NK cells expansion <i>in vitro</i> is followed by loss of inhibitory KIR expression

NK cells are innate lymphocytes that are able to eliminate altered cells, which makes them promising for the immunotherapy of viral diseases and tumors. The NK cell population is characterized by high phenotypic and functional diversity. In particular, in the pool of highly differentiated NK cells in the presence of cytomegalovirus (HCMV), a population of adaptive cells can be formed, characterized by a high lifespan and high cytotoxicity. However, in order to carry out a cytotoxic reaction, a NK cell must undergo a licensing process, during which it acquires the expression of NKG2A and KIRs. Currently, there are many effective methods of NK cell accumulation for subsequent use in therapy, one of them is the stimulation with IL-2 and K562-mbIL21 feeder cells. Highly differentiated adaptive-like NK cells are able to expand in respond to such stimulation. However, the phenotype of actively expanding NK cells dynamically changes. Loss of inhibitory KIR expression during intense proliferation of NK cells may adversely affect their cytotoxic potential. This work shows that highly differentiated CD56dimNKG2C+ NK cells from HCMV-seropositive individuals have a high proportion of KIR2DL2/3+ cells. This may indicate a high stability of KIR receptor expression in this population. We have shown that CD56dimNKG2C+ clonal cultures obtained by stimulation with IL-2 and K562- mbIL21 are characterized by high stability of KIR2DL2/3 expression compared to NKG2C-negative and less differentiated CD56brightNKG2C+. Also, in heterogeneous cultures of adaptive NK cells precursors CD57- CD56dimNKG2C+, a higher expression level of KIR2DL2/3 was observed in comparison with NKG2C-negative cultures of CD57-CD56dimNKG2C-. Thus, the accumulation of NK cells upon stimulation with IL-2 and K562- mbIL2 feeder cells can lead to loss of expression of KIR receptors and a decrease in their functional activity. However, cultures of highly differentiated NK cells of HCMV-seropositive individuals CD56dimNKG2C+, as well as cultures of precursors of adaptive NK cells CD57-CD56dimNKG2C+, are characterized by a greater stability of KIR2DL2/3 expression. As a result, stimulation with IL-2 and K562-mbIL21 feeder cells can be used to accumulate adaptive-like cells and their progenitors with stable inhibitory KIR expression and high cytotoxic potential

Self-similarity and singularity formation in a coupled system of Yang-Mills-dilaton evolution equations

We study both analytically and numerically a coupled system of spherically symmetric SU(2) Yang-Mills-dilaton equation in 3+1 Minkowski space-time. It has been found that the system admits a hidden scale invariance which becomes transparent if a special ansatz for the dilaton field is used. This choice corresponds to transition to a frame rotated in the $\ln r-t$ plane at a definite angle. We find an infinite countable family of self-similar solutions which can be parametrized by the $N$ - the number of zeros of the relevant Yang-Mills function. According to the performed linear perturbation analysis, the lowest solution with N=0 only occurred to be stable. The Cauchy problem has been solved numerically for a wide range of smooth finite energy initial data. It has been found that if the initial data exceed some threshold, the resulting solutions in a compact region shrinking to the origin, attain the lowest N=0 stable self-similar profile, which can pretend to be a global stable attractor in the Cauchy problem. The solutions live a finite time in a self-similar regime and then the unbounded growth of the second derivative of the YM function at the origin indicates a singularity formation, which is in agreement with the general expectations for the supercritical systems.Comment: 10 pages, 5 figure

