Peripheral Blood Lymphocyte Subpopulations Research Articles

SUBPOPULATIONS OF LYMPHOCYTES AND MONOCYTES IN PATIENTS` VENOUS BLOOD WITH ATRIAL FIBRILLATION OR ATRIAL FLUTTER ASSOCIATED WITH HYPERTENSION

Objective: The most common reason of arrhythmia onset nowadays is hypertensive desease. The aim of the study is to compare the subpopulations of lymphocytes (LCTs) and monocytes (MCTs) in blood of patients (pts) with paroxysmal and persistent forms of AF or atrial flutter (AFL) associated with arterial hypertension. Design and method: The study involved 103 patients with atrial fibrillation and flutter that occurred on the backdround of hypertension. Depending on the form of arrhythmia, patients were divided into three main groups: group I – with paroxysmal form of atrial fibrillation, group II – with a persistent form of atrial fibrillation, group III – with a persistent form of atrial flutter. The control groups included patients with hypertension, but without these arrhythmias and healthy individuals who entered groups IV and V, respectively. The lymphocytes and monocytes subpopulation in peripheral blood was assessed by flow cytometry. Results: Analyzing the lymphocyte subpopulations in peripheral blood of patients with atrial fibrillation and flutter (groups I, II and III), it was found that the number of cells with cytotoxic activity (NK and NKT) in both absolute count and percentage values was significantly higher than in healthy individuals. A statistically significant decrease of T-regulatory cells number was found in patients with arrhythmias compared to control groups (p < 0.05). In patients with AF and AFL associated with hypertension, compared to patients with hypertension without these rhythm disturbances or healthy individuals, there is an increased number of classical and intermediate monocytes subpopulations. Conclusions: 1. In patients with atrial fibrillation or atrial flutter associated with hypertension, compared to patients with hypertension without these rhythm disturbances or healthy people, there is an increased number of classical and intermediate monocytes fraction and a decreased content of T-regulatory cells in peripheral blood, an increased number of cells with cytotoxic activity (NK and NKT) compared to healthy individuals. These changes can be one of the factors for the activation of systemic and local inflammatory processes that can lead to fibrosis in the myocardium and the associated electrical and structural remodeling typical for AF and AFL.

Peripheral Th and NK cells are associated with response to lenvatinib plus PD-1 inhibitor in unresectable hepatocellular carcinoma.

The value of peripheral blood lymphocyte subpopulations in predicting responses to lenvatinib combination with programmed death-1 (PD-1) inhibitors in unresectable hepatocellular carcinoma (HCC) was investigated. Fifteen patients received objective responses (OR) and sixteen patients had non-objective responses (NOR) were analyzed. The counts of peripheral blood lymphocyte subpopulations from patients were measured before treatment, second (at week 3), and third doses (at week 6) of the PD-1 inhibitor administration, and correlated with responses. Helper T (Th) cells and natural killers (NK) cells were more abundant in the OR group and found to be important predictors of OR in a stepwise multivariate logistic regression analysis. These cutoff values of Th and NK cells could help to distinguish OR from NOR cases accurately and provide clinical benefits.

Morphofunctional changes in brain and peripheral blood in aged Wistar rats due to AlCl3 exposure

The purpose of the study: To examine morphofunctional changes of brain and peripheral blood in aged Wistar rats to observe adaptive reactions as a response on AlCl3 exposure. Methods: The work was performed on male Wistar rats, 24 months of age. Animals consumed a solution of AlCl3 100 mg/kg per day for 60 days. Morphological changes of neurons and microglia, mRNA expression levels of pro- and anti-inflammatory cytokines, microglia activation markers, amyloid-related and hypoxia-related proteins, as well as monocyte and lymphocyte subpopulations in peripheral blood, were examined. Results: AlCl3-treated old rats showed the increasing of hyperchromic neurons in 2 out of 3 examined regions of the hippocampus; morphological features of microglia cells’ dystrophy; the upregulation of pro-inflammatory cytokine Il-18 and the downregulation of anti-inflammatory cytokine Il-10, as well as App, Bace1, and Hif-1a; the decreasing of percentage of B-cells, general CD3+ lymphocyte population, including CD4+ T-helpers and CD8+ cytotoxic cells, but the increasing of CD4+/CD8+ ratio in peripheral blood. Conclusion: AlCl3-treated aged rats demonstrated systemic maladaptation to AlCl3 impact. Unlike adult rodents, aged ones have the background of inflammaging, as well as elderly people. The exposure of AlCl3 could potentially be a replacement of integral cellular and molecular processes accompanying age-related diseases, presenting in most elderly people, as an enhancer of inflammaging and hypoxia. These conditions make this model of neurodegeneration a reliable one to explore the initial mechanisms of such detrimental process, as well as prove aged rats more suitable subjects to perform future researches in this field.

Peripheral blood lymphocyte changes after stereotactic ablative body radiotherapy to lung or liver metastases in patients with oligometastatic cancers.

