Absolute Number Of Lymphocytes Research Articles

Cyclosporin A and doxorubicin-ifosfamide in resistant solid tumours: a phase I and an immunological study

In order to test whether circumvention of clinical resistance can be obtained in common solid tumours by targeting different drug resistance mechanisms, a phase I clinical and immunological study was designed. The purpose of the study was to determine the dose of cyclosporin A (CsA), in combination with doxorubicin (DOX) and ifosfamide (IFX), needed to achieve steady-state whole-blood levels of 2000 ng ml-1 and the associated toxicity of this combination. Treatment consisted of CsA 5 mg kg-1 as a 2 h loading infusion, followed by a CsA 3 day continuous infusion (c.i.) (days 1-3) at doses that were escalated from 10 to 18 mg kg-1 day-1. Chemotherapy consisted of DOX 55 mg m-2 by i.v. 24 h c.i. (day 2) and IFX 2 g m-2 i.v. over 1 h on days 1 and 3. Treatments were repeated every 4 weeks. Eighteen patients with previously treated resistant solid tumours received 39 cycles. Mean steady-state CsA levels > or = 2000 ng ml-1 were reached at 5 mg kg-1 loading dose followed by a 3 day c.i. of 16 mg kg-1 day-1 or greater. Haematological toxicity was greater than expected for the same chemotherapy alone. One patient died of intracranial haemorrhage due to severe thrombopenia. Other observed toxicities were: asymptomatic hyperbilirubinaemia (46% cycles), mild nephrotoxicity (20% cycles), hypomagnesaemia (72% cycles), mild increase in body weight (100% cycles), hypertension (15% cycles) and headache (15% cycles). Overall the toxicity was acceptable and manageable. No alterations in absolute lymphocyte number, the lymphocyte subsets studied (CD3, CD4, CD8, CD19) or CD4/CD8 ratio were observed in patients receiving more than one treatment cycle, although there were significant and non-uniform variations in the values of the different lymphocyte subsets studied when pre- and post-treatment values were compared. There was also a significant increase in the CD4/CD8 ratio. Tumour regressions were observed in two patients (epidermoid carcinoma of the cervix and Ewing's sarcoma). The CsA dose recommended for phase II trials is a 5 mg kg-1 loading dose followed by a 3-day c.i. of 16 mg kg-1 day-1 simultaneously with DOX and IFX at the doses administered in this study.

Lymphocytes with Fas-receptors of readiness to apoptosis in non-psychotic mental disorders

IntroductionApoptosis is a complex physiological process of the organism which supports cellular homeostasis, provides important aspects of development and functioning of the immune system. In various pathological conditions the process of apoptosis can be impaired that leads to decrease or increase in pro-apoptotic activity.Materials and methodsWe conducted investigation of relative and absolute number of CD3+CD95+–lymphocytes in groups of patients with adjustment disorders (n = 90), PTSD (n = 100), organic emotionally labile (asthenic) disorder (n = 232), organic personality disorder (n = 93). Clinical verification was conducted according to ICD–10. Control group included 190 practically healthy persons. Fas protein (CD95) expression on CD3 lymphocytes surfaces was detected using flow cytometry. Cytometric measurements were conducted on flow cytofluorimeter FacsCalibur (Becton Dickinson, US).ResultsIn the control group relative number of CD95+–lymphocytes was 11.6%, absolute–0.21 × 109/L. In all examined patients as compared with control the reliable increase both in relative and absolute number of lymphocytes of CD3+CD95+–phenotype was identified. So, in persons with adjustment disorder content of this indicator made 17.0% and 0.28 × 109/L (Р = 0.0015), in PTSD–18.0% and 0.33 × 109/L (Р = 0.0007) and in patients with organic asthenic disorder–19.0% and 0.32 × 109/L (Р = 0.0048), respectively. The highest content in blood of CD3+–lymphocytes, expressing on the surface of membrane the basic marker of apoptosis CD95 is observed in patients with organic personality disorder: 26.0% and 0.44 × 109/L (Р = 0.0003).ConclusionIn case of intensification of psychopathological symptoms especially in persons with non-psychotic organic mental disorders a receptor-mediated signaling pathway of apoptosis is activated – process of programmed cell death.Disclosure of interestThe authors have not supplied their declaration of competing interest.

Evaluation of blood T-lymphocyte subpopulations involved in host cellular immunity in dogs with mammary cancer

