Chronic EBV Infection Research Articles

АНАЛИЗ КЛИНИЧЕСКОГО СЛУЧАЯ ВУЛЬГАРНОЙ ПУЗЫРЧАТКИ, АССОЦИИРОВАННОЙ С ХРОНИЧЕСКОЙ ВЭБ-ИНФЕКЦИЕЙ (ГЛОССИТА, ОБУСЛОВЛЕННОГО ВЭБ)

Background. One of the autoimmune diseases is pemphigus, a chronic malignantly occurring autoimmune dermatosis accompanied by the development of blisters on the skin and mucous membranes. Isolated lesions of the SOPR occur in 10–20% of cases, complicating differential diagnosis. Numerous studies have proven the role of herpes virus infection as an aggravating factor in the course of true acantholytic pemphigus. The aim of the study was to determine the features of the clinical course of vulgar pemphigus against the background of reactivation of chronic EBV infection (glossitis caused by EBV). The clinical case of a patient with a diagnosis of “Vulgar pemphigus. Glossitis caused by the Epstein-Barr virus”. The anamnesis of the disease was carefully collected, a clinical and laboratory examination was performed with a cytological examination of a smear taken from a LITTER, a general blood test with an expanded leukocyte formula, C-reactive protein. In order to diagnose the viral load, PCR diagnostics was performed on the DNA of the herpes virus 1, 2, 4, 5 types in saliva, on the DNA of EBV in the blood, quantitatively. Specific antibodies of Ig classes M and G to EBV were determined by the ELISA method. Results. Due to the lifelong persistence of EBV in the structures of the oral cavity, its reactivation leads not only to chronic pastoral infection and the local development of the corresponding glossitis, but also, first of all, to a decrease in the level of local immunity. This fact aggravates the course of pemphigus, shortens the period of absence of relapse. The appointment of antiviral therapy at the first visit allowed to stabilize the symptoms caused by EBV, which led to positive dynamics in the course of pemphigus against the background of average dosages of prednisone in a shorter time. Conclusions. With clinical and laboratory confirmation of chronic EBV infection (glossitis caused by EBV), it is recommended to include antiviral and immunocorrective drugs, systemically and topically, in the complex treatment of pemphigus erythematosus.

Clinical experience in the use of local antiviral therapy for glossitis caused by the herpes virus type IV

Herpetic infections are one of the most common anthroponotic diseases. The prevalence of herpes virus types, their combined manifestation in the body of one person, as well as the increase in the number of people with secondary immunodeficiency make the problem of diagnosis and treatment of herpes-associated diseases the most urgent [1,2].As is known, herpes virus type IV (Epstein-Barr, EBV) is one of the frequently encountered members of the herpesvirus family [3]. According to the researches of many authors, by the age of 45 they have infected up to 100% of the world's population [4,5]. Persisting in the tissues of the mucous membrane of the oral cavity (lymphoepithelial pharyngeal ring of Pirogov-Valdeier, stroma of salivary glands, epithelial cells), reactivation of the virus occurs in the case of a decrease in immunity. [8,9]. At the same time, the number of diseases caused by EBV increases annually and depends on the phase of virus activity [10,11].One of the relatively "young" EBV-associated diseases is the corresponding glossitis, the clinical and laboratory characteristics of which are poorly presented in the literature. For this reason, the therapy of oral manifestations of chronic EBV infection is considered from the position of treating infectious mononucleosis [6,7], which is ineffective. Ignorance of dentists about the reliable clinical and laboratory signs of EBV caused by glossitis leads to incorrect diagnosis and inadequate treatment, generalization of infection, and the appearance of opportunistic diseases.

