Higher Percentage Of Lymphocytes Research Articles

Immunophenotypic characterization of double positive T lymphocytes in Cuban older adults.

Immunosenescence is the functional deterioration of the immune system during natural aging that mainly affects T lymphocytes, within which is the subpopulation of peripheral double positive T lymphocytes that are immunomodulators of the immune response. They are increased in older adults and play a fundamental role in the immunopathogenesis of autoimmune diseases, viral infections and neoplasms. To characterize the immunophenotype of peripheral double positive T lymphocytes in Cuban older adults. A cross-sectional study was carried out in 30 older adults residing in Cuba. Peripheral blood double positive T lymphocytes were quantified by flow cytometry. The reading was done on a Beckman Coulter Gallios flow cytometer. A Beckman Coulter Gallios flow cytometer was used for data analysis. Percentage values, median and standard deviation were used. There was a predominance of women who represented 70%. No older adults with low values of double positive T lymphocytes were reported. Women with high percentage and absolute values of DPT lymphocytes predominated in relation to men. Among the high percentage values and the absolute value of DPT lymphocytes, the group ≥80years prevailed. Most of the older adults who exhibited high percentage and absolute values of DPT lymphocytes had some associated comorbidity, an idea that suggests that CD4+CD8+ T cells seem to play an important role in peripheral sites as powerful immunosuppressants or as cells with high cytotoxic potential.

Prognostic significance of bronchoalveolar lavage cellular analysis in patients with acute exacerbation of interstitial lung disease

BackgroundAcute exacerbation (AE) of interstitial lung disease (ILD) is an acute respiratory deterioration of unknown etiology, associated with high mortality. Currently, bronchoalveolar lavage (BAL) has been no longer required for the diagnosis of AE-ILD; however, the clinical utility of BAL fluid (BALF) cellular analysis in AE-ILD remains unclear. MethodsA retrospective study of 71 patients who underwent BAL at our institution between 2005 and 2019 and were diagnosed with AE-ILD was conducted. We performed BALF cellular analysis and evaluated its prognostic significance. ResultsThere were 26 patients with AE of idiopathic pulmonary fibrosis (IPF) and 45 with AE of non-IPF, including idiopathic interstitial pneumonias/non-IPF (n = 22), ILD associated with collagen tissue disease (n = 20) and fibrotic hypersensitivity pneumonia (n = 3). All patients were treated with high-dose corticosteroids, and the 90-day mortality after AE was 31%. Most patients showed a high percentage of lymphocytes and/or neutrophils in BALF regardless of the underlying ILD. There was a significant negative correlation between BALF neutrophils and the PaO2/FiO2 ratio, and patients with UIP pattern or diffuse AE pattern on HRCT had a significantly higher percentage of BALF neutrophils than those with other patterns. Multivariate analysis revealed that lower and higher percentage of lymphocytes and neutrophils, respectively, in BALF were independent poor prognostic factors for 90-day survival. BALF lymphocyte and neutrophil count ≥25% and <20%, respectively, predicted favorable survival after AE. ConclusionsCellular analysis of BALF in AE-ILD is a potential biomarker for predicting prognosis after AE.

Prognostic Value of Absolute Lymphocyte count, Lymphocyte percentage, Serum albumin, Aberrant Expression of CD7, CD19 and the Tumor Suppressors (PTEN and p53) in Patients with Acute Myeloid Leukemia

Background: Acute myeloid leukemia (AML) is a hematopoietic neoplasm. Tumor suppressors have a magnificent role in preventing the AML process. The absolute lymphocyte count is a simple yet statistically powerful estimate in patients with acute leukemia besides the lymphocyte’s percentage. Aim: Investigating the prognostic value of absolute lymphocyte count, lymphocyte percentage, serum albumin, the aberrant expression of CD7and CD19 and the tumor suppressor genes (PTEN and p53) in patients with AML. Methods: 35 de novo AML patients were included. They received the standard induction chemotherapy (3+7 protocol) and were followed up for one year after treatment. 15 normal healthy individuals, age and sex matched constituted the controls.Results: The mean overall survival of patients with lymphocyte percentage ≤25 was low compared to those with high lymphocyte percentage (&gt;25%) (χ2 =5.808, P=0.016). AML patients with low levels of ALC showed significantly shorter overall survival than patients with high levels (χ2 =4.587, P= 0.032). AML patients with low serum albumin were of low overall survival compared to those with normal level (χ2 =8.698, P=0.003). Patients with aberrant CD7 expression showed short survival and unresponsiveness to treatment than CD7 negative patients. PTEN gene expression and p53 protein level were significantly lower in AML patients compared to the control group.Conclusion: The decrease in ALC, lymphocyte percentage, albumin concentration and the increase in monocyte percentage indicates bad prognosis in AML patients. The Aberrant CD7 expression, very low expression of PTEN and low level of p53 could estimate the unresponsiveness to standard chemotherapy.