Cancer-bearing patients are often immunosuppressed. In dogs with mammary or other cancers, various alterations in blood cell populations involved in host cellular immunity have been reported; among these cell populations some T-lymphocyte subsets play an important role against cancer. The purpose of the present study was to investigate any alterations in circulating T-lymphocyte subpopulations involved in cellular immunity in bitches with mammary cancer, in comparison to age-matched healthy intact bitches. Twenty eight dogs with mammary cancer and 14 control dogs were included in this study. Twelve out of the 28 bitches had mammary cancer of clinical stage II and 16/28 of stage III. Histological examination revealed that 23/28 animals had carcinomas, 3/28 sarcomas and 2/28 carcinosarcomas. White blood cell, neutrophil and lymphocyte absolute numbers were measured by complete blood count. Furthermore, blood T-lymphocyte population (CD3+) and the subpopulations CD4+, CD8+ and CD5low+ were assessed by flow cytometry. White blood cell and neutrophil but not lymphocyte absolute numbers were higher (P=0.003 and P=0.001, respectively) in cancer patients than controls. Flow cytometric analysis revealed that the relative percentage of T-lymphocytes (CD3+) and of CD4+, CD8+ subpopulations was lower (the CD4+/CD8+ ratio was higher), whereas the percentage of CD5low+ T-cells was higher, in dogs with cancer compared to controls; however, a statistically significant difference was found only in the case of CD8+ T-cells (P=0.014), whereas in the case of the CD4+/CD8+ ratio the difference almost reached statistical significance (P=0.059). Based on these findings, it can be suggested that, although the absolute number of blood lymphocytes is unchanged, the relative percentages of T-lymphocyte subpopulations involved in host cell-mediated immunity are altered, but only cytotoxic CD8+ T-cells are significantly suppressed, in dogs with mammary cancer of clinical stage II or III compared to age-matched healthy controls.

ASP0028 in combination with suboptimal-dose of tacrolimus in Cynomolgus monkey renal transplantation model

FTY720, a S1P-receptor modulator, has shown to be effective in several transplant and autoimmune disease models, via modulating lymphocyte homing into secondary lymphoid organs (SLOs), and thereby reducing these cells in peripheral blood. ASP0028, a newly developed S1P1/S1P5-selective agonist, presented comparable efficacy to FTY720 and wider safety margins than FTY720. In this study, we assessed the efficacy and safety of ASP0028 co-administered with suboptimal-dose of tacrolimus in the Cynomolgus monkey renal transplantation model. Seven animals in group-1 or group-2 received mono-tacrolimus 1.0mg/kg once a day (QD), or ASP0028 0.6mg/kg plus tacrolimus 1.0mg/kg QD, respectively. Eight animals in group-3 received ASP0028 1.2mg/kg plus tacrolimus 1.0mg/kg QD. The allograft median survival time (MST) in group-2 and group-3 were significantly extended to 41 and 61.5days, versus that of 28days in group-1 (p=0.036 and 0.001, respectively). ASP0028 administration remarkably reduced absolute numbers of peripheral lymphocytes, particularly subsets of CD4+/ or CD8+/naive and central memory cells, CD4+/Treg cells, and to a lesser extent on B cells, but not CD4+/ or CD8+/effector memory cells and NK cells. These data show ASP0028 combined with suboptimal-dose of tacrolimus effectively prolongs renal allograft survival in nonhuman primates (NHPs) with well tolerated safety, supporting its further investigation to optimize CNI-sparing regimens.

STUDY OF SOME IMMUNOLOGICAL AND HEMATOLOGICAL IMPACTS INDUCED BY CLOMAZONE IN SWISS ALBINO MICE AND THE PROTECTIVE EFFECT OF VITAMIN C

ABSTRACT: The present study aimed to evaluate the probable perturbations produced by herbicide clomazone (CMZ) on immunological and hematological indices and to assess the protective role of vitamin C in alleviating CMZ-induced immunological and hematological toxicity in male Swiss albino mice. To achieve this goal, Swiss albino mice (weighing 25-34 g and each group 10 animals) were administrated phosphate buffer saline (PBS), CMZ (46 µM/kg; 1/20 of the 96h LD50 for an intraperitoneal dose) and/or vitamin C (1136 µM/kg) daily intraperitoneally (i.p.) for 4 weeks. By the end of the 4th week, immunological and hematological parameters: erythrocyte (RBC) count, hemoglobin (Hb) percent, hematocrit (Hct) content, platelets (Plt) count, leucocyte (WBC) count, absolute number of neutrophils, lymphocyte and monocytes and count of splenocytes, thymocytes and bone marrow cells were evaluated. To investigate the influence of immunoglobulin (Ig) isotypes production activity of CMZ, naive splenocytes were inoculated for 24 h into media supplemented with PBS, CMZ (4.6 µM/well) and/or vitamin C (113.6 µM/well) and the concentrations of IgA, IgG and IgM were measured by ELISA. As a result, 96h LD50 of CMZ was estimated at 911 µM/kg. Exposure to CMZ alone significantly decreased RBC count, Hb content, Hct%, WBC count, absolute numbers of lymphocytes and monocytes, Plt count, total numbers of splenocytes, thymocytes and bone marrow cells when compared to their values in PBS-treated mice. Contrary, Absolute number of neutrophils was significantly higher in CMZ-treated mice than in PBS-treated mice. The concentrations of IgA, IgG and IgM in CMZ-cultured splenocytes were significantly higher than in PBS-cultured splenocytes. Pre-treatment of vitamin C to CMZ-exposed mice mildly ameliorated the immunological and hematological perturbations of CMZ toxicity. To conclude, it appears that vitamins C mildly alleviated CMZ–induced immunological and hematological toxicity but it was not completely protective.

