving proton flux oscillation at 95 keV, consistent with high-energy drift-bounce resonance signatures of protons with second harmonic poloidal standing Alfvén waves. The longitudinal extent of the waves approached 10 hours in local time on the dayside and gradually decreased with time. High-time resolution (~ 6 s) data from three high-latitude SuperDARN radars show that the wave intensification region was localized in latitude with a radial extent of ~ 135-225 km in the subauroral ionosphere. No signature of these waves were observed by ground-based magnetometers colocated with the GOES satellites suggesting that the poloidal waves were high-m mode and thus screened by the ionosphere. During this interval one of the THEMIS probes observed a bump-on-tail ion distribution at 1-3 keV which we suggest is the source of the long-lasting second harmonic poloidal ULF waves.

Survey of ionospheric Pc3-5 ULF wave signatures in SuperDARN high time resolution data.

Ionospheric signatures of ultra-low frequency (ULF) wave in the Pc3-5 band (1.7-40.0 mHz) were surveyed using ~6 s resolution data from Super Dual Auroral Radar Network (SuperDARN) radars in the northern hemisphere from 2010 to 2016. Numerical experiments were conducted to derive wave period dependent thresholds for automated detection of ULF waves using the Lomb-Scargle periodogram technique. The spatial occurrence distribution, frequency characteristics, seasonal effects, solar wind condition and geomagnetic activity level dependence have been studied. Pc5 wave events were found to dominate at high and polar latitudes with a most probable frequency of 2.08 ± 0.07 mHz while Pc3-4 waves were relatively more common at midlatitudes on the nightside with a most probable frequency of 11.39 ± 0.14 mHz. At high latitudes, the occurrence rate of Pc4-5 waves maximizes in the dusk sector and during winter. These events tend to occur during low geomagnetic activity and northward interplanetary magnetic field (IMF). For the category of radially bounded but longitudinally extended Pc4 events in the duskside ionosphere, an internal driving source is suggested. At midlatitudes, the Pc3-4 occurrence rate maximizes premidnight and during equinox. This tendency becomes more prominent with increasing auroral electrojet (AE) index and during southward IMF, which suggests many of these events are Pi2 and Pc3-4 pulsations associated with magnetotail dynamics during active geomagnetic intervals. The overall occurrence rate of Pc3-5 wave events is lowest in summer, which suggests that the ionospheric conductivity plays a role in controlling ULF wave occurrence.

Tumour gene expression predicts response to cetuximab in patients with KRAS wild-type metastatic colorectal cancer.

BACKGROUND: Although it is accepted that metastatic colorectal cancers (mCRCs) that carry activating mutations in KRAS are unresponsive to anti-epidermal growth factor receptor (EGFR) monoclonal antibodies, a significant fraction of KRAS wild-type (wt) mCRCs are also unresponsive to anti-EGFR therapy. Genes encoding EGFR ligands amphiregulin (AREG) and epiregulin (EREG) are promising gene expression-based markers but have not been incorporated into a test to dichotomise KRAS wt mCRC patients with respect to sensitivity to anti-EGFR treatment. METHODS: We used RT-PCR to test 110 candidate gene expression markers in primary tumours from 144 KRAS wt mCRC patients who received monotherapy with the anti-EGFR antibody cetuximab. Results were correlated with multiple clinical endpoints: disease control, objective response, and progression-free survival (PFS). RESULTS: Expression of many of the tested candidate genes, including EREG and AREG, strongly associate with all clinical endpoints. Using multivariate analysis with two-layer five-fold cross-validation, we constructed a four-gene predictive classifier. Strikingly, patients below the classifier cutpoint had PFS and disease control rates similar to those of patients with KRAS mutant mCRC. CONCLUSION: Gene expression appears to identify KRAS wt mCRC patients who receive little benefit from cetuximab. It will be important to test this model in an independent validation study.

Agricultural dust production in standard and conservation tillage systems in the San Joaquin Valley.