IMMU-21. INVESTIGATION OF WHITE BLOOD CELL CHARACTERISTICS IN CSF SAMPLES AT PEDIATRIC BRAIN TUMOR DIAGNOSIS

BACKGROUNDThere has been a recent surge in investigation of immunity and immunotherapy, but their role in pediatric brain tumors is incompletely defined. We hypothesized that investigating an understudied dataset, WBC and differential results in CSF drawn at the time of pediatric brain tumor diagnosis to look for microscopic metastases, would provide insight into the role of immunology and potential for immunotherapy in these diseases and correlate with prognosis and/or metastasis.METHODSWe conducted a retrospective comparison analysis of CSF values in 349 patients at our institution from samples drawn within 60 days of initial CNS tumor diagnosis from 1998–2018. We examined total nucleated cell count, absolute counts and percentages for WBC subtypes. We compared CSF values by tumor cell presence, patient vital status, and disease group: atypical teratoid rhabdoid tumor, ependymoma, germinoma, high-grade glioma (HGG), low-grade glioma (LGG), medulloblastoma, non-germinomatous germ cell tumor, and other embryonal tumors (OET). We used Wilcoxon and Kruskal-Wallis tests for comparisons.RESULTSOverall, higher lymphocyte percentage (p=0.002) and lower monocyte percentage (p=0.007) were associated with survival. WBC characteristics did not differ significantly based on tumor cell presence. Compared to medulloblastoma, ependymoma showed a more active CSF immune response, while LGG, HGG, and OET showed a less active response, based on total WBC and/or absolute neutrophil count (p=0.001–0.007).CONCLUSIONSHigher lymphocyte and lower monocyte percentages in CSF correlated with better prognosis overall; causality requires further investigation. Tumor subtypes varied in their immune stimulation, offering potential insight into which will be amenable to immunotherapy.

Profile of Checkpoint Molecules Expression on Bone Marrow Cell Populations in Patients with High-Risk Myelodysplastic Syndrome

Background Immune checkpoint (IC) inhibitors (ICI) is a promising group of agents with potential effect in myelodysplastic syndrome (MDS). However, there is not enough data on the pattern of IC expression in MDS bone marrow, though this can guide future therapies with ICI. Methods We prospectively included 57 patients with high risk MDS during 2019-2020 years after signing informed consent for participation in research. We used 7 eight-colored panels in each patient to define IC expression on T- and NK-cells (CD3, CD8, CD4, CD56, CD16), myeloid precursors (CD117, CD34), T-regulators (CD25, CD4), myeloid-derived suppressor cells (CD15, CD11b, CD14, HLA-DR, CD33). In each population we assessed expression of CD279, CD152, CD223, TIM3, CD273, CD274, CD275, CD80. Results In lymphoid populations we observed an extremely high percentage of PD-1 positive lymphocytes (10.5 ± 8.0% of all nucleated cells), including PD-1 positive CD4 lymphocytes (6.3 ± 3.7%) and CD8 lymphocytes (4.7 ± 3.5%). Interestingly, on average 63 ± 34% of all lymphocytes were PD-1 positive (Figure 1A), which indicates an extreme degree of T-cell exhaustion and significant exploitation of the PD-1 system by a malignant tumor. Among the PD-1 ligands PD-1L predominated, however it was present only on a small percentage of myeloid precursors (2.2 ± 0.39% of nucleated cells), but most of the PD-1L ligand expression was observed on granulocytes in the process of their differentiation from myeloid precursors (5.8 ± 4.1% of nucleated cells), and in some patients the proportion of such granulocytes reached more than 20%. Moreover, on the cells of the monocytic series, PD-1L is practically not expressed, but CD80 is expressed, a ligand for CTLA4 (1.6 ± 0.7% of nucleated cells). Interestingly, despite the presence of expression of CD80 on monocytic and dendritic cells, the receptor for this ligand on T-lymphocytes was practically absent. The proportion of such cells was only 0.02 ± 0.01%, i.e. the expression of CD80 facilitates the activation of lymphocytes through interaction with CD28 rather than the suppression of the immune response through CD152. Thus, it can be suggested that CTLA4 does not have a clinically significant role in MDS. At the same time, a certain number of TIM3 positive lymphocytes were observed, the proportion of which was 0.42 ± 0.16%, and the percentage of lymphocytes carrying this receptor was 3.6 ± 1.2% (Figure 1A). However, significantly greater TIM3 expression was detected on NK cells. The proportion of such cells was 1.3 ± 0.6% of all nucleated cells, while 30% of all NK cells carried the TIM3 receptor (Figure 1A). Only 0.1% of cells from early myeloid progenitors that expressed TIM3 receptor were detected during flow cytometry, while the proportion of TIM3 positive myeloid cells increased during maturation just in the same way as the PD-1L. Expression of Gal9 was negligible, as it is a protein secreted into the intercellular space. Only a small percentage is associated with cell membranes. The analysis of the correlation matrix showed synchronous changes in the number of most bone marrow T-cell populations. A negative correlation of the level of myeloid tumor precursors actively expressing IC ligands and activated T cells, expressing CD223, NK cells, NKT was observed. The total number of myeloid tumor precursors positively correlated with the number of PD-1 positive lymphocytes (Figure 1B). Conclusion It was found that IC system plays a role in high-risk MDS. Moreover, the expression of ligands is mainly realized during maturation of blast cells into granulocytes, which are the main population expressing these inhibitory molecules. The leading role in the suppression of the T-cell response in MDS is played by the PD-1-PD-1L system, the CTLA4 system plays a minor role in our study cohort. The second most important system is GAL9-TIM3, effecting predominantly NK cells, while T cell are less affected. Thus, the data obtained justify the trials of dual blockade with anti-PD-1 and anti-TIM3 agents in high-risk MDS. Figure 1 Disclosures No relevant conflicts of interest to declare.

