Neutrophilic leucocytosis induced by granulocyte colony-stimulating factor and interleukin-6 in canine primary lung adenocarcinoma.

BACKGROUND: Neutrophilic leucocytosis as a paraneoplastic syndrome may occur in dogs with lymphoma, renal carcinoma, rectal polyps and metastatic fibrosarcoma. However, the information on canine lung adenocarcinoma with neutrophilic leucocytosis is lacking. OBJECTIVE: This study aimed to describe the clinical features and cytokine profiles of canine patients with primary lung adenocarcinoma and neutrophilic leucocytosis. METHODS: Two dogs (cases #1 and #2), each with a solitary lung adenocarcinoma, were included. Both cases had leucocytosis and underwent lung lobectomy. The resected tumours were analysed for the expression of granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-6 (IL6) by quantitative real-time PCR compared with normal lung tissues. RESULTS: At the initial examination, neither patient had any clinical signs or fever. White blood cell count (WBC) was 58,300/µl and 32,900/µl in cases #1 and #2, respectively. The gene expression of G-CSF increased 6.7-and 19.7-fold in cases #1 and #2, respectively. The gene expression of IL6 markedly increased (30-fold) in case #1, whereas it increased slightly (1.9-fold) in case #2. On the other hand, that of GM-CSF was slightly changed in both cases. The WBC count postoperatively decreased to within the normal range in both cases. The postoperative survival times were 347 and 118 days in cases #1 and #2, respectively. CONCLUSIONS: This study describes G-CSF and IL6 producing lung adenocarcinoma associated with neutrophilic leucocytosis in dogs. Canine patients with pulmonary adenocarcinomas that have elevated G-CSF and IL6 levels may have a guarded prognosis. Further investigations are needed to clarify the prognosis of canine cytokine-producing lung adenocarcinoma.

Commensal Microbiome Expands Tγδ17 Cells in the Lung and Promotes Particulate Matter-Induced Acute Neutrophilia.

Particulate matter (PM) induces neutrophilic inflammation and deteriorates the prognosis of diseases such as cardiovascular diseases, cancers, and infections, including COVID-19. Here, we addressed the role of γδ T cells and intestinal microbiome in PM-induced acute neutrophilia. γδ T cells are a heterogeneous population composed of Tγδ1, Tγδ2, Tγδ17, and naïve γδ T cells (TγδN) and commensal bacteria promote local expansion of Tγδ17 cells, particularly in the lung and gut without affecting their Vγ repertoire. Tγδ17 cells are more tissue resident than Tγδ1 cells, while TγδN cells are circulating cells. IL-1R expression in Tγδ17 cells is highest in the lung and they outnumber all the other type 17 cells such as Th17, ILC3, NKT17, and MAIT17 cells. Upon PM exposure, IL-1ß-secreting neutrophils and IL-17-producing Tγδ17 cells attract each other around the airways. Accordingly, PM-induced neutrophilia was significantly relieved in γδ T- or IL-17-deficient and germ-free mice. Collectively, these findings show that the commensal microbiome promotes PM-induced neutrophilia in the lung via Tγδ17 cells.

Pharmacological targeting of host chaperones protects from pertussis toxin in vitro and in vivo.

Whooping cough is caused by Bordetella pertussis that releases pertussis toxin (PT) which comprises enzyme A-subunit PTS1 and binding/transport B-subunit. After receptor-mediated endocytosis, PT reaches the endoplasmic reticulum from where unfolded PTS1 is transported to the cytosol. PTS1 ADP-ribosylates G-protein α-subunits resulting in increased cAMP signaling. Here, a role of target cell chaperones Hsp90, Hsp70, cyclophilins and FK506-binding proteins for cytosolic PTS1-uptake is demonstrated. PTS1 specifically and directly interacts with chaperones in vitro and in cells. Specific pharmacological chaperone inhibition protects CHO-K1, human primary airway basal cells and a fully differentiated airway epithelium from PT-intoxication by reducing intracellular PTS1-amounts without affecting cell binding or enzyme activity. PT is internalized by human airway epithelium secretory but not ciliated cells and leads to increase of apical surface liquid. Cyclophilin-inhibitors reduced leukocytosis in infant mouse model of pertussis, indicating their promising potential for developing novel therapeutic strategies against whooping cough.