Minor population of NK lymphocytes with CD19 coexpression

Single reports were published concerning a minor subpopulation of NK cells with weak coexpression of the B cell antigen CD19 in the patients’ blood and bone marrow. The frequency and relative number of CD56+CD19+dim cells is virtually not assessed, and there is no data on their phenotypic characteristics, as well as the connection of this subpopulation with any disease state. The purpose of the present study was to assess the frequency, relative quantity and phenotypic characteristics of CD56+CD19+dim lymphocytes in blood of patients referred for assessment of the lymphocyte subpopulation profile. Peripheral blood of immunocompromised individuals was taken, and subpopulation composition of lymphocytes was determined using eight-color flow cytometry (markers: CD3, CD4, CD8, CD19, CD25, CD45, CD56, HLA-DR). To estimate incidence of the CD56+CD19+dim subpopulation, we have carried out a retrospective analysis of LMD files on 1210 studies for 935 patients (average age, 39.8±14.7 years old) including 84 children under 18 years old. The study was performed repeatedly for some patients. Phenotyping of CD56+CD19+dim cells was performed using a panel of antibodies to B cell, T/NK cell antigens. The occurrence of blood samples containing CD56+CD19+dim was 1.2%, with a relative content of 2.1±1.9% among total lymphocyte population (0.8±0.6% of leukocytes). Long-term persistence of the subpopulation was noted in the patients throughout the entire observation period. The comparison of specific marker expression by NK CD56+CD19+dim, and CD56+CD19- cells revealed high expression of CD2, CD57, reduced expression density of CD7, CD16, CD38. The phenotype of the studied NK cell subpopulation was as follows: CD56+dimCD19+dimCD2+brightCD7+dimCD11c+CD16+dimCD38+dimCD45RA+CD57+CD94+dimNKG2D+CD3-CD4-CD5-CD20-CD21-CD25-CD45R0-CD62L-CD79b-CD117-, with variable expression of CD8 and HLA-DR. The phenotype is consistent with activated terminally differentiated adaptive NK associated with cytomegalovirus infection. The individuals with CD56+CD19+dim had a history of CMV-infection and reactivation of chronic EBV-infection at the time of the study. A probable cause of CD19 coexpression may be trogocytosis of B cell membrane fragments by natural killer cells during active EBV-infection. CD56+CD19+dim lymphocytes can reach noticeable values thus altering the results of studies performed by flow cytometry. The errors are most likely to occur upon assessing the minimal residual disease levels in acute B cell leukemias. The minor CD56+CD19+dimNK subpopulation may be detected in routine immunological analysis. Its functional features and association with certain disorders require further studies.

PD-1 Blockade Combined with R-CHOP in Patients with Newly Diagnosed EBV-Positive Diffuse Large B-Cell Lymphoma: A Retrospective Analysis

Background Epstein-Barr virus (EBV) -positive diffuse large B cell lymphoma (EBV+ DLBCL) is a relatively rare aggressive B-cell lymphoma. Most patients present with advanced disease, high incidence of extranodal involvement of 30% to 80% (Keane, Eur J Haematol, 2019), as well as poor prognosis when treated with immunochemotherapy. However, no consensus on standard treatment has been established by now. It was found that the expression of programmed death ligand-1 (PD-L1) was increased in EBV+ DLBCL (Kim, Cancer Res Treat, 2019). In addition, chronic EBV infection results in T-cell exhaustion that can be reversed by PD-1 blockade. Compared with PD-L1-positive EBV-negative DLBCL cell lines, the application of PD-1 blockade in PD-L1-positive EBV+ DLBCL cell lines can significantly restore T cell proliferation and activity(Quan, PLoS One, 2015), suggesting that targeting the PD-L1/PD-1 pathway represents a potential therapy for EBV+ DLBCL. Therefore, we retrospectively analyzed the data of nineteen patients with EBV+ DLBCL who were treated with PD-1 blockade combined with RCHOP in our center, hoping to provide reference for subsequent research. Methods Eligible pts had treatment-naive histologically confirmed diagnosis of EBV+ DLBCL. Additional eligibility criteria included at least one measurable lesion, acceptable haptic and hematological function. Pts with primary CNS lymphoma, primary mediastinal large B-cell lymphoma, secondary CNS involvement were excluded. Additional exclusion criteria included history of immune system disease, uncontrolled infection. Once deemed eligible, pts will receive six cycles of RCHOP combined with PD-1 blockade(21 days per cycle) as induction therapy and subsequently maintenance with PD-1 blockade for 1 year(21 days per cycle). End of treatment(EOT) responses were assessed by PET-CT according to the Lugano Classification 2014. The primary objective was to assess the overall response rate(ORR) at EOT. Results Between May 1, 2019, and Sep 1, 2022, 19 pts were enrolled. Pts' baseline demographics, clinical and pathological characteristics are shown in Table 1. The median age was 64 years old without significant gender difference. The majority of pts were intermediate- to high-risk by IPI. EBV DNA can be detected in blood serum at diagnosis in more than half of all patients. No patient was diagnosed with double expressor lymphoma among 18 specimens that were detected c-Myc and BCL-2 with immumohistochemical staining. EBV-encoded RNA (EBER) was positive(>80% of the tumor cells) in all specimens. PD-L1 positivity(>10% of the tumor cells) or CD30 positivity(>5% of the tumor cells) was also quite common. Besides, no double hit lymphoma was confirmed in patient with Fluorescence situ hybridization test(n=13). Treatment course and efficacy evaluation of all pts are summarized in Figure 1. All patients enrolled in this study had completed 6 cycles of induction therapy. Totally, the ORR at EOT was 94.7%, and all achieved CMR. The ORR of patients with PD-L1 positivity(n=10) was numerically higher than that of patients with negative PD-L1 expression (n=5) (100% vs 80%). EBV DNA detected in blood serum of thirteen patients have decreased to undetectable level after induction therapy. The immune-related adverse events (irAEs) were emphatically analyzed. One patient had recurrent fever after every dose of PD-1 blockade without obvious evidence of infection. Another patient developed grade 3 diarrhea since the second cycle of PD-1 blockade maintenance. Colonoscopy showed mucosal swelling and multiple superficial ulcers in the whole colon, which indicated immune colonitis. Maintenance therapy was therefore discontinued. No irAE was observed in the rest of patients. With a median follow-up time of 13.1m (7.2-38.5m), the median PFS and OS were not reached. Both of 1 year OS and PFS was 89.5%. Overall, 2 pts experienced disease progression (1 at EOT, 1 at 2 months after ASCT), 2 pts died (1 due to progressive disease, 1 due to COVID-19 infection ). Conclusions PD-1 blockade combined with R-CHOP as induction therapy in patients with newly diagnosed EBV+ DLBCL has demonstrated satisfactory response and limited irAEs. Longer follow-up is still warranted to confirm the long-term benefit, especially for patients with maintenance of PD-1 blockade. Still, we believe this therapeutic strategy can be regarded as a successful attempt at improving prognosis for pts with EBV+ DLBCL.