To perform the analysis of the peripheral blood lymphocyte changes after stereotactic ablative radiotherapy (SABR) in patients with oligometastatic cancers. The dynamics of the immune status in peripheral blood was prospectively evaluated in 46 patients with lung (17 cases) or liver (29 cases) metastases treated by SABR. Flow cytometry of peripheral blood lymphocyte subpopulations was performed before SABR, 3-4 weeks and 6-8 weeks after the end of SABR: 3 fractions of 15-20 Gy or 4 fractions of 13.5 Gy. The number of treated lesions varied from 1 (32 patients) to 2-3 (14 patients). SABR induced a significant increase of T-lymphocytes (CD3+CD19-) (p = 0.001), T-helper (CD3+CD4+) (p = 0.004), activated cytotoxic T-lymphocytes (CD3+CD8+HLA-DR+) (p = 0.001), activated T-helpers (CD3+CD4+HLA-DR+) (p < 0.001). A significant decrease of T-regulated immune suppressive lymphocytes (CD4+CD25brightCD127low) (p = 0.002) and NKT-cells (CD3+CD16+CD56+) (p = 0.007) was recorded after the SABR. The comparative analysis demonstrated that lower doses of SABR (EQD2Gy(α/β=10) = 93.7-105.7 Gy) induced significant increase of T-lymphocytes, activated cytotoxic T-lymphocytes, and activated CD4+CD25+ T-helpers, while SABR with higher doses (EQD2Gy(α/β=10) = 150 Gy) was not associated with these effects. A more efficient activations of T-lymphocytes (p = 0.010), activated T-helpers (p < 0.001), and cytotoxic T-lymphocytes (p = 0.003) were associated with SABR to a single lesion. A significant increase of T-lymphocytes (p = 0.002), T-helpers (p = 0.003), and activated cytotoxic T-lymphocytes (p = 0.001) was observed after SABR for hepatic metastases in contrast to SABR for lung lesions. Changes in peripheral blood lymphocytes after SABR could be influenced by the location or the number of irradiated metastasis, and the dose of SABR.

Половые различия в субпопуляционном составе лимфоцитов периферической крови при язвенном колите отражают специфические особенности иммунопатогенеза воспалительных заболеваний кишечника у мужчин и женщин

Objective. To identify the features of the composition of peripheral blood lymphocyte subsets in women with ulcerative colitis (UC). Patients and methods. The study included 32 women: 21 patients with UC and 11 healthy volunteers (control group) without inflammatory bowel disease (IBD). The major subsets of T and B lymphocytes were detected by multicolor flow cytometry. Results. The analysis of peripheral blood CD3+ T cell differentiation in women with UC in comparison with healthy controls showed that the Tcyt group had a significant decrease in naive Tcyt cells with an increase in the level of Tcyt cell subsets with pronounced effector properties (EM Tcyt and TEMRA Tcyt). In the subsets of central and effector memory Th cells in women with UC compared to healthy controls, Th17.1-CM cells were significantly different among central memory (CM) cells, and Th17-EM and Th17.1-EM cells were significantly different among effector memory (EM) cells. No significant differences were found in any of the Th17 cell subsets, but there was a tendency towards decreased levels of Th17.1 cells. The analysis of the subsets of T follicular helper (Tfh) cells revealed significant changes only in the level of Tfh17/Tfh22-EM cells (Tfh17-EM cell subset). No significant differences in the B cells were revealed, except for plasmocytes with IgD–CD27++ phenotype, which were increased in UC patients. Conclusion. Changes in peripheral blood T- and B-lymphocytes in women with UC can reflect not only activation of the peripheral immune system, but also induction of immunoglobulin production by B cells activated by T follicular helper cells. For the first time, specific differences between women and men in the total pool of Th17/Th22 cells, “double-positive” Th17, Tfh2 and Tfh17/Tfh22 cells were identified. In contrast to men, no manifestations of a Th2-specific response at the cellular level were observed in women with UC. The revealed sex-related differences in the composition of lymphocyte subpopulations in peripheral blood in UC reflect specific features of the IBD immunopathogenesis in men and women and can be considered in the management of patients with IBD. Key words: inflammatory bowel disease, ulcerative colitis, T helper cell subsets, B lymphocytes, sex-related differences

Subpopulations of lymphocytes and monocytes in blood of patients with atrial fibrillation or atrial flutter associated with hypertension