Bivalent Inhibitor with Selectivity for Trimeric MMP-9 Amplifies Neutrophil Chemotaxis and Enables Functional Studies on MMP-9 Proteoforms.

A fundamental part of the immune response to infection or injury is leukocyte migration. Matrix metalloproteinases (MMPs) are a class of secreted or cell-bound endopeptidases, implicated in every step of the process of inflammatory cell migration. Hence, specific inhibition of MMPs is an interesting approach to control inflammation. We evaluated the potential of a bivalent carboxylate inhibitor to selectively inhibit the trimeric proteoform of MMP-9 and compared this with a corresponding monovalent inhibitor. The bivalent inhibitor efficiently inhibited trimeric MMP-9 (IC50 = 0.1 nM), with at least 500-fold selectivity for MMP-9 trimers over monomers. Surprisingly, in a mouse model for chemotaxis, the bivalent inhibitor amplified leukocyte influxes towards lipopolysaccharide-induced inflammation. We verified by microscopic and flow cytometry analysis increased amounts of neutrophils. In a mouse model for endotoxin shock, mice treated with the bivalent inhibitor had significantly increased levels of MMP-9 in plasma and lungs, indicative for increased inflammation. In conclusion, we propose a new role for MMP-9 trimers in tempering excessive neutrophil migration. In addition, we have identified a small molecule inhibitor with a high selectivity for the trimeric proteoform of MMP-9, which will allow further research on the functions of MMP-9 proteoforms.

A Combination of Kidney Ischemia and Injection of Isolated Mitochondria Leads to Activation of Inflammation and Increase in Mortality Rate in Rats.

We studied the development of acute kidney injury and animal death in the model of combined injury caused by kidney ischemia/reperfusion with simultaneous systemic administration of mitochondria. It was found that intraperitoneal injection of isolated mitochondria led to the appearance of mitochondrial DNA in the peripheral blood that could activate innate immunity. After administration of mitochondria, as well as after renal ischemia/reperfusion, proinflammatory changes were observed, primarily leukocytosis and granulocytosis. The combination of ischemia/reperfusion with injection of mitochondria caused a sharp increase in animal death, which may indicate a direct link between activation of TLR-signaling and high mortality of patients with combined injuries and multiple-organ failure in intensive care units. Treatment with mitochondria-targeted antioxidant increased animal survival, which indicated the participation of mitochondrial ROS in the development of systemic inflammatory response and death caused by acute renal failure.

Preoperative Leukocytosis as a Prognostic Marker in Endometrioid-Type Endometrial Cancer: A Single-Center Experience from Saudi Arabia.

INTRODUCTION: Only a few studies (n=5) have focused on the importance of preoperative high white blood cell (WBC) count (leukocytosis) as a prognostic marker in patients with endometrial cancer (EC). Nevertheless, more related studies are needed to solidly corroborate these findings. To the best of our knowledge, no such study has been conducted in the Gulf region and Saudi Arabia in particular. METHODS: A retrospective cross-sectional study was conducted at King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia. The medical records of 130 patients with endometrioid-type EC were reviewed for clinico-pathological factors (that is, age, tumor stage, endometrioid grade, myometrial invasion depth, lymphovascular space involvement and recurrence) and survival outcomes. Survival outcomes included disease-free survival (DFS) and overall survival (OS). Leukocytosis was defined as a WBC count level >10 x 103 cells/uL. Chisquare test was used for univariate analysis of categorical data. Survival analyses of DFS and OS were calculated according to the Kaplan-Meier estimates method and compared by using two-tailed log-rank test. Univariate and multivariate analyses of survival were performed using Cox proportional hazards model. Statistical significance was regarded as a p value <0.05. RESULTS: The mean age was 59 ± 10.5 years (range: 36-99). The overall mean preoperative WBC count was 7.7 ± 2.4 x 103 cells/uL (range: 2.7-17 x 103). The frequency of preoperative leukocytosis was 18.5% (n=24). Patients with preoperative leukocytosis have statistically significant higher rates of advanced FIGO stage III-IV disease (p=0.007) and positive tumor recurrence (p=0.009) than patients with normal preoperative WBC count (chisquare test). Patients with preoperative leukocytosis have a higher statistically significant probability of developing recurrence than patients with preoperative normal WBC count (29.4 vs. 11.8%, p=0.008, log-rank test). Patients with preoperative leukocytosis have statistically significant lower mean DFS (58.3 ± 6.9 vs. 67.9 ± 2.3 months, p=0.015) and 5-year DFS rate (66.7 vs. 86.8%, p=0.015) than patients with normal preoperative WBC counts (log-rank test). However, there were no statistically significant differences between patients with preoperative leukocytosis and normal WBC counts in terms of mean OS (73.8 ± 4.5 vs. 79.3 ± 2.1, p=0.581) and 5-year OS rate (87.5 vs. 91.5%, p=0.581), respectively (log-rank test). Multivariate analyses using Cox proportional hazards model failed to significantly demonstrate preoperative WBC count as an independent prognostic factor of DFS and OS (log-rank test, p>0.05). CONCLUSION: Preoperative leukocytosis is not rare in patients with endometrioid-type EC. Besides, preoperative leukocytosis is correlated with poor tumor FIGO stage, higher cumulative incidence of relapse and poor DFS in the univariate analysis. Our study suggests that preoperative leukocytosis may identify high-risk patients who may require more intensified therapy in terms of aggressive debulking and/or perioperative chemotherapy.