КЛИНИКО-ИММУНОЛОГИЧЕСКИЕ ОСОБЕННОСТИ ОСТРОНЕКРОТИЗИРУЮЩЕЙСЯ ФОРМЫ КОЖНОГО ЛЕЙШМАНИОЗА

Objective: To study immunological disorders in patients with a rural (acute necrotizing) form of Cutaneous Leishmaniasis (CL). Methods: 80 patients were found under the observation in a rural (acute necrotizing) form of CL, including 23 (28.8%) men and 57 (71.2%) of women. The age of patients fluctuated from 16 to 60 years and over Results: The data showed that the oppression of the activity of the cellular immunity in patients with CL in comparison with healthy people, which indicates the significant of absolute and comparative numbers of CD3 lymphocytes (by 20%, p 0.01), the tendency of falling of absolute and comparative numbers of CD4 (T helper-inducers) and CD8 (T-cytotoxic) lymphocytes (correspondingly to 12% and 18%), as well as the absolute and comparative number of lymphocytes (by 12%) of peripheral blood. Conclusion: Thereby, in patients with acute necrotizing form of CL observes immunological disorders both in cellular and humoral components of immunity, which was depended on prescription of the process and the presence of complications. Keywords: Acute necrotizing cutaneous leishmaniasis, clinical and immunological features.

The Novel Patient with BLNK Gene Type of Agammaglobulinemia

Background: Agammaglobulinemia (AGM) is a genetic immune system disorder in which the body could not produce antibodies. There are two types of this disease: X-linked AGM (XLA) and autosomal forms. X-linked agammaglobulinemia results from a mutation in the gene for Bruton’s tyrosine kinase (BTK), found on the X chromosome. Mutations in BTK result in lack of mature B-cells and immunoglobulins of all classes. There are 8 types of autosomal forms which are reflected in different defective genes. 7-year-old male patient at the age of 1-6, was encountered the recurrent broncho-pulmonary and gastro-intestinal infections, furunculosis and recurrent otitis. Also, because of the strain, pain and swelling on the 4th finger of the right hand it was suspected that the patient had rheumatoid arthritis. On immunological investigation, we have found out of absent of B lymphocyte. Level of IgG was 10 times less; IgA, IgM and IgE levels were 3 to 8 times less than the norm. Quantity of T-lymphocytes: absolute number of CD4 lymphocytes increased, CD3 and CD8 lymphocytes were also high. Phagocyte activity in NBT, absolute number of NK cells and IRI index were 2 times lower than the norm. Genetically analysis has discovered the presence of abnormal homozygous BLNK gene.

CD14-CD15+, Possible Cells Are Potent Myeloid-Derived Suppressor Cells, Which Affect the Pathogenesis of Induction and Regression in Methotrexate-Mediated Lymphoproliferative Disorders