Clinical significance of serum anti-granulocyte\u2013macrophage colony-stimulating factor autoantibodies in patients with sarcoidosis and hypersensitivity pneumonitis

BackgroundAnti-granulocyte–macrophage colony-stimulating factor autoantibody (GMAb) has been recognized as a diagnostic biomarker for autoimmune pulmonary alveolar proteinosis (aPAP). The aims of this study were to know the incidence of increased level of serum GMAb in granulomatous lung diseases (sarcoidosis and hypersensitivity pneumonitis [HP]) and to clarify the role of GMAb. Consecutive individuals diagnosed with sarcoidosis (n = 92) and HP (n = 45) at National Hospital Organization Kinki-Chuo Chest Medical Center were retrospectively analyzed. We measured serum GMAb levels at the diagnosis. Cut-off values of GMAb discriminating aPAP (n = 110) from healthy controls (n = 31) were determined by receiver operating characteristic (ROC) curve analysis. We compared the clinical features of sarcoidosis and HP patients with GMAb levels above the cut-off value (“Elevated-GMAb”) with those of patients whose GMAb levels below the cut-off value (“Low-GMAb”). Radiological and pathological findings in elevated-GMAb patients were re-evaluated to elucidate the role of GMAb in granulomatous lung diseases.ResultsAnalysis of ROC indicated a sensitivity and specificity of 100% at GMAb level of 3.33 μg/mL for discriminating aPAP from healthy controls (area under curve = 1.000, p < 0.0001). The percentages of elevated-GMAb sarcoidosis and HP patients were 5.4% (n = 5) and 11.1% (n = 5), respectively. The number of comorbid sarcoidosis and HP patients with aPAP was two and one, respectively. Elevated-GMAb sarcoidosis patients presented with significantly higher serum levels of Krebs von den Lungen (KL)-6, surfactant protein-D (SP-D), lactate dehydrogenase, and the requirement of systemic corticosteroid therapy. Elevated-GMAb HP patients demonstrated older age, higher serum KL-6, SP-D, carcinoembryonic antigen, and cytokeratin fragment 21-1 levels, and a higher percentage of lymphocytes in bronchoalveolar lavage than low-GMAb patients. A subset of patients presented with radiological and pathological findings characteristic of aPAP.ConclusionsWe demonstrated the percentage of elevated-GMAb sarcoidosis and HP patients who presented with several features suggestive of aPAP. Elevated-GMAb sarcoidosis and HP patients without definitive aPAP diagnosis may have subclinical or early-stage aPAP and may not necessarily indicate false positives. Upon diagnosis of sarcoidosis or HP, measurement of GMAb may be useful in detecting possible comorbidity of subclinical or early-onset aPAP.

Efficacy and survival of infliximab in psoriasis patients: A single-center experience in China.