Tumor immune microenvironment in cancer patients with leukocytosis.

Tumor-related leukocytosis (TRL) is correlated with poor survival in various types of cancers, but the microenvironment of TRL-associated human tumors has not been fully elucidated. Here, we aimed to characterize the immune microenvironment of cancer patients with TRL. The transcriptional signatures of tumor tissues obtained from cervical cancer patients with (TRLpos) and without TRL (TRLneg) were compared. As a surrogate for TRL diagnosis, a leukocytosis signature (LS) score was derived using genes differentially expressed between TRLpos and TRLneg tumors. The immunological profiles of patients in the TCGA database with high (LShigh) or low LS scores were compared. TRLpos tumors were transcriptionally distinct from TRLneg tumors, exhibiting up-regulation of radioresistance and down-regulation of adaptive immune response-related genes. In the TCGA cervical cancer cohort (n = 303), patients with high LS had inferior survival rates compared to those with low LS (P = 0.023). LShigh tumors were enriched in radioresistance, wound healing, and myeloid-derived suppressor cell (MDSC) signatures and had a higher infiltration of M2 macrophages and a lower infiltration of M1 macrophages and lymphocytes. LShigh tumors also expressed higher levels of CXCR2 chemokines, CSF2, and CSF3. In the pan-cancer cohort (n = 9984), LShigh tumors also exhibited poor survival, signatures of a suppressive immune microenvironment, and higher expression of CXCR2 chemokines. Our data provide evidence for a suppressive immune microenvironment in patients with TRL and suggest promising targets, such as the CXCR2 axis, for its therapeutic intervention.

GATA2 hypomorphism induces chronic myelomonocytic leukemia in mice.

The transcription factor GATA2 regulates normal hematopoiesis, particularly in- stem cell maintenance and myeloid differentiation. Various heteroallelic GATA2 gene mutations are associated with a variety of hematological neoplasms, including myelodysplastic syndromes and leukemias. Here, we report that impaired GATA2 expression induces myelodysplastic and myeloproliferative neoplasm development in elderly animals, and this neoplasm resembles chronic myelomonocytic leukemia in humans. GATA2 hypomorphic mutant (G2f GN / fGN ) mice that were generated by the germline insertion of a neocassette into the Gata2 gene locus avoided the early embryonic lethality observed in Gata2-null mice. However, adult G2f GN / fGN mice suffered from exacerbated leukocytosis concomitant with progressive anemia and thrombocytopenia and eventually developed massive granulomonocytosis accompanied by trilineage dysplasia. The reconstitution activity of G2f GN / fGN mouse stem cells was impaired. Furthermore, G2f GN / fGN progenitors showed myeloid lineage-biased proliferation and differentiation. Myeloid progenitor accumulation started at a younger age in G2f GN / fGN mice and appeared to worsen with age. G2f GN / fGN mice showed increased expression of transcripts encoding cytokine receptors, such as macrophage colony-stimulating factor receptor and interleukin-6 receptor, in granulocyte-monocyte progenitors. This increased expression could be correlated with the hypersensitive granulomonocytic proliferation reaction when the mice were exposed to lipopolysaccharide. Taken together, these observations indicate that GATA2 hypomorphism leads to a hyperreactive defense response to infections, and this reaction is attributed to a unique intrinsic cell defect in the regulation of myeloid expansion that increases the risk of hematological neoplasm transformation.