Atrial fibrillation (AF) remains one of the most common arrhythmias, second only to supraventricular extrasystoles, but the universal cause of its occurrence is still unknown. The inflammatory theory of arrhythmogenesis attracts the attention of researchers around the world. The purpose of the study is to compare the subpopulations of lymphocytes and monocytes in blood of patients with paroxysmal and persistent forms of atrial fibrillation or atrial flutter (AFL) that associated with arterial hypertension. Materials and methods. The study involved 103 patients with atrial fibrillation and flutter that occurred secondary to hypertension. Depending on the form of arrhythmia, they were divided into three main groups: group I – with paroxysmal form of atrial fibrillation, group II – with a persistent form of atrial fibrillation, group III – with a persistent form of atrial flutter. The control groups included patients with hypertension, but without these arrhythmias and healthy individuals who entered groups IV and V, respectively. The lymphocytes and monocytes subpopulation was assessed by flow cytometry in peripheral blood. Results and discussion. Analyzing the lymphocyte subpopulations in peripheral blood of patients with atrial fibrillation and flutter (groups I, II and III), it was found that the number of cells with cytotoxic activity (NK and NKT) in both absolute count and percentage values was significantly higher than in healthy individuals. A statistically significant decrease of T-regulatory cells number was found in patients with arrhythmias compared to control groups (p ≤ 0.05). In patients with AF and AFL associated with hypertension, compared to patients with hypertension without these rhythm disturbances or healthy individuals, there is an increased number of classical and intermediate monocytes subpopulations. Conclusions. In patients with atrial fibrillation and atrial flutter that occurred as a result of hypertension, compared to patients without arrhythmias or healthy people, there is an increased content of pro-inflammatory subpopulations of blood monocytes, T-cytotoxic cells and a decrease in the content of T-regulatory cells.

Correlation between peripheral blood lymphocyte subpopulations and primary systemic lupus erythematosus.

This study explored the correlation between peripheral blood CD3＋, CD3＋/CD4＋, CD3+/CD8+, CD4＋/CD8＋, CD3−/CD16+ CD56+, and CD3−CD19+ and disease activity of different subtypes of systemic lupus erythematosus (SLE). The percentages of CD3+, CD3+/CD4+, CD3+/CD8+, CD4+/CD8+, CD3−/CD16+ CD56+, and CD3−CD19+ in the peripheral blood of patients (n = 80) classified into lupus nephritis, blood involvement, and joint involvement and SLE in different active stages were detected by flow cytometry. Their correlations with baseline clinical experimental indicators of SLE patients’ SLE disease activity index score (SLEDAI) and complement C3 were analyzed. The results showed that CD3+, CD3+/CD4+, and CD3+/CD8+ at baseline level were negatively correlated with SLEDAI scores. These were positively correlated with C3. In conclusion, T-lymphocyte subpopulations are closely related to SLE activity and can be used as reference indicators to evaluate the SLE activity.

Application of the diagnostic criteria for Common Variable Immunodeficiency in resource-limited settings.

Common variable immunodeficiency (CVID) is the most prevalent symptomatic humoral deficiency; however, its heterogeneous presentation makes the diagnosis difficult. The present study is aimed to verify the CVID diagnostic criteria as established by the European Society for Immunodeficiencies in 42 CVID patients from our outpatient clinic. Information was collected from their medical records and when needed, lymphocyte subpopulations in peripheral blood (PB) were performed by flow cytometry. All the patients fulfilled the clinical working definition for CVID and showed decreased serum IgG and IgA at diagnosis. Over two-thirds of the patients had decreased memory B cell percentages. However, the remaining patients exhibited other quantitative B cell defects in PB. Evaluation of vaccination responses was only found in 13 records and 69% were not responsive. None of the patients were subjected to vaccination studies to both, T-cell dependent and independent antigens. The two required tests to evaluate T cell responses were performed in 84.2% of the patients and reported normal. Without the support of third-party payers, only 34.2% of our patients would have completed the required evaluations. Further efforts are needed to speed up CVID diagnosis in low-resourced settings, increasing the availability of the required resources and optimizing the healthcare supply chain.

PB2276: EARLY ADMINISTRATION OF REMDESIVIR PLUS CONVALESCENT PLASMA THERAPY IS EFFECTIVE TO TREAT COVID-19 PNEUMONIA IN B-CELL DEPLETED PATIENTS WITH HEMATOLOGICAL MALIGNANCIES

Background: Patients with hematological malignancies are at a higher risk of developing severe form and protracted course of COVID-19. Remdesivir, an inhibitor of RNA polymerase was identified early as a promising therapeutic regimen for COVID-19, while convalescent plasma (CP) therapy with anti-SARS-CoV-2 antibodies can aid the treatment of COVID-19 in B-cell depleted patients. Aims: Here we investigated whether the combination of externally administered immunoglobulins (Ig) via CP therapy and specific inhibition of viral replication might be sufficient to effectively treat B-cell depleted patients with COVID-19. Methods: We enrolled a series of 20 consecutive patients who suffered from various hematological malignancies with profound B-cell lymphopenia and COVID-19 pneumonia between December 2020 and May 2021. These patients demonstrated a profound B-cell lymphopenia based on flow cytometry analysis of peripheral blood lymphocyte subpopulations. Consequently, they showed undetectable baseline anti-SARS-CoV-2 Ig levels before CP therapy according to anti-SARS-CoV-2 nucleocapsid- and S1-RBD-specific total Ig tests. Each patient received a complete course of remdesivir and at least one unit (200 ml) of AB0 compatible convalescent plasma during their treatment for COVID-19. Results: Previous anti-CD20 therapy resulted in a more prolonged SARS-CoV-2 PCR positivity compared to other causes of B-cell lymphopenia (p=0.004). Timing of CP therapy showed a significant impact on the clinical outcome. Simultaneous use of remdesivir and CP reduced time period for oxygen weaning after diagnosis (p=0.017), length of hospital stay (p=0.007), and PCR positivity (p=0.012) compared to patients who received remdesivir and CP consecutively. In addition, time from the diagnosis to CP therapy affected the length of oxygen dependency (p<0.001) and hospital stay (p<0.0001). In those cases where there were at least ten days from the diagnosis to plasma administration, oxygen dependency was prolonged vs. patients with shorter interval (p<0.001). Summary/Conclusion: Via combined administration of remdesivir and CP, we experienced an overwhelming survival rate in COVID-19 patients with hematological malignancies. Inhibition of viral replication with passive immunization proved to be efficient and safe. Our results suggest the clear benefit of early administration of CP to avoid protracted COVID-19 disease in patients with B-cell lymphopenia.