Psoriasis is a chronic, systemic disease that requires long-term management. Biologic agents have been used widely against psoriasis, such as infliximab. We analyzed the survival and discontinuation rates of infliximab when treating psoriasis under real-world conditions in China. Patients with moderate-to-severe psoriasis treated with infliximab in Shanghai Skin Disease Hospital from January 2015 to April 2020 were included in our retrospective study. Information from their medical records (clinical characteristics, Psoriasis Area Severity Index [PASI] score, laboratory results, and time of discontinuation) was collected through the Shanghai Skin Disease Hospital database. The survival of infliximab was assessed with Kaplan-Meier plots and multivariate Cox regression. Forty-two patients who underwent treatment were assessed retrospectively (38.1% had been diagnosed with psoriatic arthritis [PsA]). The discontinuation rate was 57.1%, the mean survival time of discontinuation was 57 weeks for patients with PsA vs 69 weeks for those without PsA (P = .5993). The cholesterol level (P = .003) and lymphocyte percentage (P = .010) were associated with longer survival of infliximab according to Cox regression analysis. Our study revealed that infliximab had a similar drug survival as previous studies, the high cholesterol level and lymphocyte percentage might function as negative predictor for infliximab persistence.

0025 Circadian Dysregulation of DNA Repair and Increased Endogenous and Exogenous Sensitivity to DNA Damage Precipitate Elevated Cancer Risk Associated with Night Shift Work

Abstract Introduction The elevated cancer risk associated with night shift work is poorly understood. To investigate whether circadian disruption may play a role, we assessed the circadian transcriptome and its association with hallmark cancer pathways, as well as sensitivity to endogenous and exogenous sources of DNA damage, after in-laboratory simulated shift work. Methods N=14 healthy humans (aged 22-34y; 10m, 4f) were exposed to a simulated night shift schedule (daytime sleep: 10:00-18:00) or a simulated day shift schedule (nighttime sleep: 22:00-06:00) for 3 days (n=7 in each condition). After the simulated shift schedule, subjects underwent a 24h constant routine protocol, during which blood was collected every 3h. Lymphocytes were extracted from the blood samples and subjected to transcriptome analysis using a NanoString multiplex assay. We evaluated 726 mRNA cancer hallmark targets (NanoString PanCancer Pathway Panel) and 17 circadian clock genes, with 18 arrhythmic internal controls. Gene expression was analyzed for circadian rhythmicity using mixed-effects cosinor analysis. Further, lymphocytes were investigated for DNA damage using an alkaline comet assay and immunofluorescence assessment of DNA damage response biomarkers BRCA1 and γH2AX. Lymphocytes collected at 07:30 and 19:30 were also exposed to ionizing radiation (2.5Gy) and DNA damage response assessments were repeated. Results Simulated night shift caused widespread disruption of circadian rhythmicity, as measured under constant routine, for core clock genes and the transcriptome of cancer hallmark pathways. The DNA repair pathway showed significant enrichment of rhythmic genes (p&amp;lt;0.05) after the simulated day shift schedule only. Following simulated night shift, lymphocytes showed induction of endogenous DNA damage, with extended tail in the comet assay (p&amp;lt;0.001), and higher percentage of lymphocytes with BRCA1 and γH2AX foci (p&amp;lt;0.01). Lymphocytes collected at 19:30 showed enhanced impact of ionizing radiation as indicated by increased prevalence of cells with BRCA1 and γH2AX foci (p&amp;lt;0.05). Conclusion Circadian dysregulation of DNA repair mechanisms and increased sensitivity to DNA damage following night shift work may increase genomic instability and precipitate elevated cancer risk in night shift workers. Support NIH grants ES022640 and CA227381, CDMRP award W81XWH-18-1-0100, and Pacific Northwest National Laboratory BRAVE investment under DOE contract DE-AC05-76RL01830.

Prognostic value of baseline complete blood count components in advanced gastric cancer patients: A multicenter retrospective study.

e16528 Background: Gastric cancer (GC) is one of the most prevalent malignant types in the world and an aggressive disease with a poor 5-year survival. Pretreatment CBC-based biomarkers, including blood neutrophil, lymphocyte, monocyte, and platelet counts; hemoglobin (Hb) levels; and their combinations, such as the neutrophil-lymphocyte ratio(NLR), lymphocyte-monocyte ratio (LMR) and platelet-lymphocyte ratio (PLR), have been reported to reflect systemic and local inflammation associated with cancer progression and prognosis. There has been growing interest in using CBC-based measures as biomarkers for GC. Methods: This chart-review study aimed to evaluate the effect of baseline levels of different components of routine CBC examination as well as other patients and disease characteristics on progression free survival (PFS) and overall survival (OS) in metastatic gastric cancer patients. Total 135 metastatic gastric cancer patients who had diagnosed and treated in three oncology centers in Saudi Arabia from 2011 to 2016 were incorporated. Various potential prognostic factors had measured in univariate and multivariate analysis. Results: After a median follow up of 21.4 months, the median OS / PFS were 11.0 and 6.1 months, respectively. Higher albumin level ( &gt; 3g/dl), low neutrophil percentage ≤ 75%, high lymphocyte percentage &gt; 15%, neutrophil /lymphocyte ≤ 2.5, high eosinophil count &gt; 0.4 k/ml, and EOX/EOF chemotherapy vs. doublet chemotherapy were associated with better PFS in univariate analysis. Conversely, in multivariate analysis, only serum albumin and eosinophil levels were related to PFS. In univariate analysis, higher serum albumin (3 g/dl), low neutrophil percentage ≤ 75%, high lymphocyte percentage &gt; 15%, neutrophil/lymphocyte ≤2.5, high eosinophil count &gt; 0.4 k/ml, receiving 1st line chemotherapy vs. no chemotherapy, receiving &gt; 6 cycles of chemotherapy, receiving EOX/EOF chemotherapy vs. doublet chemotherapy, platelet count ≤450 k/ml, male gender were associated with better OS. In multivariate analysis, lower neutrophil percentage, higher serum albumin, male sex and higher number of chemotherapy cycles were independently associated with OS. Conclusions: Higher eosinophil level was associated with improved PFS while lower neutrophil percentage and higher number of chemotherapy cycles were independent predictors of OS. Higher albumin levels independently predicted better OS and PFS.