TNF, but not hyperinsulinemia or hyperglycemia, is a key driver of obesity-induced monocytosis revealing that inflammatory monocytes correlate with insulin in obese male mice.

Inflammation contributes to obesity-related hyperinsulinemia and insulin resistance, which often precede type 2 diabetes. Inflammation is one way that obesity can promote insulin resistance. It is not clear if the extent of obesity, hyperinsulinemia, or hyperglycemia, underpins changes in cellular immunity during diet-induced obesity. In particular, the requirement for obesity or directionality in the relationship between insulin resistance and monocyte characteristics is poorly defined. Inflammatory cytokines such as tumor necrosis factor (TNF) can contribute to insulin resistance. It is unclear if TNF alters monocytosis or specific markers of cellular immunity in the context of obesity. We measured bone marrow and blood monocyte characteristics in WT and TNF-/- mice that were fed obesogenic, high fat (HF) diets. We also used hyperglycemic Akita mice and mice implanted with insulin pellets in order to determine if glucose or insulin were sufficient to alter monocyte characteristics. We found that diet-induced obesity in male mice increased the total number of monocytes in blood, but not in bone marrow. Immature, inflammatory (Ly6Chigh ) monocytes decreased within the bone marrow and increased within peripheral blood of HF-fed mice. We found that neither hyperinsulinemia nor hyperglycemia was sufficient to induce the observed changes in circulating monocytes in the absence of diet-induced obesity. In obese HF-fed mice, antibiotic treatment lowered insulin and insulin resistance, but did not alter circulating monocyte characteristics. Fewer Ly6Chigh monocytes were present within the blood of HF-fed TNF-/- mice in comparison to HF-fed wild-type (WT) mice. The prevalence of immature Ly6Chigh monocytes in the blood correlated with serum insulin and insulin resistance irrespective of the magnitude of adipocyte or adipose tissue hypertrophy in obese mice. These data suggest that diet-induced obesity instigates a TNF-dependent increase in circulating inflammatory monocytes, which predicts increased blood insulin and insulin resistance independently from markers of adiposity or adipose tissue expansion.

The management of hyperleukocytosis in 2017: Do we still need leukapheresis?

Hyperleukocytosis is defined as a white blood cell count greater than 100.000/µL in patients affected by acute or chronic leukemias. Hyperleukocytosis is more common in acute leukemias than in chronic leukemias. Risk factors include younger age, acute myeloid leukemia, the microgranular variant of acute promyelocytic leukemia, acute lymphoblastic leukemia and some cytogenetic abnormalities. Although it can affect any organ system, symptoms usually arise from involvement of the cerebral, pulmonary and renal microvasculature. The term "leukostasis" refers to 'symptomatic hyperleukocytosis' which is a medical emergency that needs prompt recognition and initiation of therapy to prevent renal and respiratory failure or intracranial haemorrhage. The underlying mechanisms of hyperleukocytosis and leukostasis are poorly understood. The management of hyperleukocytosis and leukostasis involves supportive measures and reducing the number of circulating leukemic blast cells by induction chemotherapy, hydroxyurea, low-dose chemotherapy, and leukapheresis. The measures such as hydroxyurea, low-dose chemotherapy, and leukapheresis shouldn't be considered to correct the laboratory abnormalities in patients with hyperleukocytosis who have no signs or symptoms. Also, neither hydroxyurea nore leukapheresis is able to show benefit on short and long term outcomes in patients with symptomatic hyperleukocytosis. The optimal management of symptomatic hyperleukocytosis is still uncertain, and there are no randomized studies demonstrating one is superior to each other. Therefore, it is recommended that intensive chemotherapy should be implemented as quickly as possible in treatment-eligible patients, in parallel with supportive measures for DIC and TLS.

Changes in platelet parameters in leukocytosis.