Characteristics of self-regulation of the epidemic process of infection caused by the Epstein-Barr virus (Herpesviridae: Lymphocryptovirus, HHV-4).

Among the available scientific literature, there are no publications addressing processes of self-regulation in the parasite-host population systems with reference to chronic infections, including the infection caused by the Epstein-Barr virus (EBV infection). The aim of the study is to assess manifestations of the epidemic process of chronic EBV infection through the lens of the basic tenets of the theory of self-regulation of parasitic systems. The study was performed using data from scientific publications selected from such database sources as Scopus, Web of Science, Cochrane Library, PubMed, CyberLeninka, RSCI, etc. The list of analyzed publications included published articles of the authors of this study, reporting the results of the retrospective epidemiological analysis of the incidence of infectious mononucleosis in Russia in general and in Moscow in particular, as well as the results of the laboratory tests regarding the detection frequency of specific antibodies to EBV proteins. The chronic course of EBV infection promotes a close long-term interaction between the pathogen and the host. The genetic variability of the pathogen and the functions of specific and nonspecific human immune defense systems play a key role in the interaction between two heterogeneous populations and underlie their phasal self-transformation. A variety of social and natural factors (adverse chemical, physical, biological, climatic impacts, etc.) trigger the reactivation of chronic EBV infection, thus providing the continuous existence of additional sources of infection in the host population. The analysis of the manifestations of chronic EBV infection in the context of the theory of self-regulation of parasitic systems promotes the understanding of the factors underlying the unevenness of its epidemic process. The obtained data can be adjusted for other infections having similar transmission mechanisms and virus life cycles (including other herpes infections) to map out strategies to control the epidemic process of chronic infections spread by aerosol transmission of the pathogen.

Epigenetic activation of the TUSC3 gene as a potential therapy for XMEN disease

"X-linked MAGT1 deficiency with increased susceptibility to EBV infection and N-linked glycosylation defect" (XMEN) disease is a rare combined immunodeficiency (CID) caused by loss of function mutations in the magnesium transporter 1 (MAGT1) gene. MAGT1 deficiency impairs magnesium transport and the N-glycosylation of a panel of proteins, which subsequently abolishes the expression of key immune receptors such as Natural-Killer Group 2, member D (NKG2D). These effects induce immune system abnormalities, chronic EBV infection and neoplasia. Recent research shows that MAGT1 and tumor candidate suppressor 3 (TUSC3) share high sequence and functional similarity. We sought to investigate the feasibility of activating TUSC3 expression to provide a potential therapeutic strategy for XMEN disease. The expression profiles of MAGT1 and TUSC3 were analyzed using multiple databases, RT‒PCR, and western blot. The effects of decitabine and panobinostat on the regulation of TUSC3 expression were explored in both MAGT1 knock-out (KO)/patient-derived lymphocytes and MAGT1 KO hepatocytes. Although TUSC3 is widely expressed, it is undetectable specifically in the immune system and liver, consistent with the main diseased tissues in XMEN patients. CRISPR/Cas9-mediated KO of MAGT1 in the NKL cell line successfully mimicked the phenotypes of XMEN patient-derived lymphocytes, and exogenous expression of TUSC3 rescued the deficiencies in KO NKL cells. Using this in vitro model, we identified two epigenetic drugs, decitabine and panobinostat, by screening. Combination treatment using these two drugs significantly upregulated TUSC3 expression and rescued the immune and liver abnormalities. Epigenetic activation of TUSC3 expression constitutes an effective therapeutic strategy for XMEN disease.