[Background]Despite numerous attempts to elucidate the mechanism underlying other iatrogenic immunodeficiency-associated lymphoproliferative diseases (OIIA-LPDs), the mechanism remains poorly understood. Methotrexate-associated lymphoproliferative disorder (MTX-LPD) is one of the disease entities included in OIIA-LPDs, which is especially developed in rheumatoid arthritis (RA) as we demonstrated (Tokuhira et al, Leuk Lymphoma. 2012;53:616-23). Spontaneous regression of LPD after MTX withdrawal is one of the distinct characteristics of MTX-LPD, and recent articles suggest that the increase in peripheral blood (PB) lymphocytes affect this phenomenon. However, the population of lymphocytes necessary to induce MTX-LPD regression is still unknown.[Methods]We prospectively analyzed 10 patients with RA developing confirmed LPD during MTX administration, in our institutions between January 2014 to October 2015. All patients ceased MTX after developing lymphoadenopathies. To investigate the factors involved in spontaneous regression of LPD following MTX withdrawal, we performed flowcytometric analysis of whole blood including lymphocytes from PB for CD3, 4, 8, 14, 15, 20, 56, 127, and 45RO, HLA-DR, CCR3, CCR4, CCR6, CCR7, and EBV-tetramer expression, for all patients at three time points during the clinical course of treatment; at the time of MTX cessation (w0), and 4 weeks (w4) and 12 weeks (w12) after MTX cessation. We selected 10 age-, sex-, MTX dose-, and RA duration-matched RA patients treated with MTX for more than 6 months as controls, randomly selected from among the RA patients at our institutions. The patients with MTX-LPD were divided into two groups based on the status of LPD after MTX cessation, regressive LPD (R group) and persistent LPD (P group), and clinical parameters were compared.[Results]In the 10 RA patients with MTX-LPD, the median age was 62.4 years (40-74), and the ratio of male:female patients was 2:8. The median duration from the time of RA diagnosis to the time of MTX development was 7.2 years (1.2-20.4), and the median duration of MTX administration was 4.3 years (0.7-9.8). The pathological diagnosis of LPDs was diffuse large B cell lymphoma (DLBCL, n=5), Classical Hodgkin lymphoma (cHL, n = 3), and non-specific LPD (LPD-nos, n=2). The averages values for LDH, CRP, and sIL-2R in the serum were 220 IU/mL (176-554), 0.85 mg/dL (0.06-2.53), and 579 IU/L(206-1210), respectively. The number of patients in the R group and the P group was 7 and 3, respectively. According to LPD phenotypes, all 5 cases of DLBCL belonged to the R group, whereas 1 of 3 HL and 1 of 2 LPD-nos belonged to the R group.At w0, the median WBC count and absolute lymphocyte number in 10 MTX-LPD patients were 5810/µL (2700-11400) and 964/µL (220-2070), respectively. On the other hand, the median WBC count and absolute lymphocyte number in the 10 control patients were 5930/µL (3900-9000) and 1512/µL (755-2379), respectively. The absolute lymphocyte count in the P group was significantly higher than that in the R group. In addition, a significant increase in the lymphocyte number following MTX withdrawal was observed only in the R group, but not in the P group.There were several observations from the flowcytometric analysis. First, the proportions of effector memory CD8+ T cells (EM CD8+), EBV specific CD8+ T cells, and T helper 1 (Th1) cells were significantly decreased both in the R group and in the P group compared to the control group at w0, and this subset was significantly increased in the R group at w4 and w12 compared to the P group. Second, the proportion of CD14-CD15+ cells in the lymphocyte fraction, which includes myeloid-derived suppressor cells (MDSC), was significantly increased in LPD group at w0, and significantly decreased following MTX cessation only in the R group, but not in the P group. Third, the proportion of CD14-CD15+ cells negatively and significantly correlated with the proportion of EM CD8+T cells (r2 = 0.47, p < 0.0001).[Conclusions]Although the mechanism of MTX-LPD regression following MTX withdrawal is still unknown, this study demonstrates that reconstitution with specific lymphocytes in the PB is definitely correlated with LPD-regression. The proportion of Th1 cells, EM CD 8+, EBV specific CD8+ along with CD14-CD15+ cells dramatically indicated restoration and transition following MTX cessation, suggesting their potent effect toward the progression and regression of MTX-LPD. DisclosuresTokuhira:Bristol Myers Squibb Co., Ltd: Honoraria; Pfizer Co., Ltd: Honoraria; Eizai Co., Ltd: Honoraria. Okamoto:Otsuka Pharmaceutical Co., Ltd.: Honoraria, Research Funding; Shionogi & Co., Ltd.: Research Funding; Sumitomo Dainippon Pharma Co., Ltd.: Research Funding; Kyowa Hakko Kirin Co., Ltd.: Research Funding; Eisai Co., Ltd.: Research Funding; Teijin Pharma Limited: Research Funding; Chugai Pharmaceutical Co., Ltd.: Research Funding; Toyama Chemical Co., Ltd.: Research Funding; Nippon Shinyaku Co., Ltd.: Research Funding; Bristol-Myers Squibb K.K.: Honoraria, Research Funding; Asahi Kasei Pharma Corp.: Research Funding; Astellas Pharma Inc.: Research Funding; Alexion Pharmaceuticals, Inc.: Research Funding; Pfizer Inc.: Honoraria, Research Funding; JCR Pharmaceuticals Co., Ltd.: Research Funding.

Adult Patients with Acquired Pure Red Cell Aplasia: Treated By Cyclosporine a and/or Corticosteroids--Single Center Experience

▪Objective: Adult pure red cell aplasia (PRCA) is a syndrome characterized by a severe normocytic anemia, reticulocytopenia, and absence of erythroblasts from an otherwise normal bone marrow. Immunosuppressive therapy has been used as the initial treatment for acquired chronic PRCA. This article aims to evaluate the efficacy of cyclosporine A, and/or corticosteroids, and possible factors influencing it.Methods: 34 cases of PRCA were retrospectively analyzed at our institution.Clinical data of 23 inpatient cases and 11 outpatient cases from 2009 October to 2016 March were collected. These patients were treated by cyclosporine A (CsA), and/or corticosteroids (CS).Results: The causes of PRCA were as follows (table 1): idiopathic (15 cases, 44.1%), thymoma-associated (5 cases, 14.7%), and large granular lymphocyte (LGL) leukaemia-associated (7 cases, 20.6%), virus infection (4 cases, 11.7%), major ABO-mismatched allogeneic haematopoietic stem cell transplantation (1 case, 2.94%), rheumatoid arthritis (1 case, 2.94%), antierythropoietin antibody-mediated (1 case, 2.94%). One patient was lost to follow-up. Two patients were not evaluable for response due to short observation period (<3 months). In idiopathic PCRA, complete remission (CR) was achieved in 4 of 14 cases (28.6%), and partial remission (PR) was achieved in 7 patients (50.0%), while 1 case (7.1%) was still transfusion dependent and 2 deaths(14.3%). In secondary PRCA, complete remission (CR) was achieved in 6 of 17 cases (35.3%, 3 LGL, 2 thymoma, 1 others), and partial remission (PR) was achieved in 3 patients (17.6%, 1 LGL, 2 others ), while 7 case (41.2%, 3 LGL, 2 thymoma, 2 others)) were still transfusion dependent and 1 death(5.9%, thymoma). It showed that the initial rate of CR in CsA-containing group was higher than CS alone (33.3% vs 0%, P=0.028). In 22 refractory and relapsed PRCA patients, 6 out of 10 (60%) refractory patients and 5 out of 12 (41.7%) relapsed patients achieved remission(table 2). The cumulative rate of CR of CsA alone, CS alone, CS+CsA and other immunosuppressive agents were 27.8%, 6.67%, 66.7%, 10%. The cumulative rate of CR of the combination of CS and CsA was higher than CS alone or CsA alone (66.7% vs 18.2%, P=0.028). The response rate of CS or CsA was higher than other immunosuppressive agents (69.2% vs 20%, P<0.01). The age, gender, the percent of reticulocyte and absolute number of lymphocyte in peripheral blood, and etiology did not influence the response rate. (table 3)Conclusion: Thymoma and LGL are the two common causes of secondary PRCA. CsA and/or CS are effective in treating PRCA.The combination of CS and CsA was more effective in achieving CR than CS alone or CsA alone. It was suggested that we might choose the therapy which contains CsA as the initial induction therapy. Other immunosuppressive agents (tacrolimus or mycophenolate mofetil) was less effective in refractory and relapsed PRCA patients. And it was still needed to explore a more effective therapy for refractory and relapsed PRCA patients. [Display omitted] [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