Umbilical cord blood as a promising source of NK cells for immunotherapy

Currently, a large number of studies on genetic modification of cord blood NK cells (UCB-NK) are carried out at both clinical and preclinical levels. Immunotherapy based on UCB-NK cells has great potential for antitumor therapy. However, despite having known several advantages over peripheral blood NK cells (PB- NK), including a high concentration in cord blood and low virulence rate, UCB-NK cells are predominantly characterized in the scientific literature as immature and low-functioning NK cells. In this work, we studied the phenotypic characteristics of UCB-NK cells and the possibility of stimulatory compensation of the decreased functional activity of UCB-NK cells. Our studies revealed UCB-NK cells can be characterized as poorly differentiated and weakly activated cells with high level of inhibitory receptor NKG2A and low level of activating receptor NKG2C and HLA-DR, accordingly with the literature data. Two types of stimuli were chosen to stimulate freshly isolated UCB-NK cells: 1) 100 units of IL-2; 2) combinations of 100 units IL-2 and K-562 feeder cells expressing membrane-bound IL-21 (K562-mbIL21). It was shown the degranulation (LAMP-1) and proliferative activity was higher than for parallel cultured ex vivo PB-NK cells under the same conditions for UCB-NK cells stimulated for 7 days with IL-2 + K562-mbIL21. Moreover, stimulation in the way of IL-2 + K562-mbIL21 seemed to be a more perspective way to obtain a large number of proliferatively active UCB-NK cells compared to stimulation with IL-2 only. Since genetic modification of NK cells is a promising way to improve the antitumor properties of NK cells, retroviral transduction procedure was performed to study of the stimulated UCB-NK cells. UCB-NK cells stimulated with IL-2 + K562-mbIL21 were transduced on day 8 of cultivation. In this study, we used targeted overexpression of the adaptor molecule DAP12, which is involved in the signaling of activating NK cell receptors. PB-NK cells and UCB-NK cells were transduced under the equal experimental conditions in same volume of viral particles. As a result, the transduction efficiency was found to be more than 4-fold higher for UCB-NK cells compared to PB-NK cells. Thus, UCB-NK cells appear to be a promising tool for further research in cancer immunotherapy

Интегральный способ прогнозирования риска панкреатита после эндоскопической ретроградной холангиопанкреатографии

The objective: development of a method for determining the individual risk of post-ERCP pancreatitis (PEP) before endoscopic intervention.Subjects and Methods. A prospective observational study of the results of therapeutic endoscopic retrograde cholangiopancreatography (ERCP) was performed in 1,210 patients. The primary end point was the development of PEP. Multiple regression logistic analysis was used to determine the risk of developing PEP.Results. A model of independent variables was determined, which significantly (p = 0.0001) affected the risk of developing PEP. Based on the results of the logistic analysis, regression coefficients were calculated for all significant patient-dependent factors (gender, age, preoperative level of amylasemia and bilirubinemia, and nature of the underlying disease). The risk of PEP decreased in men (by 47.5%), with an increase in the age of patients (for 1 year by 2.3%) and bilirubin (with an increase of 1 μmol/l by 0.3%) but increased with an increase in amylase activity blood serum (by 0.7% for each 1 U/l). A formula has been developed to assess the individual risk of PEP at the stage of preparing a patient for therapeutic ERCP.Conclusion. Analysis of patient-dependent factors makes it possible to reliably determine the risk of developing PEP. It was determined that a female gender, young age (under 40 years old), sphincter of Oddi dysfunction increased the risk of PEP, while a male sex, stenting of the main pancreatic duct, and jaundice reduced this risk. The use of the proposed formula will help predict a significant (p = 0.0001) degree of risk of PEP before performing primary therapeutic ERCP in a particular patient.Цель: разработка способа определения индивидуального риска развития пост-ЭРХПГ-панкреатита (ПЭП) до выполнения эндоскопического вмешательства.Материал и методы. Проведено проспективное наблюдательное исследование результатов лечебной эндоскопической ретроградной холан- гиопанкреатографии (ЭРХПГ) у 1 210 пациентов. Первичной конечной точкой считали развитие ПЭП. Для определения риска развития ПЭП применяли множественный логистический регрессионный анализ.Результаты. Определена модель независимых переменных, которая достоверно (p = 0,0001) влияет на риск развития ПЭП. По результатам логистического анализа вычислены регрессионные коэффициенты для всех значимых пациент-зависимых факторов (пол, возраст, предоперационный уровень амилаземии и билирубинемии, характер основного заболевания). Риск ПЭП снижался у мужчин (на 47,5%), при увеличении возраста пациентов (за 1 год на 2,3%) и уровня билирубина (при увеличении на 1 мкмоль/л на 0,3%), но повышался при увеличении активности амилазы сыворотки крови (на 0,7% на каждую 1 Ед/л). Разработана формула, позволяющая оценить индивидуальный риск ПЭП на этапе подготовки пациента к выполнению лечебной ЭРХПГ.Вывод. Анализ пациент-зависимых факторов позволяет достоверно определить риск развития ПЭП. Определено, что женский пол, молодой возраст (до 40 лет), дисфункция сфинктера Одди увеличивали, а мужской пол, стентирование главного панкреатического протока, наличие желтухи снижали риск развития ПЭП. Использование предложенной формулы поможет прогнозировать достоверную (p = 0,0001) степень риска ПЭП до выполнения первичной лечебной ЭРХПГ у конкретного пациента