MicroRNA Gene Signature for Predicting Mechanisms in Nasopharyngeal Carcinoma: A Case Study on the Potential Application of Circulating Biomarkers.

Nasopharyngeal Carcinoma (NPC) is an upper respiratory tract cancer prevalent in Southeast Asia and related to chronic EBV infection. microRNAs (miRNAs) regulate gene expression implicated in NPC's carcinogenesis. However, this circulating RNA molecule's role and clinical utility remain unknown. Therefore, this study examined the circulation of miRNAs and their association with clinical data. 160 plasma samples of NPC and 80 non-tumor samples were extracted to evaluate and validate the gene expressions. Quantification expression was performed using relative quantification of qPCR analysis level expression methods. The intrinsic cellular roles involving biological signaling in NPC's oncogenesis using Ingenuity Pathways Analysis (IPA) were also used. The results of the quantification significance profiling of NPC samples revealed decreased miR- 29c-3p (fold change 1.16; p<0.05) and increased 195-5p expression (fold change 1.157; p<0.05). Furthermore, the validation of hsa-miR-29c-3p expression on plasma NPC with known tumor vs. non-tumor and significant changes was also performed using a fold change of 4.45 (medians of 31.45 ± 1.868 and 24.96 ± 1.872, respectively; p<0.0005). miR-29c had a 2.14 fold change correlated with T primary status with a median of 31.99±1.319 and 31.35±2.412, respectively (p<0.05). Stage status with fold change 1.99 also had median levels of 31.98±1.105 and 31.21 ± 2.355, respectively (p-value <0.05). Furthermore, the node's status for the lower expression of miR-29c with fold change 1.17 had median levels of 32.78 ± 2.221 and 31.33 ± 1.689, respectively (p-value of 0.7). Bioinformatics analysis established the roles and functions of miR-29 in NPC progression, cell death and survival, cellular development, cellular function, and cell maintenance by inhibiting COL4A, PI3K, VEGFA, JUN, and CDK6. Overall, we conclude that decreased miR-29c expression is associated with poor clinical status and might inhibit NPC's five target genes.

Incorporating Immunotherapy in the Management of Gastric Cancer: Molecular and Clinical Implications.

Simple SummaryGastric cancer is one of the most common malignant tumours worldwide, with the fifth and third highest morbidity and mortality, respectively, of all cancers. Survival is limited, as most of the patients are diagnosed at an advanced stage, and are not suitable for surgery with a curative intent. Chemotherapy has only modestly improved patients’ outcomes and is mainly given with a palliative intent. Immunotherapy has improved overall survival of patients with gastric cancer, and has thus become a new standard of care in clinic. In this review we discuss the strong molecular rationale for the administration of immunotherapy in this disease and analyse the clinical data supporting its use.Gastric cancer has a median survival of 11 months, and this poor prognosis has not improved over the last 30 years. Recent pre-clinical data suggest that there is high tumour-related neoantigen expression in gastric cancer cells, suggesting that a clinical strategy that enhances the host’s immune system against cancer cells may be a successful approach to improve clinical outcomes. Additionally, there has been an increasing amount of translational evidence highlighting the relevance of PD-L1 expression in gastric cancer cells, indicating that PD-1/PD-L1 inhibitors may be useful. Several molecular subgroups of gastric cancer have been identified to respond with excellent outcomes to immunotherapy, including microsatellite instable tumours, tumours bearing a high tumour mutational burden, and tumours related to a chronic EBV infection. In gastric cancer, immunotherapy has produced durable responses in chemo-refractory patients; however, most recently there has been a lot of enthusiasm as several large-scale clinical trials highlight the improved survival noted from the incorporation of immunotherapy in the first line setting for advanced gastric cancer. Our review aims to discuss current pre-clinical and clinical data supporting the innovative role of immunotherapy in gastric cancer.