Innate and Acquired Cellular Immunity in Children with Familial Hypercholesterolemia Treated with Simvastatin.

The aim of this cross-sectional study was to assess the influence of simvastatin treatment in children with familial hypercholesterolemia (FH) on parameters of cellular immunity. Twenty-six children with FH were included, of which thirteen were treated with 10 mg simvastatin for at least 26 weeks, and thirteen were age- and sex-matched with a low-cholesterol diet only. Total WBC count and lipid profile were measured. Flow cytometry was used to identify lymphocyte subsets and determine the expression of adhesion molecules (AM) and toll-like receptors (TLRs) on leukocytes. No differences were found in the basic values of peripheral blood count and subpopulations of lymphocytes between groups. The percentage of granulocytes with the expression of AM was higher in those treated with statins. The TLR-2 expression on granulocytes and monocytes showed higher values, whereas the TLR-4 expression was lower on lymphocytes and granulocytes in simvastatin-treated children. Treatment with simvastatin in children with FH is not associated with alterations in the amounts of granulocytes and monocytes. There is no association between statin treatment and the pattern of peripheral blood lymphocyte subpopulations. The role of AM and TLRs needs further investigation, given the effect of statins on the innate immunity may be important for their efficacy and safety during growth.

Lymphocyte Subsets in Defense Against New Pathogens in Patients with Cancer.

Immunity in cancer patients is modified both by the cancer itself and by oncospecific treatments. Whether a patient's adaptive immunity is impaired depends on their levels of naive lymphocytes and other cell populations. During the COVID-19 pandemic, cancer patients are at greater risk of progressing to severe forms of the disease and have higher mortality rates than individuals without cancer, particularly while they are receiving cancer-specific therapies. An individual's protection against infection, their response to vaccines, and even the tests that determine the humoral immune response to SARS-CoV-2, depend on lymphocyte populations, meriting their study. Estimate blood concentrations of lymphocytes involved in the immune response to new pathogens in cancer patients. We carried out an analytical study of 218 cancer patients; 124 women and 94 men, 26-93 years of age, who were treated at the National Oncology and Radiobiology Institute in Havana, Cuba, March-June, 2020. Patients were divided into five groups: (1) those with controlled disease who were not undergoing cancer-specific treatment; (2) those undergoing debulking surgery; (3) patients undergoing chemotherapy; (4) patients undergoing radiation therapy and (5) patients currently battling infection. We evaluated the following peripheral blood lymphocyte subpopulations via flow cytometry: B lymphocytes (total, naive, transitional, memory, plasmablasts and plasma cells); T lymphocytes (total, helper, cytotoxic and their respective naive, activated, central memory and effector memory subsets); and total, secretory and cytotoxic natural killer cells and T natural killer cells. We also estimated neutrophil/lymphocyte ratios. Lymphocyte concentrations were associated with controlled disease and standard cancer therapy. For variables that did not fall within a normal distribution, ranges were set by medians and 2.5-97.5 percentiles. The two-tailed Mann-Whitney U test was used to measure the effect of sex and to compare lymphocyte populations. We calculated odds ratios to estimate lymphopenia risk. All cancer patients had lower values of naive helper and cytotoxic T lymphocyte populations, naive B lymphocytes, and natural killer cells than normal reference medians. Naive helper T cells were the most affected subpopulation. Memory B cells, plasmablasts, plasma cells, activated T helper cells, and cytotoxic central memory T cells were increased. Patients undergoing treatment had lower levels of naive lymphocytes than untreated patients, particularly during radiation therapy. The risk of B lymphopenia was higher in patients in treatment. The odds ratio for B lymphopenia was 8.0 in patients who underwent surgery, 12.9 in those undergoing chemotherapy, and 13.9 in patients in radiotherapy. Cancer and conventional cancer therapies significantly affect peripheral blood B lymphocyte levels, particularly transitional T helper lymphocytes, reducing the immune system's ability to trigger primary immune responses against new antigens.

Secondary Immunosuppression in Pediatric Kidney Transplant Recipients.

