Phenotype Of Blood Lymphocytes Research Articles

Precision medicine using molecular-target drugs in psoriatic arthritis.

Psoriatic arthritis (PsA) presents various clinical manifestations, including skin lesions, peripheral arthritis, axial involvement, enthesitis, nail involvement, dactylitis, and uveitis. In addition, it causes a high incidence of lifestyle-related diseases and an increase in cerebrovascular and cardiovascular events. As the pathology of PsA has been clarified, molecular-targeted drugs targeting tumor necrosis factor-α, interleukin (IL)-17A, IL-17A/F, IL-17 receptor, IL-12/23(p40), IL-23p19, Cytotoxic T-lymphocyte Antigen-4 (CTLA-4), Janus kinase, and phosphodiesterase-4 have been developed and are widely used in clinical practice. PsA is clinically and molecularly heterogeneous, and it is necessary to improve various clinical symptoms with limited treatment options simultaneously; therefore, rheumatologists sometimes encounter difficult situations in clinical practice. Hence, the development of precision medicine may improve treatment outcomes. Recently, the strategic use of molecular-targeted drugs based on the stratification of patients with PsA by peripheral blood lymphocyte phenotyping and serum cytokine concentrations has been reported to possibly lead to a higher therapeutic response. A randomized controlled trial was initiated to verify the efficacy of this treatment strategy. However, to make precision medicine in PsA feasible, shifting from conventional clinical trials to clinical trials based on biomarker profiles and accumulating further data are necessary.

Characterizing Human Peripheral Blood Lymphocyte Phenotypes and Their Correlations with Body Composition in Normal-Weight, Overweight, and Obese Healthy Young Adults.

Background and Objectives: Obesity-associated chronic low-grade inflammation supports various systemic alterations. In this descriptive study, 122 apparently healthy adults aged 20 to 35 years were voluntarily included and classified based on body mass index (BMI) as normal-weight (NW), overweight (OW), and obese (OB). This study aims to characterize peripheral blood (PB) lymphocyte (Ly) phenotypes and investigate their correlations with body composition indices (BCIs) in healthy young adults. Materials and Methods: The following BCIs were measured: waist circumference, hip circumference, height, waist-to-hip ratio, waist-to-height ratio, total body fat mass, visceral fat level, weight, and BMI. White blood cell count (WBC), Ly absolute count, serum TNF-α, and IFN-γ were quantified. Ly subpopulations were analyzed as follows: total TLy (TTLy-CD45+CD3+), early activated TLy (EATLy-CD45+3+69+), total NKLy (TNKLy-CD45+CD3-CD56+CD16+), NKdim (low expression of CD56+), NKbright (high expression of CD56+), BLy (CD45+CD3-CD19+), T helper Ly (ThLy-CD45+CD3+CD4+), and T cytotoxic Ly (TcLy-CD45+CD3+CD8+). Results: Higher BMI has significantly higher WBC and BLy (p < 0.0001; p = 0.0085). EATLy significantly decreased from NW to OB (3.10-NW, 1.10-OW, 0.85-OB, p < 0.0001). Only EATLy exhibited significant negative correlations with all the BCIs. A significantly higher TNF-α was observed in the OW and OB groups compared to the NW group. IFN-γ increased linearly but nonsignificantly with BMI. TTLy showed a nonsignificant positive correlation with both IFN-γ and TNF-α, while EATLy showed a negative correlation, significant only for IFN-γ. NKLy subpopulations exhibited a consistent negative correlation with TNF-α, significant only for NKdim (p = 0.0423), and a nonsignificant consistent positive correlation with IFN-γ. A nonsignificant negative correlation between age and both TNKLy (r = -0.0927) and NKdim (r = -0.0893) cells was found, while a positive correlation was found with NKbright (r = 0.0583). Conclusions: In conclusion, the baseline immunological profile of PB is influenced by excessive adipose tissue in healthy young adults.