One-Time Administration of Recombinant Human Erythropoietin Decreases B Cell in Human Peripheral Blood

Introduction:Erythropoietin (EPO) is hematopoietic factors participating in red blood cell production by accelerating cell proliferation and inhibition of apoptosis of erythroblast. It is reported that EPO has pleiotropic effects including anti-apoptotic action for some cells, antioxidant action, vascularization action, and promoting cell division in addition to stimulation of erythropoiesis as well, whereas there are conflicting results of small cohorts as to its effect on blood immune cells. We analyzed peripheral white blood cell subsets in patients who received one bolus administration of recombinant human erythropoietin (rHuEPO) to examine the effect of EPO on human immune system.Materials and Methods:One hundred twenty eight autologous blood donors (male/female 64/64) in Gunma University Hospital were enrolled in this study after written informed consent. All the patients had no infections or inflammation. Fifty six patients were administered rHuEPO (Epoetin alpha or Epoetin beta, 24,000 IU) once after blood donation because of low hemoglobin concentration (EPO group) and 72 were not administered (non-EPO group). Peripheral blood samples were obtained at the time of the first phlebotomy and after 7 days from the same patient.We measured the number of WBC, lymphocytes, myeloid dendritic cells (mDC), plasmacytoid dendritic cells (pDC), CD4+ T cells, CD8+ T cells, Natural killer (NK) cells, B cells, monocytes, and neutrophils of peripheral blood before and after rHuEPO administration by flow cytometry. Also we compared EPO group with non-EPO group. Paired and unpaired Student’s t-test were used to compare absolute counts and percentages of each cell, P values < 0.05 were considered significant. This study was approved by the research ethics committee of our hospital.Results:In EPO group, absolute number and percentage of lymphocyte in WBC decreased significantly after rHuEPO administration from 1754.9 ± 535.4/µl to 1627.2 ± 467.7/µl, (p<0.01) and from 30.9 ± 8.00% to 28.2 ± 6.77% (p<0.01), respectively. The numbers of whole WBC, mDC, pDC, monocyte and neutrophil did not show significant changes. In regards to lymphocyte subset, absolute number of CD8+ T cell significantly decreased from 358.9 ± 257.0/µl to 311.5 ± 210.9/µl (p=0.04) and that of NK cell from 290.6 ± 157.6/µl to 257.4 ± 141.8/µl (p<0.01), but their percentages did not change. B cells significantly decreased in absolute number from 258.3 ± 154.2 /µl to 221.6 ± 129.2 /µl (p<0.01) and also in percentages from 14.8 ± 6.92% to 13.6 ± 6.07% (p<0.01). Regarding B cell subsets, absolute number of Naïve B cells significantly decreased from 171.3 ± 93.5 /µl to 153.0 ± 84.2 /µl (p<0.01); moreover other B cell subsets, such as transitional B cells, memory B cells and marginal zone B cells, also showed a trend of decrease, so that the percentage of Naïve B cell did not change in total B cells. These suggested that whole B cell decreased, not a specific subset of B cell. In non-EPO group, there was no significant change in any type of immune cells. When compared the rate of change between before and after rHuEPO treatment, B cell number significant decreased in EPO group compared to non-EPO group (0.87 vs 0.98, p<0.01). Multiple regression analysis revealed rHuEPO administration significantly influenced B cell number (p=0.015); however age, sex, disease status and the type of rHuEPO preparations (epoetin alfa or beta) did not influence.Conclusion:These findings suggested that just one administration of rHuEPO influenced human immune system, especially via reduction of B cell in peripheral blood. DisclosuresNo relevant conflicts of interest to declare.

IMPACT OF EMOTIONAL STRESS ON THE IMMUNE SYSTEM INDICES AMONG RESIDENTS OF RADIATION CONTAMINATED AREAS.