Efficacy and safety of camrelizumab combined with apatinib in advanced triple-negative breast cancer: an open-label phase II trial

BackgroundPrevious trials showed that antiangiogenesis or anti-programmed death protein 1/programmed death ligand 1 (PD-1/PD-L1) monotherapy only showed marginal effect in triple-negative breast cancer (TNBC). Preclinical studies demonstrated that antiangiogenic therapy...

Correlation of bronchoalveolar lavage lymphocyte count with the extent of lung fibrosis and with plethysmographic lung volumes in patients with newly recognized hypersensitivity pneumonitis.

IntroductionHypersensitivity pneumonitis (HP) is an increasingly recognized interstitial lung disease, presenting with elevated total cell counts and high percentage of lymphocytes in bronchoalveolar lavage fluid (BALF). Despite many publications, there is no consensus in the literature concerning BALF cellular composition in patients with prolonged course of HP.AimThe aim of the present retrospective study was to investigate the influence of disease duration, smoking habits, and the extent of lung fibrosis on BALF cells’ population in patients with newly recognized HP.Material and methodsIn total, 94 patients (49 females, 45 males), mean age 52 (±12) years, with HP recognized according to recently proposed criteria, were enrolled into the present study. Chest CT scans were retrospectively reviewed by two independent radiologists. BALF evaluation was performed as a part of routine diagnostics according to recent recommendations.ResultsPercentage of lymphocytes in BALF was significantly lower in patients with lung fibrosis (stage 1 and 2) comparing to those without lung fibrosis (stage 0). Significant correlation was also found between the percentage of BALF lymphocytes and plethysmographic lung volumes, but not with lung transfer capacity for carbon monoxide (TLCO% pred). Smoking did not influence BALF results in our study group.ConclusionsBALF lymphocytosis correlated with the presence and the extent of lung fibrosis on chest CT as well as with plethysmographic lung volumes but did not with TLCO and smoking habits in newly recognized HP pneumonitis.

Examination of Factors Influencing the Success Rates of an In Vitro Chemosensitivity Test for Lung Cancer Using Collagen Gel Droplet-Embedded Cultures

Objective: We evaluated the success rates and factors influencing the success rates of the Collagen Gel Droplet-Embedded Culture Drug Sensitivity Test (CD-DST) for pleural effusate in lung cancer patients with carcinomatous pleurisy.Materials and methods: Thirty-two patients with lung cancer having carcinomatous pleurisy were enrolled in this study between January 2014 and May 2017. Their pleural effusions were analysed by CD-DST, and the subjects were classified into two groups: successful cases and unsuccessful cases, based on whether the test was able to successfully determine drug sensitivity. We investigated whether the properties of the pleural effusate might influence the success rate of CD-DST. We observed that the CD-DST tended to have good success rates in patients with higher pH, higher percentage of lymphocytes, higher cell counts, and lower percentage of neutrophils in their pleural effusate. The red blood cell count was higher in the effusate of unsuccessful than successful cases.Conclusion: Our results suggest that the diagnostic success rates of CD-DST for pleural effusion from patients with lung cancer might improve by appropriate sample selection.

Peripheral blood telomere alterations in ground glass opacity (GGO) lesions may suggest malignancy.