INTRODUCTION: In recent years, platelets are known to have a large variety of functions in many pathophysiological processes and their interaction with endothelial cells and leukocytes is known to play an important role in the pathophysiology of vascular inflammation. The aim of this study was to investigate the relationship between white blood cell count in conditions resulting in leukocytosis and platelet count and platelet parameters including mean platelet volume, platelet distribution width, and plateletcrit. METHODS: White blood cell counts count and all platelet parameters were evaluated in 341 results of normal complete blood count (of which the white blood cell counts were within reference range, group 1) and 327 results of elevated white blood cell counts count (group 2). RESULTS: There was a significant difference between these two groups in PLT counts and PCT values, being higher in Group 2. However, there was no statistically significant difference between two groups in MPV and PDW values. On the other hand, there were statistically significant, but weak, correlations between the WBC and platelet counts in both groups (p<0.01, r=0.235 for group 1, p<0.05, r=0.116 for group 2). CONCLUSION: As a conclusion PLT count and PCT values increase in infectious conditions. This study and previous studies show that PLTs are employed in infectious conditions but the exact mechanism and the exact clinical importance of this response remains to be cleared by further studies.

[Oxygen Leukocyte Larceny]. / Sequestro Leucocitário de Oxigénio.

The authors present a case of a 60-year-old male patient, previously diagnosed with B-cell chronic lymphocytic leukemia, who was admitted to the Emergency Room with dyspnea. The initial evaluation revealed severe anemia (Hgb = 5.0 g/dL) with hyperleukocytosis (800.000/µL), nearly all of the cells being mature lymphocytes, a normal chest X-ray and a low arterial oxygen saturation (89%; pulse oximetry). After red blood cell transfusion, Hgb values rose (9.0 g/dL) and there was a complete reversion of the dyspnea. Yet, subsequent arterial blood gas analysis, without the administration of supplemental oxygen, systematically revealed very low oxygen saturation values (~ 46%), which was inconsistent with the patientâs clinical state and his pulse oximetry values (~ 87%), and these values were not corrected by the administration of oxygen via non-rebreather mask. The investigation performed allowed to establish the diagnosis of oxygen leukocyte larceny, a phenomenon which conceals the true oxygen saturation due to peripheral consumption by leukocytes.

Os autores apresentam um caso clínico de um homem de 60 anos, com diagnóstico de leucemia linfocítica crónica-B, observado no Serviço de Urgência por dispneia de esforço. Na avaliação inicial, destacava-se uma anemia grave (Hgb = 5,0 g/dL) e hiperleucocitose (800 000/µL), correspondendo na sua quase totalidade a linfócitos maduros, uma radiografia do tórax normal e uma saturação de oxigénio baixa (89%). Após transfusão de glóbulos rubros registou-se uma elevação do valor de Hgb (9,0 g/dL) e uma reversão completa do quadro de dispneia. Porém, as gasimetrias subsequentes de reavaliação, em ar ambiente, revelaram sistematicamente valores de saturação de oxigénio muito baixos (~ 46%), discrepantes com o estado clínico do doente e com a oximetria de pulso (~ 87%), e não sofreram qualquer alteração com administração de concentrações de oxigénio por máscara. A investigação efetuada permitiu estabelecer o diagnóstico de sequestro leucocitário de oxigénio, fenómeno que mascara a verdadeira saturação de oxigénio por consumo periférico pelos leucócitos.

Age-associated vascular inflammation promotes monocytosis during atherogenesis.

Aging leads to a proinflammatory state within the vasculature without disease, yet whether this inflammatory state occurs during atherogenesis remains unclear. Here, we examined how aging impacts atherosclerosis using Ldlr(-/-) mice, an established murine model of atherosclerosis. We found that aged atherosclerotic Ldlr(-/-) mice exhibited enhanced atherogenesis within the aorta. Aging also led to increased LDL levels, elevated blood pressure on a low-fat diet, and insulin resistance after a high-fat diet (HFD). On a HFD, aging increased a monocytosis in the peripheral blood and enhanced macrophage accumulation within the aorta. When we conducted bone marrow transplant experiments, we found that stromal factors contributed to age-enhanced atherosclerosis. To delineate these stromal factors, we determined that the vasculature exhibited an age-enhanced inflammatory response consisting of elevated production of CCL-2, osteopontin, and IL-6 during atherogenesis. In addition, in vitro cultures showed that aging enhanced the production of osteopontin by vascular smooth muscle cells. Functionally, aged atherosclerotic aortas displayed higher monocyte chemotaxis than young aortas. Hence, our study has revealed that aging induces metabolic dysfunction and enhances vascular inflammation to promote a peripheral monocytosis and macrophage accumulation within the atherosclerotic aorta.

Absceso hepático secundario a una perforación duodenal por espina de pescado: descripción de un caso / Liver abscess secondary to duodenal perforation by fishbone: report of a case

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