Epstein-Barr Virus and Malaria Interactions: Immunology Perspective

Epstein-Barr Virus can cause various diseases, from acute inflammatory diseases such as fatal or chronic EBV infection, infectious mononucleosis as well as lymphoid and epithelial cancer, various autoimmune diseases, and also could interact with malaria. As EBV infects 95% of the world population, and more than 30% are infected with the protozoan parasite, with more than 500,000 deaths due to malaria cases. It is important to understand how EBV dysregulates the immune system, especially when the virus is interacting with other pathogens such as malaria parasites, causing more severe conditions in certain people like Burkitt Lymphoma. This review will be informative about the mechanism of how EBV interacts with malaria parasites and how it affects the immune system. Knowledge of various cytokines triggering the immune system which may provide links to control/minimize malaria disease severity.

Abstract 6127: Single-cell spatial transcriptome sequencing characterizes CD70+ immune-resilient nasopharyngeal carcinoma cells as a vital mediator of Treg-induced suppression

Abstract Background: The microenvironment of nasopharyngeal carcinoma (NPC) has an intense and heterogenous immune infiltration, due to chronic EBV infection and locoregional lymphoid structures. Phase II clinical trials have shown that the response rate of NPC patients to PD-1 inhibitors is only 20%, indicating the current immunotherapies are not optimal due to insufficient understanding of the tumor-immune interaction. In this study, we have applied an integrated transcriptome sequencing analysis to decipher immune dynamics, and have characterized a new target that can inhibit Treg-mediated suppression in NPC, and provided critical translational evidence to the precision immunotherapy. Methods: We established an integrated NPC single-cell sequencing cohort containing 189,750 T cells from 50 patient samples. Spatial transcriptome sequencing was performed on frozen samples from eight NPC patients. The whole blood was collected from healthy donors and PBMCs were isolated using density gradient centrifugation. T cells were magnetically isolated from PBMCs. The humanized mouse model was established by tail-vein injection of CD3/CD28 activated PBMCs. Results: Single-cell sequencing revealed the enrichment of suppressive Tregs in the NPC microenvironment. We found that the C666-1 cell line facilitated Treg differentiation and activation. Spatial sequencing identified CD70-CD27 interaction was upregulated between NPC cells and suppressive Tregs, indicating its role in regulating Treg function. CD70 was predominantly expressed on NPC cells and correlated to worse prognosis. CD70-knockout (KO) in C666-1 inhibited Treg differentiation. Decreased Treg activities further enhanced the cytotoxicity in CD8+ effector T cells, leading to higher tumor apoptosis. Cusatuzumab also generated a comparable inhibitory effect in Treg-mediated suppression. In the humanized mouse model, CD70-KO and inhibition shrank the size of subcutaneous tumors by enhancing T cell immunity. Conclusion: The enrichment and activation Tregs makes the NPC microenvironment highly immunosuppressive, which might jeopardize the clinical efficacy of anti-PD1 therapies in NPC patients. In this study, we revealed that CD70+ NPC cells were highly resilient to immune surveillance since they were capable of modulating Treg differentiation and activation via the CD70-CD27 interaction to counteract CD8+ T cells. Blocking CD70 has been shown to effectively induce tumor apoptosis by alleviating Treg-mediated suppression. The therapeutic efficacy and safety of the anti-CD70 therapy were tested in the humanized mouse model. These findings suggested that the anti-CD70 therapy exhibited a promising efficacy to kill NPC cells by deteriorating the immunosuppressive microenvironment, and it might synergistically enhance the efficacy of anti-PD1 therapies. Citation Format: Lanqi Gong, Jie Luo, Yuma Yang, Yu Zhang, Xin-Yuan Guan. Single-cell spatial transcriptome sequencing characterizes CD70+ immune-resilient nasopharyngeal carcinoma cells as a vital mediator of Treg-induced suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6127.