The negative health effects of repeated dust exposure have been well documented. In California's San Joaquin Valley, agricultural operations may contribute substantially to airborne particulates. We evaluated four management systems to assess impacts on dust production and soil properties for a cotton (Gossypium hirsutum L.)-tomato (Lycopersicon esculentum Mill.) rotation: standard tillage with (STCC) and without (STNO) cover crop, and conservation tillage with (CTCC) and without (CTNO) cover crop. Gravimetric analysis of total dust (TD, <100-mum aerodynamic diameter) and respirable dust (RD, 4-mum aerodynamic diameter) samples collected in the plume generated by field implements showed that dust concentrations for CTNO treatments were about one-third of their STNO counterparts for both cumulative TD and RD measured throughout the two-year rotation, primarily due to fewer in-field operations. The TD and RD production for STNO and STCC was comparable, whereas the CTCC system produced about twice as much TD and RD as CTNO. Energy dispersive spectroscopy (EDS) analyses showed absolute increases of 8 and 39% organic fragments in STCC and CTCC over STNO and CTNO, respectively, while organic fragments in the TD increased by 6% in both cover crop treatments. Soil C content was positively correlated with clay content and increased by an average of 0.12 and 0.07% in the cover crop and non-cover crop treatments, respectively, although soil C for each treatment showed a distinct response to a field texture gradient. While dust emissions show an immediate decrease due to fewer field operations for the conservation tillage treatments, long-term sampling is necessary to determine the effects that increased aggregation through organic matter additions may have on dust production.

Type and crossmatch of the trauma patient.

OBJECTIVE: To identify a population of trauma patients in the emergency department (ED) that do not require emergent blood transfusion via a combination of clinical risk factors that are readily accessible and easily obtained. METHOD: A review of trauma patients was conducted for a 6-month period. Crossmatched patients were identified and examined for clinical characteristics and whether transfusion was performed. Risk factors for transfusion were identified and a model was developed for predicting likelihood of transfusion. RESULTS: Six hundred fifty-four patients were crossmatched, with emergent transfusion occurring in 81 (12.4%). Four risk factors were identified: systolic blood pressure < 90 mm Hg, Glasgow Coma Scale score < 9, pulse > 120 beats/min, and high-risk injury (trauma to the chest between the midclavicular lines, abdominal injury with diffuse tenderness, survival of a fatal vehicular crash, ejection from a vehicle, or stab or gunshot wound to the trunk). Patients with no risk factors were shown to have a 2.2% incidence of transfusion with no emergent transfusions occurring in the ED. CONCLUSION: Trauma patients with no risk factors at presentation were less likely to require emergent blood transfusion, especially in the setting of the ED.

Protection against endotoxic shock by bactericidal/permeability-increasing protein in rats.

Bactericidal/permeability-increasing protein (BPI) is a neutrophil primary granule protein that inhibits effects of LPS in vitro. The current study examined the effects of BPI on hemodynamics, mortality, and circulating endotoxin and cytokines in conscious rats with endotoxic shock. Catheters were implanted into the right femoral artery and vein. 1 d later, human recombinant BPI (10 mg/kg) or vehicle was intravenously injected immediately, 30 min, or 2 h after intravenous injection of LPS (7.5 mg/kg). Mean arterial pressure (MAP) and heart rate were monitored and blood was collected before and after injection. BPI given immediately or 30 min after LPS prevented the LPS-induced reduction in MAP at 4-8 h and markedly reduced mortality. BPI given 2 h after LPS injection had no protective effect. BPI treated immediately after LPS reduced the circulating levels of endotoxin and IL-6 but increased the circulating levels of TNF. We propose that BPI exerts its protective effect through a TNF-independent mechanism, by inhibiting endotoxin-stimulated production of IL-6.

Intravenous interleukin-8 inhibits granulocyte emigration from rabbit mesenteric venules without altering L-selectin expression or leukocyte rolling.