THE CO-ADMINISTRATION OF POIL-2 WITH TGEV INACTIVATED VACCINE ENHANCES IMMUNE RESPONSE OF PIGLETS TO TGEV

Porcine interleukin-2 (poIL-2) has not yet been demonstrated to be immune-enhancing against porcine transmissible gastroenteritis virus (TGEV) inactivated vaccine (IV), despite IL-2 having been proven to have immunological adjuvant effects for a variety of vaccinations. In this work, the impact of poIL-2 on TGEV IV in terms of immunological enhancement was investigated. Twenty four (24) SPF piglets were utilized and divided into six groups: PBS group, poIL-2 group, IV group, 10μg poIL-2+IV group, 50μg poIL-2 +IV group, and 250μg poIL-2+IV group. They received a second vaccine at 28 days point following the initial immunization. Serum and blood samples were obtained at various periods throughout the experiment. By using ELISA assay, neutralization assay, MTT assay, and flow cytometry assay, the TGEV-specific antibody expressions, neutralizing antibodies generations, interleukin-4 (IL-4), and interferon-gamma (IFN-γ) productions, peripheral blood mononuclear cells (PBMCs) proliferation response and lymphocyte phenotype subpopulations (CD3+, CD4+, and CD8+ immune cells) reflections were determined. The results showed that piglets inoculated with IV supplemented with poIL-2 significantly not only increased more piglet cellular immunity against TGEV by raising the degrees of IL-4, IFN-γ, Stimulation Index (SI), and the ratio of CD4+ /CD8+ cell subgroups, but also promoted more humoral immunity against TGEV by increasing levels of anti-TGEV specific antibodies and neutralizing antibodies (NAs) than those piglets inoculated with the TGEV IV alone. Additionally, the results suggested that porcine interleukin-2 (poIL-2) may improve pigs' immune responses in a dosage-dependent way.Our study revealed that poIL-2 had an immune-enhancing effect on the immunization of TGEV IV, and it possessed the potential to be applied as an immune-stimulating agent. Keywords: Inactivated vaccine (IV); adjuvant; porcine transmissible gastroenteritis virus (TGEV); porcine interleukin-2 (poIL-2); immune response.

Addition of CD14 improves discrimination of lymphocytes in the TBNK phenotyping panel.

Peripheral blood lymphocyte phenotyping panels typically include CD45 for discrimination of the lymphocyte population, and fluorophore-conjugated monoclonal antibodies to identify T, B, and Natural Killer (NK) cells. While CD45 combined with side scatter is generally sufficient to clearly distinguish lymphocytes from monocytes in the majority of peripheral blood samples, it is challenging to accurately gate lymphocytes in samples from patients with monocytosis or significant lymphopenia, or from very young infants. Addition of a monocyte marker to lymphocyte phenotyping panels for monocyte exclusion has previously been evaluated for improved discrimination of lymphocytes, albeit largely in healthy donor adult samples. Here we evaluate the effect of the addition of CD14 to a standard lymphocyte phenotyping panel on total lymphocyte, T, B, and NK cell percentages in a predominantly pediatric population of patients under evaluation chiefly for immunodeficiency, immune-depletion, or immune reconstitution. Addition of CD14 to the standard lymphocyte phenotyping improved discrimination of lymphocytes from monocytes, resulted in decreased NK cell percentages, likely because CD16+ and/or CD56+ monocytes were included in the CD56+CD16+ NK cell gate with conventional gating, and although less significant, resulted in an increased percentage of B cells, since relatively larger B cells were likely gated out by more restrictive light scatter gating used with the conventional gating approach. The change in NK and B cell percentages were more pronounced in samples from patients below a year of age, and in patients who were relatively lymphopenic. These data suggest that addition of CD14 to conventional lymphocyte phenotyping panels that utilize CD45 versus side scatter gating results in significant improvement in the accuracy of lymphocyte gating, and accurate quantification of NK and B cells particularly in samples from infants and lymphopenic individuals.

Intracranial injection of natural killer cells engineered with a HER2-targeted chimeric antigen receptor in patients with recurrent glioblastoma.