A ground glass opacity (GGO) lung lesion may represent early stage adenocarcinoma, which has an excellent prognosis upon prompt surgical resection. However, GGO lesions have broad differential diagnoses, including both benign and malignant lesions. Our objective was to study telomere length and telomerase activity in patients with suspected lung cancer in which GGO was the predominant radiographic feature. Knowledge of telomere biology may help distinguish malignant from benign radiographic lesions and guide risk assessment of these lesions. Peripheral blood samples were taken from 22 patients with suspected adenocarcinoma with the GGO radiographic presentation. Multidisciplinary discussion confirmed the need for surgery in all cases. We used an age and gender‐matched group without known lung disease as a control. Telomere length and aggregates were assessed by quantitative fluorescence in situ hybridization (QFISH) and quantitative PCR. Cell senescence was evaluated by senescence‐associated heterochromatin foci. Subjects with GGO lesions had a higher percentage of lymphocytes with shorter telomeres (Q‐FISH, P = 0.003). Furthermore, relative telomere length was also reduced among the GGO cases (qPCR, P < 0.05). Increased senescence was observed in the GGO group compared to controls (P < 0.001), with significant correlation between the senescence‐associated heterochromatin foci and aggregate formation (r = −0.7 and r = −0.44 for cases and controls, respectively). In conclusion, patients with resectable early adenocarcinoma demonstrate abnormal telomere length and cell senescence in peripheral blood leukocytes compared to control subjects. Abnormal telomere biology in the peripheral blood may increase suspicion of early adenocarcinoma among patients with GGO lesions.

Hematologic Markers as Prognostic Factors in Nonmetastatic Esophageal Cancer Patients under Concurrent Chemoradiotherapy.

Nonmetastatic esophageal cancer can demonstrate a high local recurrence rate even under the standard treatment. We evaluated platelet counts before and after concurrent chemoradiotherapy (CCRT), neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio for predicting esophageal cancer prognosis under CCRT. Newly diagnosed patients with esophageal cancer (stages IA–IIIC) who underwent CCRT during January 2013–December 2017 were enrolled. The data were collected retrospectively. Overall survival (OS), time to progressive disease (TPD), and time to metastasis (TM) were recorded for indicating prognosis. Kaplan–Meier curves were plotted and univariate and multivariate analyses were performed. In total, 105 patients were enrolled. The stages of esophageal cancer and surgery were associated with prognosis (i.e., OS, TPD, and TM). Based on TPD and TM, women had better prognosis than men. In the univariate analysis, high pre- and post-CCRT platelet counts (>300,000/μL), platelet-to-lymphocyte ratio (PLR), and neutrophil-to-lymphocyte ratio (NLR) as well as low lymphocyte percentage were significantly associated with poor prognosis. However, in the multivariate analysis, only post-CCRT high platelet count (>300,000/μL) remained significantly associated with poor prognosis (P = .041, .045, and .023 for OS, TPD, and TM, respectively). Poor prognosis was observed in patients with high platelet counts, PLR, NLR, and low lymphocyte percentage. Surgery was an independent factor predicting better prognosis. Our findings may have clinical significance with regard to therapeutic decision-making.

Insulin Rescues Diabetes-Induced Pulmonary Complications

Hyperglycemia is an independent risk factor for the development of respiratory diseases, including influenza infections. Although the lung is a major organ to utilize glucose, the regulation of glucose transport in the healthy and diabetic lung has received little attention. We hypothesized that hyperglycemia would predispose diabetic mice to alterations in pulmonary glucose transport and pulmonary complications during Influenza A infection. To test this hypothesis, we used a streptozotocin-induced type 1 diabetic mouse model (n=8-12/group). A subset of diabetic mice was treated with insulin via a subcutaneous insulin pump to restore euglycemia. After 8 weeks, an additional subset of mice was intranasally infected with influenza A H1N1 (A/PR/8/34; 250 PFU). Viral titer was determined from bronchoalveolar lavage (BAL) fluid by plaque assay. Glucose concentration in BAL fluid was measured spectrophotometrically. Glucose transporter (GLUT) protein content was quantified in whole lung homogenates by Western blotting. Both infected and uninfected diabetic mice had higher glucose concentrations in BAL fluid than their control or insulin-treated counterparts (p&amp;lt;0.05). There was a trend towards a decrease in protein content of GLUT4 (the major insulin-sensitive isoform) and GLUT1 (a basal isoform) in the diabetic lung (p&amp;lt;0.1). Total protein content of GLUT12 (a novel isoform) was increased in the diabetic lung (p&amp;lt;0.05). Diabetic mice possessed a significantly higher percentage of lymphocytes in their BAL fluid (p&amp;lt;0.05). GLUT protein content and lymphocyte alterations were rescued when mice were treated with insulin. Influenza-infected diabetic mice also had a higher percentage of lymphocytes (p&amp;lt;0.05), and a trend toward an increase in viral load in BAL fluid vs. infected control mice. These novel findings suggest that 1) the regulation of glucose transport is altered in the lung during hyperglycemia, and 2) diabetic mice are predisposed to pulmonary inflammation and viral replication during influenza infection. Disclosure A. Campolo: None. Z. Maria: None. M. Hinsdale: None. L. Liu: None. V. Lacombe: None.