Production of {\pi}+ and K+ mesons in argon-nucleus interactions at 3.2 AGeV

First physics results of the BM@N experiment at the Nuclotron/NICA complex are presented on {\pi}+ and K+ meson production in interactions of an argon beam with fixed targets of C, Al, Cu, Sn and Pb at 3.2 AGeV. Transverse momentum distributions, rapidity spectra and multiplicities of {\pi}+ and K+ mesons are measured. The results are compared with predictions of theoretical models and with other measurements at lower energies.Comment: 29 pages, 20 figure

ASSOCIATION OF LETHAL OUTCOMES IN SURGICAL SEPSIS WITH ALLELIC VARIANTS OF TNF Α AND HEAT SHOCK PROTEIN (HSP70-2) GENES

Abstract. Taking into account close localization of genes encoding HSP70-2 and TNFα and mutual influence of their products upon development of septic process, we investigated allelic polymorphism in the coding region of HSP70-2 gene at position 1267, and a promoter polymorphism of TNFα gene (–308 position) in sixty-one patients with surgical sepsis, as compared with 100 healthy donors, assuming that certain genomic variants can be associated with susceptibility for sepsis and/or fatal outcomes in severe sepsis. Significant differences in frequencies of HSP70-2 gene alleles have been found in the group with sepsis against healthy controls. Prevalence of HSP70-2*B/B allele is increased in the group of patients with sepsis, and a predisposal for sepsis development is shown for carriers of the given genotype. When analyzing carriers of combined HSP70-2/TNFα allelic variants among dead patients vs survivors, we have revealed specific combinations that are detected solely among non-survived patients, i.e., HSP70-2 A/TNFα AA, or only among survivors (HSP70-2BB /TNFα AA и HSP70-2 BB/TNFα AG). The results obtained allow us to suggest about certain genetic factors that may sufficiently influence intensity of inflammation in sepsis and, hence, upon clinical manifestations and outcomes of the disease

Endoscopic optic coherent tomography in diagnosis of early bladder cancer

Our goal was statistical assessment of the in vivo cystoscopic optical coherence tomography (OCT) ability to detect early neoplasia in human urinary bladder. We analyzed major reasons of false positive and false negative image recognition results. Optical coherence tomography was performed to image the bladder during cystoscopy. The study enrolled 114 zones with suspicion for bladder cancer. The diagnosis was established by histopathology examination of a biopsied tissue. Each biopsy site was examined by OCT. Two physicians blinded to all clinical data participated in the recognition (malignant, suspicion or benign) of the OCT images. 82% sensitivity and 85% specificity for the OCT recognition of dysplastic/malignant versus benign/reactive conditions of the bladder were demonstrated. The interobserver agreement multi-rater kappa coefficient is 0.56. Good sensitivity and specificity of the OCT method in the diagnostics of bladder neoplasia makes OCT a promising complementary cystoscopic technique for non-invasive evaluation of zones suspicious for early cancer