Glioblastoma (GB) is incurable at present without established treatment options for recurrent disease. In this phase I first-in-human clinical trial we investigated safety and feasibility of adoptive transfer of clonal chimeric antigen receptor (CAR)-NK cells (NK-92/5.28.z) targeting HER2, which is expressed at elevated levels by a subset of glioblastomas. Nine patients with recurrent HER2-positive GB were treated with single doses of 1 × 107, 3 × 107, or 1 × 108 irradiated CAR-NK cells injected into the margins of the surgical cavity during relapse surgery. Imaging at baseline and follow-up, peripheral blood lymphocyte phenotyping and analyses of the immune architecture by multiplex immunohistochemistry and spatial digital profiling were performed. There were no dose-limiting toxicities, and none of the patients developed a cytokine release syndrome or immune effector cell-associated neurotoxicity syndrome. Five patients showed stable disease after relapse surgery and CAR-NK injection that lasted 7 to 37 weeks. Four patients had progressive disease. Pseudoprogression was found at injection sites in 2 patients, suggestive of a treatment-induced immune response. For all patients, median progression-free survival was 7 weeks, and median overall survival was 31 weeks. Furthermore, the level of CD8+ T-cell infiltration in recurrent tumor tissue prior to CAR-NK cell injection positively correlated with time to progression. Intracranial injection of HER2-targeted CAR-NK cells is feasible and safe in patients with recurrent GB. 1 × 108 NK-92/5.28.z cells was determined as the maximum feasible dose for a subsequent expansion cohort with repetitive local injections of CAR-NK cells.

Kidney allograft rejection is associated with an imbalance of B cells, regulatory T cells and differentiated CD28-CD8+ T cells: analysis of a cohort of 1095 graft biopsies.

The human immune system contains cells with either effector/memory or regulatory functions. Besides the well-established CD4+CD25hiCD127lo regulatory T cells (Tregs), we and others have shown that B cells can also have regulatory functions since their frequency and number are increased in kidney graft tolerance and B cell depletion as induction therapy may lead to acute rejection. On the other hand, we have shown that CD28-CD8+ T cells represent a subpopulation with potent effector/memory functions. In the current study, we tested the hypothesis that kidney allograft rejection may be linked to an imbalance of effector/memory and regulatory immune cells. Based on a large cohort of more than 1000 kidney graft biopsies with concomitant peripheral blood lymphocyte phenotyping, we investigated the association between kidney graft rejection and the percentage and absolute number of circulating B cells, Tregs, as well as the ratio of B cells to CD28-CD8+ T cells and the ratio of CD28-CD8+ T cells to Tregs. Kidney graft biopsies were interpreted according to the Banff classification and divided into 5 biopsies groups: 1) normal/subnormal, 2) interstitial fibrosis and tubular atrophy grade 2/3 (IFTA), 3) antibody-mediated rejection (ABMR), 4) T cell mediated-rejection (TCMR), and 5) borderline rejection. We compared group 1 with the other groups as well as with a combined group 3, 4, and 5 (rejection of all types) using multivariable linear mixed models. We found that compared to normal/subnormal biopsies, rejection of all types was marginally associated with a decrease in the percentage of circulating B cells (p=0.06) and significantly associated with an increase in the ratio of CD28-CD8+ T cells to Tregs (p=0.01). Moreover, ABMR, TCMR (p=0.007), and rejection of all types (p=0.0003) were significantly associated with a decrease in the ratio of B cells to CD28-CD8+ T cells compared to normal/subnormal biopsies. Taken together, our results show that kidney allograft rejection is associated with an imbalance between immune cells with effector/memory functions and those with regulatory properties.

A219 BIOLOGIC USE DURING PREGNANCY IN WOMEN WITH INFLAMMATORY BOWEL DISEASE AND ASSOCIATED NEONATAL OUTCOMES: A RETROSPECTIVE CHART REVIEW