Further Delineation of the Spectrum of XMEN Disease in Six Chinese Pediatric Patients

X-linked MAGT1 deficiency with increased susceptibility to EBV-infection and N-linked glycosylation defect (XMEN) disease is a primary immunodeficiency caused by loss-of-function variants in the MAGT1 gene. Only two patients from one family have been diagnosed with XMEN in China. In this study, we retrospectively analyzed the genetic, clinical, and immunological characteristics of six pediatric patients in a Chinese cohort. Medical records were retrieved, immunological phenotypes were assessed, and infectious microbes in patients were detected. Six male patients (mean age, 6.3 years) from five unrelated families were genetically diagnosed as XMEN. Five patients presented with a major complaint of elevated liver enzymes, while one patient was referred for recurrent fever, cough and skin rash. Five patients developed EBV viremia, and one patient developed non-Hodgkin’s lymphoma. Histopathological findings from liver biopsy tissues showed variable hepatic steatosis, fibrosis, inflammatory infiltration, and glycogenosis. Immune phenotypes included CD4 T-cell lymphopenia, elevated B cells, inverted CD4/CD8 ratios, and elevated αβDNTs. No pathogenic microbes other than EBV were identified in these patients. This study reports the clinical and molecular features of Chinese patients with XMEN. For patients with transaminase elevation, chronic EBV infection and EBV-associated lymphoproliferative disease, the possibility of XMEN should be considered in addition to isolated liver diseases.

ANAMNESTIC, CLINICAL AND LABORATORY DATA ANALYSIS OF PATIENTS FOR DRUG HYPERSENSITIVITY REACTIONS

The current development of the pharmaceutical industry in the synthesis of new chemical compounds, standardized treatment protocols, and disease prevention can lead to a progressive increase in drug hypersensitivity reactions, which often have serious consequences for human health. Increasing evidence of involvement of infections, including Herpesviridae viruses, in the development of drug hypersensitivity reactions is known.The method of flow cytometry can be used, in particular, the basophil activation test to diagnose drug hypersensitivity reactions. The anamnestic, clinical, andlaboratory data of 368 people were analyzed for the selection of patients at risk of drug hypersensitivity for the basophil degranulation test execution. It was found that among patients hypersensitivity reactions were most often detected to antibiotics (50.0%), radiopaque substances (27.7%), perioperative drugs, local anesthetics - 13.6% each. Clinical manifestations of these reactions were urticaria with angioneurotic edema (40.6%), urticaria (28.1%), anaphylaxis (21.9%), obstructive bronchitis chenges (9.37%). According to anamnestic and clinical-laboratory data, patients with a high risk of drug hypersensitivity reactions revealed frequent manifestations of herpesvirus infection HSV1 (34.4%), active chronic persistence of EBV (59.4%), accompanied by manifestations of EBV-associated secondary immune disorders and prevalence of chronic EBV infection in all patients.

ІМУНОЗАЛЕЖНІ МЕХАНІЗМИ ТА МОЛЕКУЛЯРНО-ГЕНЕТИЧНІ ОСОБЛИВОСТІ У ХВОРИХ НА СИСТЕМНІ ХВОРОБИ СПОЛУЧНОЇ ТКАНИНИ З КРІОГЛОБУЛІНЕМІЧНИМ СИНДРОМОМ

Introduction: Cryoglobulinemic syndrome (CGS) is an immune-related process caused by cryoglobulins composition in the blood in small or medium vessels. Most frequently, CGS is triggered by lymphotropic viruses, immune-related and oncological diseases. Objectives: Studying the immune-related mechanisms, molecular and genetic characteristics of patients with systemic autoimmune diseases (SAD) against the cryoglobulinemic syndrome. Methods: Among 380 patients with SAD, in 94 (57.6%) progressing chronic EBV-infection was diagnosed, and 22.1% of patients were diagnosed with progressing chronic HSV 1/2-infection based on DNA virus identification through the polymerase chain reaction (PCR) in three biological media (blood, saliva, mucus membrane scraping). Results: Analysis of the cryoglobulins in such patients showed that CGS was diagnosed in 118 (31.1%) patients with the mean concentration of CG1.68±0.33 g/l at a rate of 0.48 ±0.10g/l. The patients with the systemic connective tissue diseases with CGS demonstrated statistically lower miR-146а expression which resulted in the abnormal production of pro-inflammatory cytokines, the highest TLR9 expression on monocytes, slightly lower on lymphocytes, and the lowest on granulocytes; the increase in the relative amount of cytolytic T-lymphocytes, IL2 receptor lymphocytes, activated CD HLA DR+-lymphocytes against the reduction of NK-cells and regulatory suppressor CD4+/25+-cells was observed. The idiopathic and initiated oxidative monocyte capacity in CGS patients distinctly tended to increase, as compared to patients without CGS and normal individuals. Conclusions. Cryoglobulins may act as the so-called bridge between viral infections and the autoimmune processes. CGS was diagnosed in 31.1% of patients. Despite a substantial number of studies dedicated to the cryoglobulinemic syndrome, the peculiarities of the immune reaction of such patients need further research, since they create the risks of secondary vasculitis against SAD

