Release Of Neutrophil Chemotactic Factors Research Articles

Cabozantinib Eradicates Advanced Murine Prostate Cancer by Activating Antitumor Innate Immunity.

Several kinase inhibitors that target aberrant signaling pathways in tumor cells have been deployed in cancer therapy. However, their impact on the tumor immune microenvironment remains poorly understood. The tyrosine kinase inhibitor cabozantinib showed striking responses in cancer clinical trial patients across several malignancies. Here, we show that cabozantinib rapidly eradicates invasive, poorly differentiated PTEN/p53-deficient murine prostate cancer. This was associated with enhanced release of neutrophil chemotactic factors from tumor cells, including CXCL12 and HMGB1, resulting in robust infiltration of neutrophils into the tumor. Critically, cabozantinib-induced tumor clearance in mice was abolished by antibody-mediated granulocyte depletion or HMGB1 neutralization or blockade of neutrophil chemotaxis with the CXCR4 inhibitor plerixafor. Collectively, these data demonstrate that cabozantinib triggers a neutrophil-mediated anticancer innate immune response, resulting in tumor clearance.Significance: This study is the first to demonstrate that a tyrosine kinase inhibitor can activate neutrophil-mediated antitumor innate immunity, resulting in invasive cancer clearance. Cancer Discov; 7(7); 750-65. ©2017 AACR.This article is highlighted in the In This Issue feature, p. 653.

Abstract 4979: Cabozantinib eradicates advanced murine prostate cancer by activating neutrophil-mediated anti-tumor innate immunity

Abstract Several kinase inhibitors targeting aberrant signaling pathways in tumor cells have been deployed in cancer therapy. However, their impact on the tumor immune microenvironment remains poorly understood. The tyrosine kinase inhibitor cabozantinib showed striking responses in cancer clinical trial patients across several malignancies. Here we show that cabozantinib rapidly eradicates invasive, poorly-differentiated PTEN/p53 deficient murine prostate cancer. This was associated with enhanced release of neutrophil chemotactic factors from tumor cells, including CXCL12 and HMGB1, resulting in robust infiltration of neutrophils into the tumor. Critically, cabozantinib-induced tumor clearance in mice was abolished by antibody-mediated granulocyte depletion or HMGB1 neutralization or blockade of neutrophil chemotaxis with the CXCR4 inhibitor, plerixafor. Collectively, these data demonstrate that cabozantinib triggers a neutrophil-mediated anti-tumor innate immune response, resulting in rapid tumor clearance. Citation Format: Akash Patnaik, Kenneth D. Swanson, Eva Csizmadia, Marina P. Gehring, Katja Helenius, Athalia R. Pyzer, Lily C. Wang, Jesse Novak, Olivier Elemento, Thomas B. Thornley, John M. Asara, John G. Clohessy, Kathy Kelly, Pier Paolo Pandolfi, Jacalyn M. Rosenblatt, David E. Avigan, Huihui Ye, Sabina Signoretti, Steven P. Balk, Lewis C. Cantley. Cabozantinib eradicates advanced murine prostate cancer by activating neutrophil-mediated anti-tumor innate immunity. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4979.

Surfactant Protein A Stimulates Release of Neutrophil Chemotactic Factors by Alveolar Type II Pneumocytes

Mucosal immunity is an important mechanism in the response to injury. Our hypothesis is that surfactant protein A (SP-A) is an autocrine factor that stimulates alveolar type II epithelial cell release of neutrophil chemotactic factors by binding to the SP-A receptor expressed by these cells. We examined (1) the effect of SP-A (20 μg/ml) or IL-1β (10 ng/ml) on release of neutrophil chemotactic factors by primary cultures of type II cells or alveolar macrophages, and (2) the effect of intratracheal instillation of the blocking antibody to the SP-A receptor on the response to oleic acid-induced lung injury in vivo. All media and cell culture supernates were assayed for neutrophil chemotactic activity, and bronchoalveolar lavage fluid from the in vivo experiments was analyzed for inflammatory cell counts. While SP-A and media used for the cell cultures has no intrinsic neutrophil chemotactic activity, supernates from primary cultures of type II cells incubated in either SP-A or IL-1β had twofold higher neutrophil chemotactic factor activity compared to supernates from controls. SP-A had no effect on release of neutrophil chemotactic factor by alveolar macrophages. Oleic acid-induced lung injury resulted in a marked influx of neutrophils into BAL, and this influx was reduced by 70% by pretreatment with the antibody to SP-A receptor. We conclude that SP-A stimulates the release of neutrophil chemotactic factor by alveolar type II cells, and this effect is mediated by the receptor for SP-A specifically expressed by these cells.