Abstract Background Biologics are the mainstay of therapy for patients with advanced inflammatory bowel disease (IBD). Most biologics readily undergo placental transfer during the second trimester and remain detectable in infant serum for up to 12 months. Therefore, most clinical guidelines currently recommend avoiding live vaccines in the first 6 to 12 months for exposed infants. In Canada, the rotavirus vaccine is the only live vaccine administered before 6 months. Emerging evidence is suggesting that the vaccine may safely be given to infants with normal immune function, even if serum biologic level is detectable. Specialist assessment of infant immune function may help guide decision-making about rotavirus vaccine administration, but there remains a large gap in our knowledge of the impact of biologic exposure on the immune function of infants, necessitating further study. Purpose In this single-centre retrospective chart review, we aim to study the clinical outcomes, immune function, and rotavirus vaccine recommendations for infants exposed to biologics in utero during their first year of life. Method The study included mothers seen at the Pregnancy IBD Clinic at Mount Sinai Hospital in Toronto, ON, who were offered a referral to the Special Immunization Clinic (SIC) at The Hospital for Sick Children due to biologic exposure in utero. Data was collected on the recommendations made by paediatric specialists at SIC, based on complete blood count, lymphocyte phenotyping, and T-cell receptor excision analysis. Data was obtained on adverse neonatal outcomes in the first year of life, including prematurity, congenital malformations, and infections based on post-partum surveys completed by mothers and follow-up letters from SIC. Result(s) 43 patients were referred to and seen by paediatric specialists at SIC between 2 to 12 months of age. 18 infants (42%) were exposed to Adalimumab in utero, 16 (37%) to Infliximab, 5 (12%) to Vedolizumab, and 4 (9%) to Ustekinumab. The rotavirus vaccine was recommended to 34 infants (81%) and not recommended to 3 (7%) for reasons including gastrointestinal illness, neutropenia, and low lymphocyte counts. Recommendation is pending for 6 infants (14%). Two infants (5%) had premature births. Four infants (9%) were admitted to the NICU for reasons including respiratory distress, and prematurity (9%). One infant (2%) had a congenital malformation, specifically bilateral sensorineural hearing loss. Six infants (14%) had upper respiratory tract infections, none of which required hospital admission or antibiotics. Conclusion(s) This study is currently in progress. Further data is required to assess whether biologic exposure in utero has a significant impact on neonatal immune function, especially beyond the first year of life, and whether there is a significant difference in vaccine recommendations and response based on the type of biologic received. Disclosure of Interest None Declared

Transient hypogammaglobulinaemia of the young in a Thoroughbred yearling

SummaryA 12‐month‐old Thoroughbred colt was presented for chronic recurrent respiratory disease characterised by persistent nasal discharge, intermittent pyrexia and coughing of several months' duration. These episodes were responsive to empirical antimicrobial therapy but returned once therapy was discontinued. History, physical examination and initial diagnostics were suggestive of bacterial pneumonia and possible underlying immunodeficiency. Serum immunoglobulin analysis and peripheral blood lymphocyte phenotyping revealed low serum IgG and IgM concentrations. In this 12‐month‐old colt, low serum IgG concentration was the most diagnostic parameter and supported a presumptive diagnosis of transient hypogammaglobulinaemia of the young. The colt responded favourably to repeated treatment with antimicrobials for bacterial pneumonia, and serial immunologic testing over the following months showed gradual increase in serum IgG and IgM concentrations, supporting the diagnosis of delayed development of humoral immunity in this patient.

Phenotype of peripheral blood lymphocytes in angina

The phenotype of peripheric blood lymphocytes is studied in 87 patients with angina by cytofluorimetry. It is shown that the course of the disease is accompanied by significant changes of the phenotype of lymphocytes: the increase of expression of activation markers, the increase of the content of CD16+cells. However, the changes of the phenotype of lymphocytes in groups of patients with lacunar and ulceromembranous anginas are ambiguous in the acute period of the disease as well as late in the observation.

FEATURES OF TIM-3, CD9, CD49a MOLECULE EXPRESSIONS BY PERIPHERAL BLOOD LYMPHOCYTES DURING PHYSIOLOGICAL PREGNANCY

Physiological pregnancy is characterized by significant changes in the phenotype and functions of peripheral blood lymphocytes. Thus, the number of lymphocytes migrating from peripheral blood in the uterus and expressing Tim-3, CD49a, CD9 molecules which limit their cytotoxicity and participate in their migration, is significantly increased in the first half of pregnancy. However, the expression of these molecules on peripheral blood lymphocytes during physiological pregnancy has not been studied. The aim of the work was to study the expression of Tim-3, CD49a, CD9 molecules in the total pool of lymphocytes and on CD3⁺T-lymphocytes of peripheral blood of women in the I and III trimesters of physiological pregnancy. The object of the study was the peripheral blood of healthy women with physiological pregnancy in the I and III trimesters. The comparison group is healthy non-pregnant women in the first phase of the menstrual cycle. The expression of Tim3, CD49a, and CD9 molecules was analyzed in a pool of lymphocytes and on CD3⁺T-lymphocytes of peripheral blood by flow cytometry. It was found that in the first trimester, the expression of Tim-3 and CD49a in the total pool of lymphocytes was higher, and CD9 did not change compared to non-pregnant women. In the third trimester, the expression of Tim-3 and CD49a in the total pool of lymphocytes did not change, and CD9 was lower than in non-pregnant women. The expression of Tim-3 did not change on CD3⁺-T lymphocytes in both I and III trimesters, and CD49a and CD9 were lower compared to non-pregnant women. Thus, the expression of Tim-3, CD49a, and CD9 molecules varied during the trimesters of pregnancy both in the total pool of lymphocytes and CD3⁺-T lymphocytes of peripheral blood, which is significant for the regulation of their cytotoxic function and migration activity.