Currentimmunosuppressive treatments for kidney transplant recipients have improved graft viability at the expense of impaired immune surveillance. The tools for monitoring immune status in pediatric kidney transplant recipients have not been widely investigated. Better knowledge could help recognize over immunosuppression and allow implementation of individualized preventive strategies. This retrospective and observational study included 28 pediatric kidney transplant recipients treated at a tertiary hospital. We measured peripheral blood lymphocyte subpopulations, immunoglobulins, immunosuppressivedrug levels, and viral loads. Reference analytical values for different age ranges were used to determine immune status. We recorded overall hospitalizations due to opportunistic infections and positive viral loads posttransplant. We found hypogammaglobulinemia and lymphopenia in 19% and 41% of the patients, respectively. Peripheral blood lymphocyte subpopulations were below normal limits in one-third of the sample. These parameters were not related to the current number or plasma levels of immunosuppressive drugs. During follow-up, cytomegalovirus, Epstein-Barr virus, and BK virus viremias were detected in 60.7% of the patients. Admissions due to opportunistic infections happened in 57.1%, mainly related to severe viral disease (30%) or gastrointestinal infections (26.7%). Most occurred in younger transplant recipients and during the first 2 years posttransplant (73.3%). We found no significant relation between peripheral blood lymphocyte subpopulations and hospital admissions for opportunistic infections or positive viral loads during follow-up. Recurrent hospitalizations for opportunistic infections and analytical disorders in the immune system suggested that secondary immunosuppression in pediatric kidney transplant recipients was frequent. Immunosuppression was not directly related to plasma drug levels or the number of immunosuppressive drugs. Thus, immune monitoring might be helpful in combination with immunosuppressant levels to assess immunosuppression status and to establish individualized preventive measures.

Main lymphocyte subpopulations in cerebrospinal fluid and peripheral blood in HIV-1 subtypes C and B.

HIV-1 subtype C (HIV-1C) shows reduced Tat protein chemoattractant activity compared with HIV-1B. The impact of HIV-1C Tat on the chemotaxis of the main lymphocyte subpopulations in the cerebrospinal fluid (CSF) and the peripheral blood (PB) is unclear. We hypothesized that there would be a lower frequency of specific lymphocyte subpopulations CD3+ or CD19+ in CSF in HIV-1C than in HIV-1B. The objectives were to detect the differences in the proportions of main lymphocyte subpopulations in CSF and PB, between people with HIV (PWH) and HIV-1-uninfected volunteers (PWoH) and in HIV-1B and HIV-1C. Lymphocyte immunophenotyping was studied in CSF and paired PB samples of PWH (n = 22) and PWoH (n = 14). Lymphocytes were analyzed within the CD45+ gated region. The proportions of CSF CD3+CD4+, CD3+CD8+, and CD3-CD19+ lymphocytes in CSF were comparable in HIV-1B and C. There was an increase in the proportion of CD3+CD8+ cells and a decrease in CD3+CD4+ T cells (ps = 0.016) in the CSF samples of the PWH compared with the PWoH group. In the PWH group, both CD3+CD4+ and CD3+CD8+ lymphocytes were significantly higher in the CSF than in the PB (p = 0.047 and 0.005). The proportion of CD3+CD4+ was lower and that of CD3+CD8+ was higher in the CSF samples of the aviremic group than that of HIV-negative control (p = 0.0008 and < 0.0001, respectively). HIV-1C Tat substitution (C30S) did not interfere with the CNS migration of the main lymphocyte subpopulations. This is the first study to evaluate these lymphocytes in CSF and PB of HIV-1C compared with HIV-1B.

Prospective study of the cytokine profile and antioxidant status of patients with pyoderma

Background. The importance of studying the pathogenetic features of pyoderma associated with Streptococcus pyogenes is associated with the severity of the course of the disease, frequent relapses with a short inter-relapse period, and long-term disability.The severity of the course, long-term disability, violations of the pro-oxidate mechanisms recorded in patients with poidermias associated with Streptococcus pyogenes are priority and timely.&#x0D; Aims. Study of the cytokine, prooxidant and antioxidant profiles of patients with pyoderma associated with Streptococcus pyogenes.&#x0D; Мethods. А prospective study of the cytokine profile, the level of antioxidant defense enzymes, and lipid peroxidation factors in patients with pyoderma, lasting more than 2 years, associated with Streptococcus pyogenes, was carried out. The study included 100 people with diffuse lesions of smooth skin, identified on the skin surface of Streptococcus pyogenes, at the age of 28.76 6.24 years; in blood serum, the content of cytokines IL-2, IL-8, IL-10, TNF-, IFN-, IL-1 was studied by enzyme immunoassay, in whole blood the content of primary, secondary and final products of lipid peroxidation, antioxidant enzymes protection superoxide dismutase and catalase.&#x0D; Results. Analysis of the content of cytokines showed a significant decrease in the concentration of IL-2, an increase in IL-10, TNF-, IL-8, IL-1 in the blood serum of patients with streptoderma compared with the reference values of healthy people (p 0.05). In whole blood plasma, an increase in the concentration of primary, secondary and final products of lipid peroxidation was registered with a significant decrease in the activity of antioxidant enzymes of superoxide dismutase and catalase relative to the indicators of healthy volunteers (p 0.05).&#x0D; Conclusions. In patients with streptoderma, an increase in the production of cytokines with pro-inflammatory and chemotactic properties in response to the invasion of the pathogen and its persistence was revealed. The intracellular nature of the parasitization of Streptococcus pyogenes contributes to the enhancement of the systemic and local inflammatory process, the formation of oxidative stress, the accumulation of primary, secondary and final products of lipid peroxidation, and a decrease in the activity of antioxidant enzymes. A decrease in the content of interleukin 2, which has immunoregulatory properties in patients with streptoderma, leads to impaired immune homeostasis, a decrease in the processes of differentiation in the direction of the Th1 immune response, proliferation of T-lymphocytes, a violation of the qualitative and quantitative composition of the main subpopulations of peripheral blood lymphocytes, which is clinically expressed in this occurrence of relapses in categories of patients.