Gene expression analysis in EBV-infected ataxia-telangiectasia cell lines by RNA-sequencing reveals protein synthesis defect and immune abnormalities

BackgroundEpstein–Barr virus (EBV) targets B-cells where it establishes a latent infection. EBV can transform B-cells in vitro and is recognized as an oncogenic virus, especially in the setting of immune compromise. Indeed, immunodeficient patients may fail to control chronic EBV infection, leading to the development EBV-driven lymphoid malignancies. Ataxia telangiectasia (AT) is a primary immune deficiency caused by mutations in the ATM gene, involved in the repair of double-strand breaks. Patients with AT are at high risk of developing cancers, mostly B-cell lymphoid malignancies, most of which being EBV-related. Aside from immune deficiency secondary to AT, loss of ATM function could also hinder the control of the virus within B-cells, favoring lymphomagenesis in AT patients.ResultsWe used RNA sequencing on lymphoblastoid cell lines derived from patients with AT and healthy donors to analyze and compare both cellular and viral gene expression. We found numerous deregulated signaling pathways involving transcription, translation, oncogenesis and immune regulation. Specifically, the translational defect was confirmed in vitro, suggesting that the pathogenesis of AT may also involve a ribosomal defect. Concomitant analysis of viral gene expression did not reveal significant differential gene expression, however, analysis of EBV interactome suggests that the viral latency genes EBNA-3A, EBNA-3C and LMP1 may be disrupted in LCL from AT patients.ConclusionOur data support the notion that ATM deficiency deregulates cellular gene expression possibly disrupting interactions with EBV latent genes, promoting the oncogenic potential of the virus. These preliminary findings provide a new step towards the understanding of EBV regulation and of AT pathogenesis.

Efficacy and Relapse Prediction Model of Allogeneic Peripheral Blood Stem Cell Transplantation in Adult Acute Leukemia

To observe the clinical efficacy of allogeneic peripheral blood stem cell transplantation（allo-HSCT） on the treatment of adult acute leukemia patients, moreover, to establish and evaluate a Logistic model to predict the risk of relapse in adult acute leukemia patients after allo-HSCT. The clinical data of 145 adult acute leukemia patients treated by peripheral blood stem cell transplantation in the First Affiliated Hospital of Xi'an Jiaotong University from January 2010 to December 2019 was enrolled and analyzed retrospectively. Complications and survival of patients were observed. The relationship between patients' age, diagnosis, leukocyte count at onset, risk stratification, time of diagnosis to transplantation, HCT-CI, minimal residual disease pre-transplantation, donor-recipient sex relationship, HLA match degree, prophylaxis of graft versus host disease(GVHD), donor age, number of transfused mononuclear cells, CD34 positive cells, engraftment time, acute and chronic GVHD, CMV, EBV infection, and hemorrhagic cystitis and recurrence after transplantation were analyzed by logistic regression. Relapse prediction model was established and evaluated according to the results. Among 145 acute leukemia patients, 81 with acute myeloid leukemia, 64 with acute lymphocytic leukemia, 18 with EBV infection, 2 with post-transplant lymphoproliferative disorder(PTLD), 85 with CMV, 26 with hemorrhagic cystitis, 65 patients developed acute GVHD, 51 patients developed chronic GVHD and 45 patients relapsed. The overall survival （OS） rates in one and three years were 86.4% and 61.8%, and the progress-free survival （PFS） rates in one and three years were 67.5% and 62.4%, respectively. There were significant differences in OS and PFS between relapsed and non-relapsed patients, as well as AML and ALL patients. Univariate analysis revealed that patient's age, risk stratification, time to transplantation, HCT-CI index, ATG based GVHD prophylaxis, minimal residual disease pre-transplantation, GVHD prophylaxis, and acute and chronic GVHD were associated with the relapse of disease, multivariate logistic regression analysis showed that pre-transplantation minimal residual disease showed positively correlation with relapse of the disease, while chronic GVHD showed negatively correlation. The relapse rate of adult acute leukemia patients treated with allo-HSCT in our hospital is 31.0%, and OS of AML patients is better than ALL patients'. OS of relapsed patients is significantly lower than non-relapsed patients'. Pre-transplantation minimal residual disease is a risk factor of relapse. The risk of relapse is reduced in patients with chronic GVHD.