Bowel Histology of CVID Patients Reveals Distinct Patterns of Mucosal Inflammation

Diarrhea is the commonest gastrointestinal symptom in patients with common variable immunodeficiency (CVID). Different pathologies in patients’ bowel biopsies have been described and links with infections have been demonstrated. The aim of this study was to analyze the bowel histology of CVID patients in the Royal-Free-Hospital (RFH) London CVID cohort. Ninety-five bowel histology samples from 44 adult CVID patients were reviewed and grouped by histological patterns. Reasons for endoscopy and possible causative infections were recorded. Lymphocyte phenotyping results were compared between patients with different histological features. There was no distinctive feature that occurred in most diarrhea patients. Out of 44 patients (95 biopsies), 38 lacked plasma cells. In 14 of 21 patients with nodular lymphoid hyperplasia (NLH), this was the only visible pathology. In two patients, an infection with Giardia lamblia was associated with NLH. An IBD-like picture was seen in two patients. A coeliac-like picture was found in six patients, four of these had norovirus. NLH as well as inflammation often occurred as single features. There was no difference in blood lymphocyte phenotyping results comparing groups of histological features. We suggest that bowel histology in CVID patients with abdominal symptoms falls into three major histological patterns: (i) a coeliac-like histology, (ii) IBD-like changes, and (iii) NLH. Most patients, but remarkably not all, lacked plasma cells. CVID patients with diarrhea may have an altered bowel histology due to poorly understood and likely diverse immune-mediated mechanisms, occasionally driven by infections.

Preoperative administration of corticosteroids in thoracic anesthesiology

Annotation. One lung ventilation (OLV) is one of the most difficult intraoperative methods of respiratory support for anesthesiologists. OLV should provide the most comfortable surgical field, maintaining proper gas exchange and minimizing damage to both lungs. This anesthetic procedure has a significant inflammatory response, so using perioperative corticosteroid therapy to suppress inflammatory mediators is recommended as an approach to improving prognosis. Therefore, the aim of this study was to determine how the preoperative administration of methylprednisolone affects the systemic proinflammatory response of cytokines during thoracic surgery. The analysis was performed for 80 patients who underwent surgeries in the thoraco-abdominal department of the Shalimov National Institute of Surgery and Transplantology. Patients were divided into 2 groups (study – 40 patients who were administered methylprednisolone 10 mg / kg intravenously during induction of anesthesia and control – 40 patients without methylprednisolone). Before surgery and in the postoperative period on days 1, 3 and 5, the surface phenotype of peripheral blood lymphocytes and the expression of IL-6 by monocytes were determined by flow cytofluorometry. The author's MedStat package was used for statistical analysis (Lyakh Yu.E., Guryanov V.G., 2004–2012). Postoperative indicators of IL-6 monocyte expression in the blood of patients administered methylprednisolone were significantly lower on the 1st and 3rd postoperative day (p&lt;0.001). Thus, preoperative administration of methylprednisolone reduces the release of pro-inflammatory cytokines and improves the condition of patients after thoracic surgery. We consider it expedient to conduct further research on the administration of methylprednisolone for several days in the postoperative period.