Injection (i.v.) of the granulocyte chemoattractant/activator IL-8 has been shown to reduce neutrophil recruitment into dermal inflammatory sites in vivo. To further investigate the mechanism of this phenomenon, we examined the effect of i.v. [Ser-IL-8]72 (12-20 micrograms/kg) on leukocyte rolling and chemoattractant-induced emigration in mesenteric venules of New Zealand White rabbits and on expression of L-selectin (mAb LAM1-3) and CD18 (mAb 60.3) on circulating rabbit granulocytes. Within 1 min of IL-8 i.v., granulocytes virtually disappeared from carotid blood samples for approximately 5 min. Concomitantly, the flux of rolling leukocytes in mesenteric venules fell from 83 +/- 21 to 2 +/- 1 leukocytes/min. Both rolling leukocyte flux and systemic granulocyte count returned to or exceeded control values within less than 30 min. The chemoattractant/activator FMLP (0.15 microgram/kg i.v.) produced similar results. A second i.v. injection of IL-8 or FMLP, 90 min after the first challenge, had equipotent effects. Local extravascular application of IL-8 via micropipette close to a venule induced adhesion and emigration of 63 +/- 21 leukocytes per site before, but only 26 +/- 9 leukocytes per site 50 to 75 min after i.v. IL-8, when systemic granulocyte count and rolling leukocyte flux had reached or exceeded control values. This was not due to agonist-specific desensitization, because a similar reduction of leukocyte emigration was seen after FMLP i.v. Rabbit granulocytes circulating in vivo uniformly expressed near-control levels of L-selectin at all times between 3 and 360 min after IL-8 i.v. CD18 expression transiently increased after IL-8 i.v. and returned to base line by 90 min. These findings show that IL-8 i.v. reduces granulocyte recruitment to inflammatory sites by inhibiting function(s) necessary for transmigration that are independent of L-selectin and subsequent to rolling.

Isolation and characterization of an intracellular serine proteinase inhibitor from a monkey kidney epithelial cell line.

A recent report described a thrombin inhibitory activity in the soluble fraction of human placenta and the cytosolic fraction of K562 cells. Isolation and characterization of the functionally inactive 35-38-kDa placental form of this protein revealed that it was a novel serine proteinase inhibitor (Coughlin, P. B., Tetaz, T., and Salem, H. H. (1993) J. Biol. Chem. 268, 9541-9547). In the present study, we observed a 67-kDa sodium dodecyl sulfate (SDS)-stable complex when 125I-thrombin was incubated with the cytosolic fraction of a monkey kidney epithelial cell line, BSC-1. This complex was not observed in either the particulate cell fraction extracted with 0.2% Triton X-100 or medium conditioned by cells, suggesting that the thrombin-complexing factor is confined to the cytoplasm. The cytoplasmic antithrombin activity was purified to apparent homogeneity from the cytosol of BSC-1 cells previously pulsed with [35S]methionine by a combination of heparin-agarose chromatography, Mono Q fast protein liquid chromatography, and anhydrotrypsin-Affi-Gel 10 affinity chromatography. Analysis of the affinity-purified preparation by SDS-polyacrylamide gel electrophoresis and fluorography revealed a single protein with an apparent molecular mass of 38 kDa. The purified 38-kDa protein inhibited the amidolytic activities of thrombin, trypsin, urokinase, and factor Xa but not that of elastase. Incubation of the 38-kDa protein with excess thrombin identified approximately 60% of the labeled 38-kDa protein in an SDS-stable 67-kDa complex. The purified 38-kDa inhibitor was cleaved with cyanogen bromide and the isolated peptides subjected to microsequencing. Amino acid sequence obtained for a region within this protein exhibited significant homology with human antithrombin III and plasminogen activator inhibitors 1 and 2. This homologous peptide contained the full complement of residues designated as highly conserved in helix F of the greater serine proteinase inhibitor superfamily. In addition, an internal sequence of GGGGDIHQGF was found in the monkey cytoplasmic inhibitor, which is identical to that reported for an internal sequence of the human placental inhibitor. These findings confirm the existence of a novel cytoplasmic serine proteinase inhibitor in mammalian cells and provide additional details of its molecular properties. The physiological function of this novel serine proteinase inhibitor in cytoplasm is unknown.

Protease nexin 1 is expressed in the human placenta.