Herpes Virus Reactivation in Lymphoma Patients During and After Autologous Hematopoietic Stem Cell Transplantation

Aim. To assess the detection rate of human herpes virus DNA (of cytomegalovirus, herpes simplex virus types 1 and 2 [HSV-1/2], human herpes virus type 6 [HHV-6], and Epstein-Barr virus) in different biological environments at different stages of autologous hematopoietic stem cell transplantation (auto-HSCT) as well as the effect of immune factors on reactivation of viruses under study. Materials &amp; Methods. From 2019 to 2021 the study enrolled 87 lymphoma patients during and after auto-HSCT. Virological monitoring was performed on biological fluids (blood, saliva, urine, etc.) prior to conditioning regimen on Day 0 as well as on Day +5 and Day +10 after auto-HSCT. On these days (Day 0, Day +5, and Day +10) the immune factors (IgM, IgG, and IgA levels and pattern of lymphocyte subpopulation in peripheral blood) in 15 % (14/87) of patients were assessed in terms of their effect on herpes virus reactivation. Results. The overall rate of viral DNA detection increased from 26 % (26/87) to 42 % (37/87) of cases in the period of granulocytopoietic recovery. The most frequent were HHV-6 and HSV-1/2 reactivations reported in 23 % (20/87) and 16 % (14/87) of cases, respectively. The median B-lymphocyte proportion in peripheral blood of patients with herpes virus reactivation was 0.26 %, whereas in patients without reactivation it was 6.7 % (p = 0.019). The median absolute B-lymphocyte count in the cohort of patients with detected viral DNAs was 0.001 &lt;sup&gt;x&lt;/sup&gt; 10&lt;sup&gt;9&lt;/sup&gt;/L, whereas in patients without them it was 0.098 &lt;sup&gt;x&lt;/sup&gt; 10&lt;sup&gt;9&lt;/sup&gt;/L (p = 0.026). Conclusion. A high rate of herpes virus DNA detection in lymphoma patients after auto-HSCT affected neither transplant engraftment nor transplantation mortality. Immune predictors of virus infection reactivation were the decreasing proportion of B-cells in the total lymphocyte count and the absolute B-lymphocyte count in the peripheral blood prior to auto-HSCT.

Features of the immune system cells in abdominal obesity and metabolic syndrome and their associations with factors of cardiometabolic risk, visceral adipose tissue dysfunction and adipokines

The aim of this work was to establish the features of the immune system cells in abdominal obesity (AO) and metabolic syndrome (MS) and to determine the associations between the studied parameters and MS components, insulin resistance, and the level of adipokines. 244 patients of both sexes aged 18–45 years were included. The patients were divided into 4 groups: group 1 (comparison group) — healthy individuals without AO, excess body weight and additional MS components (n=71); group 2 (AO) — patients with isolated AO (n=32); group 3 (AO+1) — patients with AO and 1 component of MS (n=63); group 4 (MS) — patients with MS (n=78). The laboratory investigation included determination of parameters of carbohydrate, lipid metabolism, adipokines. The calculation of insulin resistance indices and the index of dysfunction of visceral adipose tissue were performed. The parameters of cell immunity were studied (the number of leukocytes and leukocyte formula; the subpopulations of lymphocytes — CD3+45+, CD3+4+, CD3+8+, CD3+56+, CD3-56+, CD3-19+, CD3+25+ and CD3+HLADR+ ). The results of this work demonstrate inflammatory changes in the leukocyte formula, as well as the appearance of activated lymphocytes in peripheral blood in patients of groups 2–4. Correlation analysis results suggest that the mechanism of formation of changes in cell adaptive immunity in AO and MS is associated with cardiometabolic risk factors. Conclusions. 1. Changes in the leukocytes composition in peripheral blood in patients with AO and MS are an increase in the number of neutrophils, monocytes and a decrease in the number of lymphocytes. In the groups AO+1 and MS these changes are accompanied by an increase in the number of leukocytes. 2. The subpopulations of peripheral blood lymphocytes in the groups AO and AO+1 are characterized by an increase in the level of CD3+HLADR+. In patients with MS — in combination with an increase in the level of CD3+25+ and B-lymphocytes. 3. The appearance of activated lymphocytes in the peripheral blood in the groups AO and MS is associated with cardiometabolic risk factors (excess adipose tissue, dyslipidemia, hyperleptinemia, hyperinsulinemia, and insulin resistance).