Peripheral blood lymphocyte phenotypes in Alzheimer and Parkinson’s diseases

IntroductionNeuroinflammation is involved in the pathophysiology of various neurological disorders, in particular Alzheimer disease (AD) and Parkinson’s disease (PD). Alterations in the blood-brain barrier may allow peripheral blood lymphocytes to enter the central nervous system; these may participate in disease pathogenesis. ObjectiveTo evaluate the peripheral blood lymphocyte profiles of patients with AD and PD and their association with the disease and its progression. MethodsThe study included 20 patients with AD, 20 with PD, and a group of healthy individuals. Ten of the patients with AD and 12 of those with PD were evaluated a second time 17 to 27 months after the start of the study. Lymphocyte subpopulations and their activation status were determined by flow cytometry. All patients underwent neurological examinations using internationally validated scales. ResultsCompared to healthy individuals, patients with AD and PD showed significantly higher levels of activated lymphocytes, lymphocytes susceptible to apoptosis, central memory T cells, and regulatory T and B cells. As the diseases progressed, there was a significant decrease in activated cells (CD4+ CD38+ and CD8+ CD38+ in PD and AD, CD4+ CD69+ and CD8+ CD69+ in PD), T cells susceptible to apoptosis, and some regulatory populations (CD19+ CD5+ IL10+ in PD and AD, CD19+ CD5+ IL10+ FoxP3+, CD4+ FoxP3+ CD25+ CD45RO+ in PD). In patients with AD, disease progression was associated with lower percentages of CD4+ CD38+ cells and higher percentages of effector CD4 cells at the beginning of the study. Significant differences were observed between both diseases. ConclusionsThis study provides evidence of changes in peripheral blood lymphocyte phenotypes associated with AD and PD and their severity. Considering effective blood-brain communication, our results open new avenues of research into immunomodulation therapies to treat these diseases.

Phenotypic profile of peripheral blood lymphocytes in patients with inflammatory bowel diseases

Mechanisms of recognition and effector responses of immune system upon initiation and maintenance of immune-mediated inflammation and tissue damage in inflammatory bowel diseases, including Crohn’s disease and ulcerative colitis (UC), have been actively studied over recent time. Existing evidence suggests these diseases to be caused by abnormal immune response against intestinal flora microorganisms in genetically susceptible individuals. Despite available data on the features of immune disorders in ulcerative colitis and Crohn’s disease, there are still many questions about the involved minor lymphocyte subpopulations and contribution of various functionally active molecules, which play a key role in recognition and initiation of the immune response and can be considered biomarkers of a pathological process in inflammatory bowel diseases. The populations of T lymphocytes with γδT cell receptor, B1 cells and NK T lymphocytes are of greatest interest, as well as functionally active TLRs (Toll-like receptors), CD89, CD314, etc. Due to substantial progress in studying the nature of recognition and activation of the immune cells, the paper presents phenotypic and functional characteristics of major and minor subpopulations of peripheral blood lymphocytes observed in 25 patients treated at the Surgery Department of the State Institution “Minsk Regional Clinical Hospital” (Republic of Belarus) from 2018 to 2020. The detected changes in peripheral blood lymphocyte phenotype of inflammatory bowel diseases patients suggest distinct immunological profiles prevailing in the damage mechanisms in Crohn’s disease and ulcerative colitis. I.e., in Crohn’s disease patients, B1 lymphocytes with CD19+CD5+ and NK cells in combination with increased CD56bright population, as well as NK T cells with anti-inflammatory and regulatory activity are involved into genesis of the disease. In ulcerative colitis, T, B, NK lymphocytes with pro-inflammatory phenotype and T lymphocytes with γδ T cell receptor may play a pathogenetic role in maintenance of chronic inflammation. With respect to functional significance of activating receptors, the number of TLR4- и CD89-positive cells may be used for developing immunological criteria/ biomarkers of therapeutic efficacy of new drugs. Studying interactions between innate and adaptive immunity will open new perspectives in understanding immunological disorders associated with chronic gastrointestinal inflammation.

SUBPOPULATION STRUCTURE OF PERIPHERAL BLOOD LYMPHOCYTES OF DONORS

Introduction.Long-term monitoring of immune system parameters in cancer patient in FSBI “N.N. Blokhin Medical Research Center of Oncology” of the Ministry of Health of Russia, as well as the absence of an immunological research algorithm and a spectrum of significant markers, served as the basis for this study.Purpose.To present reference values and determine the normal range for subpopulations of systemic immunity lymphocytes.Materials and methods.The phenotype of peripheral blood lymphocytes was studied in 186 healthy donors (86 men and 100 women), mean age 41,9 ± 12,5 years. To assess the multivariate phenotype by flow cytometry, monoclonal antibodies to CD45, CD3, CD4, CD8, CD16, CD56, CD19, CD25, CD28, CD11b, CD127, HLA-Dr, TCR-γ / δ, Perforin and Granzime B labeled various fluorochromes.Results.A panel of markers of immunocompetent cells of systemic immunity was developed as a basis for assessing the state of the immune system of cancer patients. Reference values and normal boundaries are given for characterizing the linearity of cells with detailing of the phenotypic and functional heterogeneity of the population of CD8+-lymphocytes, activation markers and the pool of naive cells; characteristics of various types of regulatory cells, functional activity of cells of the effector link of immunity. A comparative analysis of immunoregulatory indices was carried out and the vulnerability of the formula CD3+CD4+/CD3+CD8+to the CD4+/CD8+index was proved.Conclusion.The spectrum of the proposed indicators is the product of many years of research carried out in the laboratory of clinical immunology of the Federal State Budgetary Institution “N.N. Blokhin National Medical Research Center of Oncology” оf the Ministry of Health of the Russian Federation, in order to search for significant criteria for assessing the state of the immune system in cancer.