Protease nexin 1 (PN1), a serine protease inhibitor that inactivates thrombin, urokinase, and plasmin, is produced abundantly in cultures of human fibroblasts and rat and human glioma cells. The major sites of PN1 synthesis in vivo and the specific physiological function(s) of this serpin are unknown. Using Northern blot analysis and a full-length PN1 cDNA probe we demonstrated the presence of PN1 mRNA in human term placentas. In situ hybridization of placental tissue with a PN1 riboprobe showed that PN1 mRNA is present throughout the placenta and is also abundant in the placental membranes. Immunohistochemical analysis with an anti-PN1 antibody showed co-localization of PN1 and its mRNA within the placenta.

Modulation of airway inflammation in cystic fibrosis. In vivo suppression of interleukin-8 levels on the respiratory epithelial surface by aerosolization of recombinant secretory leukoprotease inhibitor.

Based on the knowledge that neutrophil elastase (NE) in cystic fibrosis (CF) epithelial lining fluid (ELF) can induce human bronchial epithelial cells to express the gene for interleukin 8 (IL-8), an 8.5-kD neutrophil chemoattractant, we have evaluated CF ELF for the presence of IL-8, and investigated the ability of aerosolized recombinant secretory leukoprotease inhibitor (rSLPI) to suppress NE, and hence IL-8, levels on the respiratory epithelial surface in CF. Enzyme-linked immunoassay revealed 21.9 +/- 4.8 nM IL-8 in CF ELF compared with none in normals. Active NE was detectable in ELF of all individuals with CF and was significantly decreased (P < 0.03) after aerosolization of rSLPI. Human bronchial epithelial cells exposed to CF ELF recovered before rSLPI therapy expressed IL-8 mRNA transcripts, but ELF recovered after rSLPI therapy induced far less bronchial epithelial cell IL-8 gene expression. Consistent with this, rSLPI aerosol therapy caused a marked reduction in CF ELF IL-8 levels (P < 0.05) and neutrophil number (P < 0.02). There was also a clear association between CF ELF active NE and IL-8 levels (r = 0.94). These data suggest that rSLPI therapy not only suppresses respiratory epithelial NE levels, but also breaks a cycle of inflammation on the CF epithelial surface.

In vitro inhibitory effect of IL-8 and other chemoattractants on neutrophil-endothelial adhesive interactions.

We have previously reported that cytokine- or LPS-activated human umbilical vein endothelial cell (HUVEC) monolayers secrete IL-8 that can act as a neutrophil-selective adhesion inhibitor. In our study we investigated the mechanisms involved in the leukocyte adhesion inhibitory action of IL-8. The leukocyte adhesion inhibitory effect appears to be mediated by the action of IL-8 on the neutrophil, does not involve down-regulation of relevant endothelial adhesion molecules such as endothelial-leukocyte adhesion molecule-1 or intercellular adhesion molecule-1, and is quantitatively similar in different endothelial activation states that are predominantly endothelial-leukocyte adhesion molecule-1 dependent or intercellular adhesion molecule-1 dependent. In addition to inhibiting the attachment of freshly isolated peripheral blood neutrophils to cytokine-activated HUVEC monolayers, IL-8 also promoted a rapid detachment of tightly adherent neutrophils from activated HUVEC, and abolished neutrophil transendothelial migration. Certain other chemoattractants, including FMLP and C5a, had similar inhibitory actions, indicating IL-8 was not unique in its ability to inhibit various neutrophil-endothelial interactions. In contrast, two other neutrophil agonists 1-0-alkyl-2-acetyl sn-glycero-3-phosphocholine and granulocyte-macrophage-CSF, which, like IL-8, are produced by activated HUVEC, as well as the leukocyte-derived chemoattractant leukotriene B4, exerted minimal inhibitory effects on adhesion. Regardless of their ability to modulate neutrophil-endothelial cell adhesion, all these agents induced altered leukocyte surface expression of functionally important adhesion molecules, including loss of L-selectin (leukocyte adhesion molecule-1, LECAM-1) and increase in CD11b/CD18. Thus, although the above agonists have been characterized primarily as chemoattractants, our findings demonstrate that these agents can exert a wide range of modulatory effects on neutrophil-endothelial adhesive interactions.

Effect of acute inflammatory lung injury on the expression of monocyte chemoattractant protein-1 (MCP-1) in rat pulmonary alveolar macrophages.