HEMATOLOGY AND SERUM BIOCHEMISTRY VALUES OF THE RED PANDA SUBSPECIES ( AILURUS FULGENS STYANI).

Health evaluations were conducted on individuals ( n = 62, age < 1 to 11 yr) from a captive red panda ( Ailurus fulgens styani) population at the Chengdu Research Base for Giant Panda Breeding in Sichuan Province, People's Republic of China. Individuals were anesthetized using combinations of ketamine, dexmedetomidine, diazepam, isoflurane, and tiletamine/zolazepam. Whereas physical exams revealed no abnormalities for 40% of red pandas, 24% had dental abnormalities and 18% had orthopedic abnormalities. As the red panda is the only member of the family Ailuridae, complete blood count and serum biochemistry samples were collected and used to develop reference intervals for this species. When hematologic and serum chemistry values of male and female red pandas were compared within this A. f. styani population, males had higher mean cell hemoglobin, mean cell hemoglobin concentration, and blood urea nitrogen concentrations, whereas females had higher alkaline phosphatase, cholesterol, and iron. Adult red pandas (≥2 yr) had higher lymphocyte percentage, total protein, globulin, and chloride levels, whereas juveniles (<2 yr) had higher white blood cell concentrations, neutrophil percentage and absolute neutrophil values, alkaline phosphatase, creatine kinase, cholesterol, potassium, total and ionized calcium, and phosphorus levels. Reference values and expected age and sex differences will assist with health management of the captive A. f. styani population.

Dietary Bovine Lactoferrin Reduces Staphylococcus aureus in the Tissues and Modulates the Immune Response in Piglets Systemically Infected with S. aureus

BackgroundBovine lactoferrin (bLf) reduces Staphylococcus aureus infection in premature infants and promotes the growth of Bifidobacterium infantis, a predominant infant gut species. We hypothesized that bLf in combination with B. infantis would reduce the severity of systemic S. aureus infection. ObjectiveThe aim was to determine the effects of oral administration of bLf and B. infantis on the course of systemic S. aureus infection. MethodsColostrum-deprived piglets were fed formulas containing 4 g whey/L (CON group) or bLf (LF group). One-half of the piglets in each group were gavaged with B. infantis (109 colony-forming units/d), resulting in 2 additional groups (BI or COMB, respectively). On day 7, piglets were intravenously injected with S. aureus. Blood samples were collected preinfection and every 12 h postinfection for immune analyses. Tissue samples were collected on day 12 for analysis of bacterial abundance and gene expression. ResultsPreinfection, LF piglets had lower serum interleukin 10 (IL-10), a higher percentage of lymphocytes, and a lower percentage of neutrophils than BI or COMB piglets. After infection, dietary bLf increased piglet weight gain, reduced staphylococcal counts in the kidneys, and tended to lower staphylococcal counts in the lungs and heart. Dietary bLf also decreased kidney IL-10 and increased lung interferon γ (IFN-γ) mRNA. B. infantis increased splenic IFN-γ expression. Renal Toll-like receptor 2 was upregulated in BI piglets but not in COMB piglets. Postinfection, BI piglets had increased serum IL-10 and decreased memory T cell populations. LF and COMB piglets had fewer circulating monocytes and B cells than CON or BI piglets. ConclusionsDietary bLf and B. infantis produced independent and tissue-specific effects. Piglets fed bLf alone or in combination with B. infantis mounted a more effective immune response and exhibited lower bacterial abundance. This study provides biological underpinnings to the clinical benefits of bLf observed in preterm infants but does not support B. infantis administration during S. aureus infection.

Dynamics of morphological and biochemical indicators of blood of sucking piglets at different levels of iodine citrate in sows rations