Methicillin-resistant Staphylococcus aureus: a new community-acquired pathogen?

The main goal of this review is to describe the emergence of methicillin-resistant Staphylococcus aureus (MRSA) as a community pathogen. Community-acquired MRSA has emerged as an important infection in the community setting. It has primarily been associated with skin and soft-tissue infections, but can also cause severe pulmonary infections, including pneumonia and empyema. Community-acquired MRSA is typically more susceptible to a wider class of antibiotics than healthcare-associated MRSA. Community-acquired MRSA is also more virulent compared with healthcare-associated MRSA isolates. Community-acquired MRSA usually contains the gene encoding Panton-Valentive leukocidin, which is a toxin that creates lytic pores in the cell membranes of neutrophils and induces the release of neutrophil chemotactic factors that promote inflammation and tissue destruction. The optimal antibiotic treatment for Panton-Valentive leukocidin-positive community-acquired MRSA is unknown; however, antibiotics with activity against MRSA and the ability to inhibit toxin production may be optimal (linezolid or clindamycin for susceptible isolates). Clinicians should be aware of the emergence of community-acquired MRSA as an important cause of serious infections arising in the community setting. Appropriate antibiotic therapy should be initiated as soon as infection with this pathogen is suspected.

Clostridium difficile toxin A induces the release of neutrophil chemotactic factors from rat peritoneal macrophages: role of interleukin-1beta, tumor necrosis factor alpha, and leukotrienes.

Clostridium difficile produces a potent enterotoxin and cytotoxin, toxins A and B, respectively, which appear to be responsible for pseudomenbranous colitis and antibiotic-associated diarrhea. In the present study we explored the neutrophil migration evoked by toxin A in the peritoneal cavities and subcutaneous air pouches of rats and examined the role of macrophages and their inflammatory mediators in this process. Toxin A causes a significant dose-dependent neutrophil influx into the peritoneal cavity, with a maximal response at 0.1 microg/ml and at 4 h. The depletion of macrophages by peritoneal washing prevents the toxin A-induced neutrophil migration into the peritoneal cavity. In contrast, an increase in macrophages induced by peritoneal injection of thioglycolate amplifies this toxin effect on neutrophil migration. Furthermore, the injection of supernatants from toxin A-stimulated macrophages into the rat peritoneal cavity causes significant neutrophil migration. Pretreatment of rats with BWA4C, nordihydroguaiaretic acid, mepacrine, or dexamethasone inhibits the neutrophil migration evoked by toxin A in the peritoneal cavities. However, pretreatment with the cyclooxygenase inhibitor indomethacin or the platelet-activating factor antagonist BN52021 fails to alter toxin A-induced neutrophil migration. Toxin A was also injected into air pouches of normal rats or rats pretreated with anti-interleukin-1beta (anti-IL-1beta) or anti-tumor necrosis factor alpha (anti-TNF-alpha) antibodies. Anti-TNF-alpha or anti-IL-1beta antibodies significantly reduce the neutrophil migration induced by toxin A. These data suggest that neutrophil migration evoked by toxin A is in part dependent on macrophage-derived cytokines, such as TNF-alpha and IL-1beta, and leukotrienes. These mediators may help to explain the intense inflammatory colitis caused by C. dificile toxin A in an experimental animal model of this disease.

Evidence of cardiac inflammation after open heart operations

In this study, 6 anesthetized dogs underwent global cardiac arrest for 1 hour, followed by reperfusion on bypass for 45 minutes. The hearts were then weaned off cardiopulmonary bypass and monitored for an additional 2 hours. Using modified Boyden chambers, high levels of neutrophil chemotactic activity were detected (using a checkerboard analysis) in the coronary sinus effluents obtained during cardiac arrest. The activity tended to decline during reperfusion. Assay of myeloperoxidase (a marker for neutrophils) revealed an accumulation of large numbers of neutrophils in the right (14 ± 1.1 × 10 4 cells/g wet weight) and left (16 ± 1 × 10 4 cells/g wet weight) ventricles after 2 hours of reperfusion. Light microscopy evaluation confirmed the presence of neutrophils, not only in the ventricles, but also in a greater number in the right and left atria. Electron microscopy study of these hearts revealed the presence of mild reversible changes, indicating good preservation of the hearts during arrest. Results of this study provide evidence for an acate inflammatory reaction that takes place after cardiac operations and suggest a role for myocardial tissues in the initiation of such a response through their release of neutrophil chemotactic factors.