The aim of this research is to study special characteristics of immune system functioning among residents of the areas contaminated with radionuclides, under conditions of additional emotional load. We examined 350 people, including a group of radiation free areas residents (control group, 150 people) and a group of residents living in the areas of enhanced radiation monitoring (IV radiation zone, the density of soil contamination with isotopes (137)Cs 1-5 Ki/km(2), 200 people). All examined are the students of Cherkassy State University, aged 18-24 years, at the time of research didn't have any acute diseases. The role of additional stress factor (additional emotional load) was fulfilled by winter examination session. The first analysis of immune system indices was carried out in the interval between examination sessions, the second 2 after the first exam, the third - after the last exam, the fourth - after two weeks recovery period. Indicators of cellular immunity were determined by immunophenotyping and dyeing on Romanowsky-Giemsa. The level of immunoglbulins in blood serum was determined by radial immunodiffusion on Mancini. The level of cortisol in blood serum was determined-by immunoenzyme method. We determined that leukogram redistribution in favor of granulocyte fractions can be observed in the analyzed group in the intersession period, and also the reduction of the relative and absolute number of lymphocytes expressing antigens CD3, CD5, CD4 and CD 16. Also, all examined had reduced immunoregulatory index CD4(+)/CD8(+) and increased concentration of serum immunoglobulin G. Emotional stress increased earlier revealed tendencies. As a result the relative(23.41±1.01%) and absolute (0.28±0.02x10(9)/) number of T cells with the phenotype CD4(+), and immunoregulatory index (1.04±0.03) reached values lower than homeostatic norm. These indicators also show the longest period of recovery. Dynamics of the analyzed indices in session and recovery periods gives a reason to believe that mechanisms of adaptation and deadaptation are better displayed for thymus-dependent cells. Additional short-term stress factor intensifies immuno-suppression caused by living in radiation contaminated areas, and creates the basis for regular monitoring of health status of the population that suffered from the combined stress and for implementing preventive measures during the recovery period.

Relation of Neutrophil to Lymphocyte Ratio With Periprocedural Myocardial Damage in Patients Undergoing Elective Percutaneous Coronary Intervention

Neutrophil to lymphocyte ratio (NLR) has been proposed as a marker of cardiovascular risk. The potential relation between NLR and periprocedural myocardial damage after percutaneous coronary intervention (PCI) is unclear. We enrolled 502 consecutive patients with stable coronary artery disease undergoing elective PCI. Blood samples were drawn in all patients at baseline, 6 hours, and 24 hours after PCI for complete blood cell count and cardiac biomarkers (creatine kinase-MB and troponin T [Tn-T]) assessment. NLR was calculated as the ratio between the absolute number of neutrophil over the absolute number of lymphocyte. Periprocedural myocardial infarction (PMI) was defined according to the 2012 universal definition of myocardial infarction. In the overall population, a significant postprocedural increase in NLR was observed (3.255 [2.763 to 3.995] at baseline, 4.430 [3.390 to 6.020] at 6hours, 4.720 [3.940 to 5.750] at 24 hours, p <0.0001). PMI occurred in 33 patients (6.6%). Baseline NLR was similar in patients with and without PMI (3.250 [2.820 to 3.885] vs 3.260 [2.750 to 4.000], p= 0.898); however, patients who developed PMI showed significantly higher NLR both at 6 hours (5.750 [4.360 to 9.095] vs 4.370 [3.370 to 5.950], p <0.001) and 24hours (5.180 [4.440 to 8.065] vs 4.670 [3.920 to 5.710], p= 0.003). Among patients who developed PMI, periprocedural NLR increase showed a moderate positive correlation with both creatine kinase-MB (rho= 0.377, p= 0.031) and troponin T increase (rho= 0.506, p= 0.003). In conclusion, preprocedural NLR values do not impact on the occurrence of PMI during elective PCI; however, PCI procedures induce a significant increase in NLR that seems to be proportional to the magnitude of periprocedural myocardial damage.

Hematological and Growth Performance Studies after Withania Somnifera Supplementation in Broilers

This research was made to study the influence of addition of powdered root of Ashwaganda (Withania somnifera)on general health condition (growth performance) as well as hematological profile of broiler chicks. A 100 broiler chicks at first day after hatching were obtained from Wadi Company, Egypt, and were distributed at random into four equal groups. The first group received control ration only and kept as control group (Gr 1), the 2nd (Gr 2) ,3rd (Gr 3) and 4th (Gr 4) groups received control ration + 0.5%, 1% and 1.5% Withania somnifera root powder respectively for 42 days. Body weights, body weight gain, feed intake and feed efficiency were recorded on regular basis and hematological profile of the birds was estimated at the end of the 4th and 6th weeks of the research. Results showed that body weight and weight gain were significantly increased (P<0.05) in Gr2 and Gr3. While Gr4 showed non-significant changes in comparison with control group.Feed intake and feed efficiency were significantly decreased (P<0.05) in Gr2 and Gr3. While Gr4 showed non-significant changes in comparison with control group. Hematological results showed significant (P<0.05) rise in RBCs, PCV as well as Hb concentration in Gr2 and Gr3. While Gr4 showed non-significant changes in comparison with control group. Regarding toMCV (fl), MCH (pg) and MCHC (%)there were non-significant changes in all groups supplemented by Withania somnifera powdered root 0.5, 1 and 1.5% in comparison with control group. There were significant increase (P<0.05) in total leukocytic count and absolute numbers of lymphocytes in Gr2 and Gr3 over control group. These results indicate that addition of 0.5 and 1% Withania somnifera powdered root significantly enhanced general health condition (growth performance) as well as hematological profile of broiler chicks.