Using a well-characterized rat model of immune complex-mediated acute inflammatory lung injury, we determined that there is a time-dependent elaboration of monocyte chemotactic activity in bronchoalveolar lavage fluid. Monocyte chemotactic activity is also significantly enhanced in culture supernatants from pulmonary alveolar macrophages (PAMs) from injured rat lungs. Northern hybridization analysis revealed markedly increased expression of rat monocyte chemoattractant protein 1 (MCP-1) mRNA in PAMs obtained from rats with immune complex-induced lung injury. The increased expression of MCP-1 mRNA and associated increase in monocyte chemotactic activity present in culture supernatants of PAMs from injured rat lungs suggest that PAMs may participate in the pathogenesis of acute inflammatory lung injury by the secretion of monocyte chemoattractants including MCP-1.

Structure-function analysis of hepatocyte growth factor: identification of variants that lack mitogenic activity yet retain high affinity receptor binding.

Hepatocyte growth factor (HGF) is a potent mitogen for parenchymal liver, epithelial and endothelial cells. Structurally, it has similarities to kringle-containing serine proteases, although it does not possess proteolytic activity. A structure-activity relationship study of human HGF was performed by functional analysis of HGF substitution and deletion variants. Analysis of HGF variants was accomplished by defining their ability to induce DNA synthesis on hepatocytes in primary culture and to compete with wild-type HGF for binding to a soluble form of the HGF receptor. Three groups of variants were made: (i) substitutions at the cleavage site, (ii) substitutions within the protease-like domain and (iii) deletions of the beta-chain and/or kringle domains. Our results show that: (i) single-chain HGF is a zymogen-like promitogen in that cleavage into a two-chain form is required for biological activity, however, the single chain form of HGF still retains substantial receptor binding capacity; (ii) certain mutations in the protease-like domain result in variants that are completely defective for mitogenic activity, yet exhibit apparent receptor binding affinities similar to wild-type HGF (Kd approximately 50-70 pM); and (iii) a variant containing the N-terminal 272 residues of mature HGF showed only a 4-fold increase in Kd when compared with wild-type HGF indicating that a primary receptor binding determinant is located within this sequence.

Effects of intravenous IL-8 administration in nonhuman primates.

IL-8, a cytokine known for its potent and specific neutrophil activation and chemoattractant properties, has been recently detected in the circulation during septic shock, endotoxemia, and after IL-1 alpha administration. Because of its observed in vitro actions, it has been hypothesized that IL-8 may contribute to the dynamics of circulating granulocytes and to the pathologic sequelae seen in sepsis. Here, human rIL-8 is administered to healthy nonhuman primates as a single i.v. injection or as a continuous 8-h i.v. infusion. We demonstrate that both methods of i.v. administration result in a rapid but transient, severe granulocytopenia, followed by a granulocytosis that persists as long as IL-8 levels are detectable in the circulation. There were no hemodynamic changes after IL-8 administration, and animals remained clinically stable during the 24-h observation period. No detectable circulating TNF-alpha, IL-1 beta, or IL-6 response was induced by either IL-8 administration regimen. Histopathologic examination revealed mild to moderate neutrophilic margination in lung, liver, and spleen, of greater severity in baboons receiving the 8-h infusion. There was no associated neutrophilic infiltration or tissue injury. Thus, IL-8 modulates circulating granulocyte dynamics and likely directs their actions, but when administered i.v. to healthy animals, either as a bolus dose or as a continuous infusion for up to 8 h, does not induce the hemodynamic and metabolic aberrations or the acute organ damage seen during sepsis.

Scanning mutagenesis of interleukin-8 identifies a cluster of residues required for receptor binding.