Effect of vitiligo treatment by compound Glycyrrhizin combined with fractional laser and Triamcinolone Acetonide injection on T Lymphocyte subpopulation.

ABSTRACTObjectives:To discuss the effective mechanism of vitiligo treatment by compound glycyrrhizin combined with fractional laser and triamcinolone acetonide injection.Methods:Forty-two patients with vitiligo vulgaris in the stable phase were classified into combined group (19 cases) and medicine group (23) admitted in dermatology department, Baoding First Central Hospital from January 2017 to July 2018. Both groups took 50mg compound glycyrrhizin orally three times per day, and applied halometasone cream externally once per day. Based on this treatment method, after the combined group adopted fractional laser, triamcinolone acetonide injection encapsulation was used immediately. After the treatment for six months, the curative effect was judged for both groups. Flow cytometry was used to test the changes of T lymphocyte subpopulation in peripheral blood before and after treatment. Meanwhile, immunohistochemical method was adopted to determine CD4 + and CD8 + T lymphocyte expression level. Besides, the normal control group was set up.Results:The efficacy of combined group and medicine group were 73.68% and 56.52% respectively, P<0.05. The comparison of CD3+, CD4 +, CD8 + and CD4 +/CD8 + T lymphocyte level in serum and skin damage before and after treatment had no statistical significance (P>0.05). Serum CD4 + T cells of vitiligo patients reduced, compared with the normal control group (P<0.05), and CD4 +/CD8 + declined (P<0.05). CD4 + and CD8 + T lymphocytes at the skin damage of patients increased, compared with normal control group (P<0.05).Conclusions:Compound glycyrrhizin combined with fractional laser and triamcinolone acetonide injection has good clinical effect in the treatment of vitiligo vulgaris in the stable phase, and its effective mechanism may have nothing to do with T lymphocyte subpopulation.

Визначення рівня субпопуляцій лімфоцитів периферичної крові для діагностики хронічної реакції відторгнення ниркових алотрансплантатів

Актуальність. Однією з основних причин втрат ниркових алотрансплантатів (НАТ) у віддалені строки після трансплантації є прогресуюча хронічна дисфункція ниркового алотрансплантата (ХДНА), причиною розвитку якої в 20–60 % випадків є хронічне відторгнення (ХВ). Діагностика причин дисфункції трансплантата базується на морфологічному дослідженні, яке через інвазивність має ряд недоліків. Тому великий інтерес являє пошук неінвазивних методів діагностики стану НАТ, зокрема, дослідження клітинної ланки імунної відповіді. Мета. Дослідження відносної кількості субпопуляцій лімфоцитів периферичної крові та їх співвідношення в реципієнтів у віддаленому періоді після трансплантації нирки, оцінювання інформативності цих показників для діагностики ХВ НАТ. Матеріали та методи. Досліджено рівні субпопуляцій лімфоцитів (Т-клітини, Т-хелпери, Т-цитотоксичні, Т-активовані, T-NK-клітини, В-клітини, NK-клітини) у 43 реципієнтів НАТ залежно від перебігу віддаленого післяопераційного періоду та стану алотрансплантата через 1–9 років після алотрансплантації нирки, які були розподілені згідно з морфологічною верифікацією діагнозу на дві групи. Група 1 — 23 хворі із задовільною функцією НАТ без ознак відторгнення; група 2 — 20 хворих із ХДНА, обумовленою хронічною реакцією відторгнення (ХРВ) за даними пункційної біопсії. До контрольної групи увійшли 23 здорові донори, чиї показники становили референтний коридор. Результати. Дані аналізу показали підвищення середніх рівнів Т-активованих лімфоцитів у групах 1 та 2 щодо контрольної (8,9 ± 2,0; 8,4 ± 2,4; 6,8 ± 2,1 відповідно). Також виявлено зниження середніх показників рівнів T-NK-клітин (2,4 ± 2,0) та В-клітин (9,6 ± 2,0) у групі 2 щодо показників групи 1 (5,6 ± 1,8 та 12,4 ± 2,4 відповідно). Аналіз індексів співвідношення між окремими субпопуляційними одиницями лімфоцитів показав статистично вірогідну значущість різниці між індексом Т-клітини/T-NK-клітини (22,53 vs 46,89 в групах 1 та 2 відповідно; p &lt; 0,05) та індексом Т-активовані/В-клітини (1,14 vs 4,17 в групах 1 та 2 відповідно; p &lt; 0,05). При цьому чутливість індексу Т-клітини/T-NK-клітини становила 70 %, специфічність — 60 %. Чутливість індексу Т-активовані/В-клітини дорівнювала 55 %, специфічність — 50 %. Висновки. Зниження рівня T-NK-клітин ближче до нижнього порогового значення референтного коридора і, відповідно, співвідношення Т-клітини/T-NK-клітини в межах 44,49–49,29 (46,89 ± 2,40) у віддаленому післятрансплантаційному періоді може бути додатковою ознакою ХРВ як причини прогресуючої дисфункції НАТ.