IFNL3/4 genotype is associated with altered immune cell populations in peripheral blood in chronic hepatitis C infection.

Single-nucleotide polymorphisms near the interferon lambda 3 (IFNL3) gene predict outcomes to infection and anti-viral treatment in hepatitis C virus (HCV) infection. To identify IFNL3 genotype effects on peripheral blood, we collected phenotype data on 400 patients with genotype 1 chronic hepatitis C (CHC). The IFNL3 responder genotype predicted significantly lower white blood cells (WBCs), as well as lower absolute numbers of monocytes, neutrophils and lymphocytes for both rs8099917 and rs12979860. We sought to define the WBC subsets driving this association using flow cytometry of 67 untreated CHC individuals. Genotype-associated differences were seen in the ratio of CD4CD45RO+ to CD4CD45RO− CD8CD45RO+ to CD8CD45RO−, NK CD56 dim to bright and monocyte numbers and percentages. Whole blood expression levels of IFNL3, IFNLR1 (interferon lambda receptor 1), IFNLR1-mem (a membrane-associated receptor), IFNLR1-sol (a truncated soluble receptor), MxA and T- and NK (natural killer) cell transcription factors TBX21, GATA3, RORC, FOXP3 and EOMES in two subjects were also determined. CHC patients demonstrated endogenous IFN activation with higher levels of MxA, IFNLR1, IFNLR1-mem and IFNLR1-sol, and IFNL3 genotype-associated differences in transcription factors. Taken together, these data provide evidence of an IFNL3 genotype association with differences in monocyte, T- and NK cell levels in the peripheral blood of patients with CHC. This could underpin genotype associations with spontaneous and treatment-induced HCV clearance and hepatic necroinflammation.

Commensal metabolite indol-3-propionic acid promotes gut barrier function by regulating IL-22 production during intestinal inflammatory conditions

Abstract Microbiota-derived metabolites provide signals to maintain homeostasis of the epithelial cell layer in the intestinal mucosa that in turn shape immune responses to bacterial pathogens. In contrast, disruption of these signals can result in dysregulated responses to microbiota that cause inflammatory bowel diseases (IBD). Here we tested the hypothesis that a metabolite produced via tryptophan metabolism by commensal bacteria, Indol-3-propionic acid (IPA), promotes immune homeostasis and gut barrier function in the intestinal mucosa during inflammatory conditions and prevents IBD. We investigated the effect of IPA on the intestinal mucosa using a model of colitis caused by oral infection with Citrobacter rodentium (Crod). Remarkably, we observed that IPA treatment results in protection of mice from colitis. Our studies show that IPA treatment results in increased expression of IL-22 by intraepithelial lymphocytes (IELs) of the small intestine (S). In fact, most of the IL-22 observed at 4 days following oral Crod infection is produced by IELs from small intestines. Since we have previously shown that IPA is a potential ligand of the nuclear receptor pregnane X receptor (PXR), which contributes to the maintenance of the intestinal barrier function, we further investigated if IPA protects by activating PXR or other pathways in the gut mucosa. Indeed we discovered an increase in the absolute numbers of IL-17A and IFNγ producing lymphocytes from the IEL compartment of the colon by the IPA treatment and this effect was dependent on PXR expression. In conclusion, we believe the discovery of metabolites like IPA that promotes the repair of the intestinal epithelial cell layer can provide new strategies to prevent and treat IBD.

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100 children were observed. Tuberculosis of inrathoracic lymph nodes was found in 79 (79,0 %) patients, complicated tuberculosis � in 29 (36,7 %) of 79 patients. The phase of reverse development of tuberculosis was detected in 47,0 % of children with TB. Due to the slow dynamics of the disease, the duration of chemotherapy in 75,0 % of children was 240 days. In the mode of treatment, drug resistance of a patient in lesion of tuberculosis infection is taken into account. As a result of treatment, resorption of infiltration in the lungs was observed in 15,0 %, fibrosis � in 26,0 %, formation of small calcifications � in 39,0 %, formation of multiple large calcifications � in 20,0 % of children. The cause of the formation of calcifications is the late detection of tuberculosis. In children with the formation of calcifications throughout the course of treatment the number of leukocytes decreased, the downward trend in the absolute number of lymphocytes to the 4th month of treatment was traced, the content of leukocytes increased to the end of primary treatment. In children without formation of calcifications the reduction of leukocytes and lymphocytes was not identified during the treatment. The decrease in the content of leukocytes and lymphocytes may explain long-term persistence of endogenous intoxication.

Lymphocyte subpopulations in premature infants: an observational study

Background and objectivesThe infant's immune system evolves over the first months and years of life. Strong correlation exists between lymphocyte count, lymphocyte subpopulations and gestational age at birth. Associations with...