The article presents the results of experimental studies on the determination of the influence of iodine citrate on hematological and biochemical parameters of blood of sucking piglets. It was established that hemopoiesis and metabolic reactions in pigs depend on the level of Iodine in the sows, and in the later period (the 10th day) and from the receipt of the biometal as part of the prestarter. Iodine, in nanocitate form, at the expense of high bioavailability and chemical activity, provided the need for thyroid gland in it in much less quantities than the amounts recommended for pregnant and lactating sows in the form of potassium iodide. It has been established that at the admission to the body of a sow 0.095–0.19 mg/kg of feeding of iodine in a nanocyte form (1/4–1/2 of the recommended amount of iodine in the form of CI), its positive effect on hemopoiesis and metabolic processes in newborn piglets is peculiar (increased number of red blood cells, hemoglobin content, high percentage of lymphocytes, increases in the concentration of total protein and in particular albumin, against the backdrop of growth in blood plasma levels of CSF, the concentration of hormones T3 and T4 is quite stable. A higher dose in the premix of iodine citrate (1 : 1) is irrational and most likely undesirable, since against the background of inhibition of this group of erythropoiesis, eosinophilia and lymphopenia, the protein synthesis function decreases and the activity of AlAT, AsAT increases. Hematological and biochemical parameters of blood of piglets obtained from sows, which received a minimum amount of iodine in the form of citrate (0.1 : 1) with premix, did not undergo significant changes, were within the limits of physiological values, although in comparison with the animals of the control group, and particularly, the piglets of the group D2 and D3 tended to decrease (growth). Consequently, iodine citrate as a source of iodine for pregnant and subsistence sows can be recommended in the amounts: 0.095–0.19 mg/kg and 0.125–0.25 mg/kg feed.

Surface CD200 and CD200R antigens on lymphocytes in advanced gastric cancer: a new potential target for immunotherapy.

IntroductionGastric cancer (GC) is one of the leading causes of cancer death worldwide. The membrane glycoprotein CD200, widely expressed on multiple cells/tissues, uses a structurally similar receptor (CD200R), delivering immunoregulatory signals. There is evidence that CD200/CD200R signaling suppresses anti-tumor responses in different types of malignancies. Little is known about the CD200/CD200R pathway in GC. The aim of the study was to evaluate the frequencies of CD200+ and CD200R+ lymphocytes in patients with GC.Material and methodsForty patients primarily diagnosed with GC and 20 healthy volunteers (control group) were enrolled. The viable peripheral blood lymphocytes underwent labeling with fluorochrome-conjugated monoclonal antibodies and were analyzed using a flow cytometer.ResultsIn the GC group, the percentages of T CD3+, CD3+/CD4+, and CD3+/CD8+ cells expressing CD200 antigen were higher than in the control group (p < 0.00013, p < 0.0004, and p < 0.0006, respectively). In the GC group, the frequencies of T CD3+, CD3+/CD4+ and CD3+/CD8+ cells expressing CD200R were lower than in the control group (p < 0.0009, p < 0.004, and p < 0.002, respectively). The percentage of B CD19+/CD200+ lymphocytes was higher in GC patients than in the control group (p < 0.00005). Lower frequency of B CD19+/CD200R+ cells was observed in GC patients compared to the control group (p < 0.0001). No differences in the frequencies of CD200+ and CD200R+ lymphocytes were found in relation to either UICC stage or histological grading of the tumors.ConclusionsFor GC pathogenesis, deregulation of the CD200/CD200R axis is important. High percentages of lymphocytes with CD200 expression may contribute to the continuous T cell activation and development of chronic inflammation and influence gastric carcinogenesis.

The PD-1/PD-L1 Inhibitory Pathway is Altered in Primary Glomerulonephritides

The pathogenesis of primary proliferative and non-proliferative glomerulonephritides (PGN and NPGN) is still not fully understood, however, current evidence suggests that most cases of PGN and NPGN are the results of immunologic response to different etiologic agents that activates various biological processes leading to glomerular inflammation and injury. Programmed cell death protein 1 (PD-1) is the major inhibitory receptor regulating T cell exhaustion. The aim of this study was to evaluate the frequencies of PD-1-positive and PD-ligand 1 (PD-L1)-positive T and B lymphocytes in patients with NPGN and PGN in relation to clinical parameters for the first time. The study included peripheral blood (PB) samples from 20 newly diagnosed PGN and NPGN patients. The control group comprised of 20 healthy age- and sex-matched subjects. The viable PB lymphocytes underwent labelling with fluorochrome-conjugated monoclonal antibodies anti-PD-1 and anti-PD-L1, and were analyzed using a flow cytometer. The frequencies of CD4+/PD1+ T lymphocytes, CD8+/PD1+ T lymphocytes, and CD19+/PD-1+ B lymphocytes in the PGN group exceeded values obtained both in the NPGN group, and the control group. Alteration of PD-1/PD-L1 pathway may be involved in poorer prognosis, as patients with PGN are characterized by higher frequencies of PD-1-positive and PD-L1-positive T and B lymphocytes than patients with NPGN. Our results suggest that deregulation of PD-1/PD-L1 axis may contribute to the PGN and NPGN pathogenesis. High percentages of lymphocytes with PD-1 and PD-L1 expression may be related to the continuous T-cell activation and development of glomerular inflammation and injury.