Evaluation of an extruded diet for adult dogs containing larvae meal from the Black soldier fly (Hermetia illucens)

Insects have experienced an increasing interest as a protein source in recent years. However, their suitability as a protein source for dogs was not yet widely investigated. Therefore, the aim of the study was to compare two extruded diets for adult dogs containing either larvae meal from black soldier fly (Hermetia illucens; HI) or lamb meal as the predominant protein source with regard to apparent fecal nutrient digestibility and the influence on fecal and immunological parameters. Twelve Beagles were used in this cross-over study and divided into two groups. The test diet with 200 g/kg HI larvae meal (HI) was compared with a control diet containing lamb meal (CON). Dogs were fed each diet for a period of five weeks. Blood samples for a complete blood count, lymphocyte phenotyping and lymphocyte proliferation tests were taken at the end of each feeding period. The latter was either mitogen- or feed antigen-induced with different protein concentrations of HI larvae meal (30–500 μg protein/mL). Fecal samples were collected for five days during each feeding period. The coefficient of the total apparent fecal digestibility (CTTAD) of macronutrients, fecal metabolites, the output and the dry matter content of the feces were analyzed. d-Glucosamine as indicator for chitin was analyzed by high-performance anion-exchange chromatography with pulsed amperometric detection in the diet and the feces. The chitinase activity was determined in the feces. The fecal output was reduced in dogs fed the HI diet (P = 0.038). The CTTAD of dry matter was higher (P = 0.002) when dogs were fed HI compared to CON (0.832 vs. 0.818) and the CTTAD of crude protein was higher (P = 0.002) when dogs received CON compared to HI (0.792 vs. 0.773). Concentrations of acetate (P = 0.011) and ammonium (P = 0.001) were higher in CON. The fecal score differed (P = 0.041) between HI (2.25) and CON (2.04), but both feeding groups had a formed and dry fecal consistency. The chitin concentration was numerically higher in the feces of dogs fed HI (92.2 g/kg DM) compared to CON (2.9 g/kg DM), the fecal chitinase activity did not differ between both diets. Complete blood counts, lymphocyte phenotyping and proliferation were not affected. In conclusion, the HI larvae meal in a dry dog diet was tolerated without adverse signs and did not affect immunological measurements compared to a commercial dry diet with lamb meal, indicating that larvae meal from HI can be considered an “alternative” protein source for dog nutrition.

Peripheral Blood Lymphocyte Phenotype Differentiates Secondary Antibody Deficiency in Rheumatic Disease from Primary Antibody Deficiency.

The phenotype of primary immunodeficiency disorders (PID), and especially common variable immunodeficiency (CVID), may be dominated by symptoms of autoimmune disorders. Furthermore, autoimmunity may be the first manifestation of PID, frequently preceding infections and the diagnosis of hypogammaglobulinemia, which occurs later on. In this case, distinguishing PID from hypogammaglobulinemia secondary to anti-inflammatory treatment of autoimmunity may become challenging. The aim of this study was to evaluate the diagnostic accuracy of peripheral blood lymphocyte phenotyping in resolving the diagnostic dilemma between primary and secondary hypogammaglobulinemia. Comparison of B and T cell subsets from patients with PID and patients with rheumatic disease, who developed hypogammaglobulinemia as a consequence of anti-inflammatory regimes, revealed significant differences in proportion of naïve B cells, class-switched memory B cells and CD21low B cells among B cells as well as in CD4+ memory T cells and CD4+ T follicular cells among CD4+ T cells. Identified differences in B cell and T cell subsets, and especially in the proportion of class-switched memory B cells and CD4+ T follicular cells, display a considerable diagnostic efficacy in distinguishing PID from secondary hypogammaglobulinemia due to anti-inflammatory regimens for rheumatic disease.