In order to identify residues required for the binding of interleukin-8 (IL-8) to its receptor, mutants were constructed in which clusters of charged amino acids were systematically replaced with alanine along the entire IL-8 sequence. The mutants were tested for their ability to induce a receptor-mediated rise in cytosolic free Ca2+, a property of wild-type IL-8 which can readily be detected by flow cytometry using neutrophils loaded with the calcium probe Indo-1. Eleven of the 12 mutants caused neutrophil calcium mobilization at 5 nM; the exception being a triple alanine mutant at positions K3, E4, and R6, which was inactive at all concentrations tested (150 nM maximum). A second set of mutants was generated in which residues 1-15 were individually mutated to alanine. Mutants E4A, L5A, or R6A were all inactive in the Ca2+ assay at 5 nM and competed poorly with 125I-IL-8 for neutrophil receptor binding; I10A, E4A, L5A, and R6A had approximately 30-, 100-, 100-, and 1000-fold reduced affinity, as compared with control IL-8, respectively. The nuclear magnetic resonance structure of IL-8 indicates that, in solution, the side chains of E4, L5, R6, and I10 point away from the core of the protein and do not participate in any intramolecular hydrogen bonds or salt bridges (Clore, G. M., and Gronenborn, A. M. (1991) J. Mol. Biol. 217, 611-620).

Red blood cells are a sink for interleukin 8, a leukocyte chemotaxin.

IL-8 (also known as neutrophil-activating peptide 1) is recognized as a potent effector of neutrophil functions. Several different cell types that contact blood, namely T lymphocytes, monocytes, and endothelial cells, secrete this polypeptide following stimulation by cytokines, or lipopolysaccharide. Here we show that when IL-8 is added to blood it rapidly partitions from the plasma fluid to the blood cells and that erythrocytes account for the vast majority of this binding. Analysis of 125I-IL-8 binding [( ala-IL-8]77 form) to human red cells indicates a single, 5 nM Kd affinity class of binding sites, present at approximately 2,000 per red cell representing approximately 15 nmol of red cell IL-8 binding sites per liter of blood. These sites are protease sensitive. Their binding of IL-8 is rapidly reversible and does not result in receptor internalization, although bound IL-8 is resistant to extraction by pH 3 buffer at 5 degrees C. 125I-IL-8 binding to red cells was not inhibited by epidermal growth factor or interleukin 1, but was inhibited by monocyte chemotactic peptide-1, which is not a neutrophil chemotaxin, but is a member of the same family of polypeptides as IL-8. FACS analysis of IL-8-mediated mobilization of Ca2+ in neutrophils indicates that the IL-8 bound to red cells is incapable of stimulating neutrophils. Thus, red cell absorption of IL-8 may function to limit stimulation of leukocytes by IL-8 released into blood.

Intravascular IL-8. Inhibitor of polymorphonuclear leukocyte accumulation at sites of acute inflammation.

IL-8 has been characterized primarily as a polymorphonuclear leukocyte (PMN) chemoattractant and proinflammatory mediator. Recently, we have reported that [Ala-IL-8]77 is secreted by activated cultured human endothelial cells and can function as a potent inhibitor of PMN adhesion to these monolayers. The pathophysiologic relevance of this in vitro observation was examined by determining the effects of intravascular or extravascular administration of IL-8 on PMN emigration at sites of acute inflammation in the skin of NZW rabbits. An i.v. bolus of [Ala-IL-8]77 (12 micrograms/kg) produced a marked and selective reduction of circulating PMN within 3 min, which returned toward preinjection levels within 30 min, and subsequently exceeded this level. A similar response was observed for circulating radiolabeled PMN, and gamma-scintigraphy determined that the lungs were the primary site of leukosequestration. During the 30- to 150-min interval after i.v. infusion of [Ala-IL-8]77, PMN emigration into acute inflammatory sites, elicited by various chemoattractants or cytokines, was significantly reduced, as judged histologically and quantitated with 51Cr-labeled PMN and myeloperoxidase measurements. Intravenous administration of [Ser-IL-8]72 yielded similar results. This inhibitory effect of i.v. IL-8 was transient and reinducible and did not reflect a suppression of the responsiveness of circulating PMN to chemoattractants. Intradermal injections of [Ala-IL-8]77 or [Ser-IL-8]72 induced dose-dependent PMN accumulation, which also was significantly reduced by i.v. administration of either form of IL-8. These results indicate that i.v. IL-8 can function as a PMN-directed leukocyte adhesion inhibitor and suggest that local secretion of IL-8 by activated endothelium may differentially modulate leukocyte-endothelial interactions at sites of acute inflammation.