Cornea-Derived Neutrophil Chemotactic Factors as Predictors of Corneal Inflammation

Acute corneal inflammation, induced by a variety of insults, is characterized by corneal opacity, infiltration of neutrophils, redness, pain, and corneal cell injury. Using isolated bovine corneas, we tested the effects of six compounds (i.e., citric acid, 2-butanone, 1-butanol, propyl paraben, glycerin, and talc) on corneal opacity, release of neutrophil chemotactic factors (NCFs) from corneal tissues, and release of lactic dehydrogenase (LDH) from corneal cells. The epithelial surfaces of isolated bovine corneas were incubated with the six compounds for 30 min and washed three times with saline, and the corneas were reincubated with culture medium for an additional 6 h at 37°C. Corneal opacity was measured quantitatively using an opacitometer machine. The levels of NCFs were measured in the supernatant using modified Boyden chambers. Although the effects of the six compounds on corneal opacity and release of NCF from corneal tissues were quite consistent with the in vivo data, the release of LDH was not...

Cardiac-derived neutrophil chemotactic factors: Detection in coronary sinus effluents of patients undergoing myocardial revascularization

Recent studies from our laboratory have demonstrated the release of neutrophil chemotactic factors from isolated rabbit hearts perfused with cardioplegic solutions and from ischemic dog hearts after coronary artery occlusion for 1 hour. On the basis of these animal studies, a test is now made of the hypothesis that neutrophil chemotactic factors are released by myocardial tissues of patients who undergo surgical myocardial revascularization. By means of modified Boyden chambers, the levels of neutrophil chemotactic factors were measured in effluent collected from the coronary sinuses of six patients undergoing cardiopulmonary bypass during periods of cold cardioplegia. Plain cardioplegic solutions were also analyzed. The standard formyl-methionyl-leucyl-phenylalanine, a stimulant of neutrophil recruitment, was used as a positive control solution. Results indicated the recovery of significantly high levels of neutrophil chemotactic factors in patient samples (i.e., 128% +/- 19% of formyl-methionyl-leucyl-phenylalanine) compared with control plain cardioplegic solution (less than 5% of formyl-methionyl-leucyl-phenylalanine) (p less than 0.0001). A standard checkerboard analysis indicated that the observed activity is chemotactic (i.e., directed migration) and not chemokinetic (i.e., random migration). This study also showed that these factors are proteins of a molecular weight in excess of 300 kd and exhibit in vivo activity by recruiting neutrophils into rabbit skin. The absence of immune cell-derived chemoattractants such as interleukin-1 and leukotriene B4 in these coronary sinus effluents suggests that the observed chemotactic activity is cardiac derived. Results of this investigation therefore demonstrate the release of neutrophil chemotactic factors by ischemic human hearts during cardiopulmonary bypass.

Cornea derived neutrophil chemotactic factors: intracellular synthesis and release

The present study was designed to test the hypothesis that the release of neutrophil chemotactic factors (NCF) from isolated corneas following hydrogen peroxide stimulation requires specific intracellular synthesis. For these studies, the epithelial surfaces of isolated rabbit corneas were preincubated with various inhibitors of protein synthesis (cycloheximide, 10 micrograms/ml, and puromycin, 50 micrograms/ml) and transcription (actinomycin D, 5 micrograms/ml) for 60 min prior to exposure of the corneas to glucose (G, 1 mg/ml) and glucose oxidase (GO, 20 U/ml) for 6 h at 37 degrees C. All three inhibitors decreased the levels of NCF recovered in the extracorneal fluids by 80-98%, suggesting that peroxide acts to upregulate NCF production at both the transcription and translation levels. When corneas were incubated with G/GO for 6 h at 12 degrees C or 4 degrees C instead of 37 degrees C, a reduction in the levels of NCF recovered in the supernatants was noted at 12 degrees C (46-91% inhibition) and at 4 degrees C (67-96% inhibition), suggesting that the synthesis of NCF at cold temperature was only reduced but not totally inhibited. To demonstrate that the observed reduction in chemotactic activity recovered from corneas incubated at 12 degrees C or 4 degrees C is not due to a temperature-dependent inhibition of NCF biosynthesis, but rather to a disruption of intracellular vesicular transport, temperature shift experiments were performed. Corneas were incubated with G/GO overnight at 12 degrees C or 4 degrees C prior to shifting to 37 degrees C for an additional 6 h.(ABSTRACT TRUNCATED AT 250 WORDS)