CYTOTOXIC LYMPHOCYTES: THE ROLE IN INFLAMMATION IN PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE EXACERBATION AND REMISSION

Background: Despite the number of publications related to the expression of surface antigens of periphery blood lymphocytes in chronic obstructive pulmonary disease (COPD), algorithm for interpreting of the results and implicating pathogenetic treatments still needs to be developed. Aim: To assess the role of cytotoxic lymphocytes in the maintaining of inflammation in COPD. Materials and methods: To examine immune status in 37 patients with COPD exacerbation or remission and 24 healthy donors (control group), blood cytotoxic T-lymphocytes and NK-cells contents were measured using indirect immunofluorescence method. Absolute and relative numbers of lymphocytes expressing CD3, CD4, CD8, CD16, CD20, CD23, CD25, CD54, CD71, CD72, HLA-DR, CD95 antigens, membrane immunoglobulins M (mIgM) and G (mIgG) were estimated. Results: In COPD., significantly increased numbers of blood cytotoxic lymphocytes were demonstrated independently from the disease stage (p < 0.001). During COPD exacerbation, significant elevations of CD4, CD8, CD20, CD72, NK-cells numbers, serum mIgM and mIgG were demonstrated. During remission, CD20 and CD72 content returned to normal, though, increased numbers of other cytotoxic cells persisted promoting inflammation and progressive damage of pulmonary and bronchial tissues. Conclusion: Observed changes may be due to excessive stimulation of T-cell component of immune system in COPD patients both in exacerbation and remission. Relative reduction of total T-lymphocyte numbers indicates non-specific (non-infectious) inflammation type. High cytotoxic potential of immune system results in pulmonary damage and promotes development of pneumosclerosis and emphysema.

The clinical and immunological indices and their interaction with thyroid status in patients with Graves’ disease depending on thyrocytes peroxidase autoantibodies level

The process of antibody production against thyroid peroxidase and their relation to the complement-mediated cytotoxicity have the most pronounced cytotoxic effect on thyrocytes in Graves’ disease.Aim.To study the clinical and immunological indices and their interaction with thyroid status in patients with Graves’ disease depending on thyrocytes peroxidase autoantibodies level.Material and methods.The study included 35 women aged 18 to 55 years, mean age of 39.1±7.2, with verified diagnosis of Graves’ disease for the first time, before antithyroid therapy. The clinical and immunological parameters and their relationship with thyroid status studied depending on the level of AT-TPO. The definition of thyroid hormones content was measured using immunoradiometricassay analysis with standard test kits. AT-TPO was assessed by ELISA. Population and subpopulation composition of blood lymphocytes was evaluated using the method of indirect immunofluorescence using monoclonal antibodies to CD3, CD4, CD8, CD16, CD19 and HLA-DR. The humoral subset pattern characterized by the relative synthesis of IgA/CD19+, Ig M/CD19+, IgG/CD19+.Results.The most significant changes in population and subpopulation composition of lymphocytes was found in patients with more than 100 IU/L AT-TPO level and characterized by an increase in absolute lymphocyte, relative and absolute number of CD19+ and HLA-DR+cells, and CD3+ and CD8+cells, as compared to the control range and the values identified in group with AT-TPO less than 100 IU/L. A strong negative relationship AT-TPO with the number of T-lymphocytes (r=–0.75; p&lt;0.001) and moderate with the relative synthesis level of IgM (r=–0,53; p=0.017) have been revealed. In Graves’ disease patients with AT-TPO more than 100 IU/l levels the statistically significant relationships were not found.Conclusion.The immunopathogenesis of Graves’ disease is characterized by positive correlation in AT-TPO and indicators of b-cell immunity, and negative — with the parameters of T-cell immunity, regardless of thyrocytes peroxidase autoantibodies concentration.

Serological and molecular detection of bovine leukemia virus in cattle in Iraq

Bovine leukemia virus (BLV) is highly endemic in many countries, including Iraq, and it impacts the beef and dairy industries. The current study sought to determine the percentage of BLV infection and persistent lymphocytosis (PL) in cattle in central Iraq. Hematological, serological, and molecular observations in cross breeds and local breeds of Iraqi cattle naturally infected with BLV were conducted in the peripheral blood mononuclear cells of 400 cattle (340 cross breed and 60 local breed) using enzyme-linked immunosorbent assay and polymerase chain reaction (PCR). On the basis of the absolute number of lymphocytes, five of the 31 positive PCR cases had PL. Among these leukemic cattle, one case exhibited overt neutrophilia. Serum samples were used to detect BLV antibodies, which were observed in 28 (7%) samples. PCR detected BLV provirus in 31 samples (7.75%). All 28 of the seropositive samples and the 3 seronegative samples were positive using PCR. Associations were observed between bovine leukosis and cattle breed, age and sex. Age-specific analysis showed that the BLV percentage increased with age in both breeds. Female cattle (29 animals; 7.34%) exhibited significantly higher infectivity than male cattle (two animals; 4.34%). In conclusion, comprehensive screening for all affected animals is needed in Iraq; programs that segregate cattle can be an effective and important method to control and/or eliminate the BLV.Emerging Microbes and Infections (2016) 5, e56; doi:10.1038/emi.2016.60; published online 8 June 2016