Predictive biomarkers of inhibitors immune checkpoints therapy in malignant tumors

Immune checkpoint inhibitors (ICT) therapy is a successful immunotherapy (IT) strategy that is quite effective in a number of patients with non-small cell lung cancer, melanoma, bladder cancer, breast cancer and others. Nevertheless, there is a need in predictive markers for ICT therapy for personalized IT as far as there is a large group of patients, the proportion of which varies depending on the tumor, who do not have a clinical response to such therapy. The review summarizes the theoretical aspects and results of clinical trials dedicated to various clinical efficiency predictor using modern databases. As a result of the analysis it is established that the main candidates for the role of such markers are tumor infiltrating lymphocytes and their subpopulations, peripheral blood lymphocytes (PBL) and their subpopulations. PD1 (programmed death receptor 1) and PDL1 (programmed death receptor ligand 1) expression in tumor tissue can also be important for predicting IT efficiency. The most promising predictive biomarker meaning the most clinically relevant is a combination of the PBL subpopulations study and PD1 and PDL1 expression on the tumor cells.PubMed, Scopus, Web of Science, eLibrary, Russian Science Citation Index databases were searched for the available appropriate literature reports. The authors included 82 in the given review.

Low Circulating B Lymphocytes in Newly Diagnosed Follicular and Diffuse Large B Cell Lymphoma: Differences Compared to Normal Subjects and Possible Prognostic Role

Introduction: Reduction of the lymphocyte to monocyte ratio (LMR) has an unfavorable impact on overall survival in Diffuse Large B Cell Lymphoma (DLBCL), and on treatment response in Follicular Lymphoma (FL). Abnormal LMR seems primarily due to a reduction in absolute value of lymphocytes. Based on these observations, we have prospectively investigated peripheral blood (PB) and Bone Marrow (BM) lymphocyte subpopulations in a series of previously untreated DLBCL and FL. Aim of the study was: i. to assess whether LMR alteration is linked to a distinct lymphocyte subset; ii. to correlate PB with BM cell subpopulations; iii. to compared DLBCL and FL with regards to PB and BM lymphocyte subpopulations. Methods: Overall, PB cells from 58 DLBCL and 27 FL at diagnosis were investigated; BM cell populations were analyzed in 77 DLBCL and 54 FL. None of the studied patients had BM involvement nor circulating lymphoma cells detectable by cytometry; in addition, none of the patients had been previously treated with immunosuppressive drugs. PB from 40 healthy subjects and BM samples from 69 subjects without primary BM dysfunctions were used as control. Multicolor flow cytometry (MFC) was used to evaluate percentages and absolute values of the following cell subsets: CD3+ T cell, CD3-CD56+ NK cells, CD19+CD20+ B cells. Among B cells, t he analysis included transitional (CD38+high/CD24+high/IgD+/CD27-), naive (CD38+/CD24+/IgD+/CD27-), memory IgD+ (CD38-/+CD24+/IgD+/CD27+), and memory IgD- (CD38-/CD24+high/IgD-/CD27+) subsets; B cell precursors were investigated as CD19+CD20-CD10+ and mature B cells as CD19+CD20+CD10-. Results: A significant reduction in median absolute lymphocyte count was seen in PB from lymphoma patients (median: 1,526/µL) compared to controls (median: 2,195/ µL). Moreover, absolute lymphocytes were more markedly reduced in DLBCL (median: 1,386 /µL) compared to FL (median: 1883/µl) (p<0.001). This resulted in a significantly lower median PB-LMR in DLBCL (median LMR: 3) compared to FL patients (median LMR: 5) (p=0.001). Among PB cell subpopulations, a marked reduction in circulating B cells was observed in lymphoma patients (median 96/µL) compared to controls (median 224/µL). Again, loss of circulating B-cells was more markedly pronounced in DLBCL (median: 81.5 cell/µL) compared to FL (median: 150 cell/µL) (p=0.013). Differences in circulating B-cells among DLBCL, FL and controls are shown in Figure 1A. At BM analysis, a reduction of B cell precursor (CD19+/CD20-/CD10+) could be documented only in DLBCL. A reduction in circulating T-lymphocytes in lymphoma patients compared to controls was detected as well, although at a lower extent compared to B-cells. No significant abnormalities in the amount of circulating NK-cells was seen in both DLBCL and FL patients. When these data were matched with treatment outcome, the reduction of circulating B cells proved to be associated with an unfavorable outcome in younger patients with DLBCL. Indeed, among DLBCL patients aged < 60 yrs, a poorer progression free survival was seen in those having less than 96 B-cells/µL compared to those with PB B-cells equal or above 96 B-cells/µl (Figure 1B). Conclusions: The study indicates that circulating B lymphocytes are significantly reduced in both DLBCL and FL compared to age-matched healthy controls; however, their loss is much more pronounced in DLBCL compared to FL. Indeed, abnormalities in B-cell progenitors are present in BM from DLCBL but not from FL. In addition, B-cell reduction may have an unfavorable impact in the long-term outcome of DLBCL, at least in younger patients. Lastly, both B and T-cell subsets are decreased in DLBCL and FL whereas circulating NK subset does not seem to be affected. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.