Transportation and Routine Veterinary Interventions Alter Immune Function in the Dog

Rapid activation of the hypothalamic pituitary adrenal axis and the sympathetic nervous system are hallmarks of the acute stress response and these systems interact with the immune system by signaling though glucocorticoid and adrenergic receptors on immune cells. There is limited information about the effect of these physiologic responses on immunologic parameters of pet dogs enrolled in clinical studies. The objective of this study was to evaluate how travel, instrumentation, and hospitalization alter immunologic parameters in pet dogs. Blood was collected from healthy dogs in a home environment and from healthy dogs at the time of presentation to the hospital and after instrumentation and 24 hours of hospitalization. We found that lipopolysaccharide (LPS)-induced downregulation of toll like receptor 4 (TLR4) was blunted in dogs exposed to stress. Neutrophil and monocyte major histocompatibility complex class II (MHCII) expression increased after transportation to the veterinary hospital but then became similar to that of the control dogs at the end of hospitalization. Peripheral blood mononuclear cell cytotoxicity function was blunted in dogs exposed to the stress of transportation as well as hospitalization. Neutrophil apoptosis was greater in dogs exposed to stress compared to controls although this effect significantly decreased after hospitalization stress. Conversely, stress did not alter induced or spontaneous cytokine production from leukocytes, neutrophil or monocyte expression of TLR4, LPS-induced downregulation of monocyte TLR4, LPS-induced neutrophil and monocyte expression of MHCII or peripheral blood lymphocyte phenotype. Transportation and instrumentation/hospitalization stress should be considered when interpreting immunologic studies in pet dogs.

Peripheral blood lymphocyte subpopulations in patients with bipolar disorder type II

We investigated the phenotype of peripheral blood lymphocytes of patients with bipolar disorder type II in different phases of the disease in order to check whether there are specific changes in the immune parameters. Lymphocytes subpopulations were analyzed ex vivo with flow cytometry in patients in euthymic, depression or hypomanic phase of the disease and compared with healthy controls. All BD patients were characterized by lower percentage of CD3+CD4+ and CD3+CD8+ cells compared with healthy people. But only patients in depression and remission had higher percentage of B cells (CD19+ cells) compared with healthy people. The percentage of CD4+CD25+ and CD8+CD25+ cells was decreased in patients in hypomanic phase compared with healthy control. Patients in remission were characterized by increased concentrations of IL-6 and IL-10 and decreased level of TNF in blood serum. Significant correlations between immunologic parameters and the results of Hamilton or Young scale have also been found. Our results demonstrate that there are significant differences in lymphocyte subpopulations which depend on the phase of the disease the patient is currently in.

Association of serum retinol and circulating inflammatory cells with disease activity in ulcerative colitis patients

Purpose The purpose of this study is to explore the association of serum retinol and number of circulating inflammatory cells and disease activity in patients with ulcerative colitis. Design/methodology/approach A total of 60 patients with ulcerative colitis were enrolled in a cross-sectional pilot study. Patients were recruited from specialized clinic of Tabriz University of Medical Sciences, Iran between April and August 2015. Mayo clinic index was used to assess clinical disease activity score. Blood samples were collected. Serum retinol was assessed using HPLC to determine vitamin A status. Complete blood count and lymphocyte phenotyping were performed by automated hematology analyzer and flow-cytometric analysis, respectively. Findings According to Mayo scoring, 68.33 per cent of patients had mild and 31.66 per cent had moderate or severe disease activity. About 43.33 per cent of patients were vitamin A deficient, with 23.33 per cent having moderate to severe deficiency (serum retinol &lt; 20 µg/dl). Lower levels of serum retinol and higher count and percentages of CD3+, CD8+ T cells and neutrophil to lymphocyte ratio were statistically associated with disease activity according to univariate analysis (p = 0.002, 0.037, &lt;0.001, 0.031, 0.002 and 0.039); however, in binary logistic regression, only lower levels of serum retinol were independently associated with disease activity with a OR of 0.564 (p = 0.021; 95 per cent CI 0.35-0.92). Originality/value Vitamin A deficiency was detected in this study population. Patients with moderate to severe disease activity demonstrated lower serum retinol, higher CD8+ T cells and neutrophil to lymphocyte ratio compared to patients with mild disease activity.