АНАЛІЗ ВЗАЄМОЗВ’ЯЗКУ РІВНІВ КІНЦЕВИХ ПРОДУКТІВ ГЛІКАЦІЇ З ЦИТОКІНОВИМ ПРОФІЛЕМ У ПАЦІЄНТІВ З АЛЕРГІЧНИМИ ХВОРОБАМИ НА ТЛІ ХРОНІЧНОЇ ЕПШТЕЙНА-БАРР ВІРУСНОЇ ІНФЕКЦІЇ

ANALYSIS OF THE CORRELATION BETWEEN ADVANCED GLYCATION END-PRODUCTS LEVELS AND THE CYTOKINE PROFILE IN PATIENTS WITH ALLERGIC DISEASES ASSOCIATED WITH CHRONIC EPSTEIN-BARR VIRAL INFECTION S. O. Zubchenko Danylo Halytsky Lviv National Medical University, Lviv, Ukraine Abstract. Advanced glycation end-products (AGEs) serve as densitometric markers of oxidative stress and inflammatory processes in many diseases and their complications. Objective: analysis of the correlation between AGEs and the cytokine profile in patients with allergic diseases associated with chronic Epstein-Barr viral infection (EBV-infection) in the active and latent phases. Object and methods. Determination and comparative analysis of AGEs levels and cytokines IL-17, IL-33, TNF-α in 66 patients aged (32.4 ± 7.5) years with allergic diseases associated with chronic EBV-infection in the active and latent phases and in EBV-seronegative patients with allergic diseases. Results. When determining AGEs by fluorescence spectroscopy in three groups of patients with allergic diseases, there was a reliably smaller number of AGEs than in healthy individuals. Presumably 12.3 % higher levels of IL-33 (p = 0.041) was determined in patients with active phase of EBV-infection compared to EBV-seronegative individuals, and in patients with latent phase of EBV-infection, the level of IL-17 was 3.54 times higher compared with EBVseronegative individuals (p = 0.011). It was determined that there is a reliable (p = 0.049) weak negative (r = -0.364) correlation between AGEs level and IL-33 in patients with allergic diseases associated with chronic EBV-infection in the active phase. It was investigated that no statistical difference between the levels of the proinflammatory cytokine TNF-α in the three groups was found. In patients with allergic diseases associated with chronic EBV-infection in the latent phase, a reliable (p = 0,032) weak negative (r = -0,364) correlation between AGEs and IL-17 was determined. Conclusions. Patients with allergic diseases compared to healthy individuals have lower levels of AGEs. AGEs play a significant pathogenetic role in the induction of allergic inflammatory process. AGEs accompanied by increased synthesis of IL-33 and IL-17 was more pronounced in patients with active phase EBV-infection, which was confirmed by clinical data. Key words: аdvanced glycation end-products, allergic diseases, chronic Epstein-Barr viral infection, cytokines. S. Zubchenko PhD, Associate Professor, Department of Clinical Immunology and Allergology Danylo Halytskyi Lviv National Medical University Pekarska street, 69, Lviv, Ukraine, 79010. Svitlanazu@gmail.com Аsthma and Allergy, 2021, 3, P. 30–35.

THE EFFECTIVENESS OF THERAPY BY CRYOPRESERVED HUMAN PLASMA IN PATIENTS WITH DEFICIENCY OF MANNOSE BINDING LECTIN SUFFERING FROM HERPES VIRUS INFECTION

The aim: To research the effectiveness of cryopreserved blood plasma replacement therapy in patients with primary mannose binding lectin (MBL) deficiency, suffering from chronic active herpes virus infections. Materials and methods: Patients of the study group (SG) n= 36 additionally received cryopreserved blood plasma therapy Octaplas (Octapharma, Switzerland). Patients of the control group (CG) n=36 received only chemotherapy with Valganciclovir 450 mg 2/day per os for 1-3 months. The diagnosis of active herpes virus infection was established by PCR of blood leukocytes. Statistical analysis of the obtained information was processed by the calculation of the chi-square (χ2) Pearson criterion, the odds ratio and the associated 95% confidence interval (95% CI). Results: The adding cryopreserved blood plasma substitute to standard therapy with valganciclovir for the treatment of chronic active herpes virus infection in patients with total serum MBL deficiency below 50 ng/ml, allowed to get more negative PCR results. The effectiveness of combination therapy was 50% higher in carrier of HHV-6 (χ2=8,533 and р=0,004; Yeats correction 6,533 and significance 0,011; OR=11,667 and 95% CI=1,939-70,180) and 43% in carrier of HHV-7 (χ2=8,846 and р=0,003; Yeats correction 7,165 and significance 0,008; OR=6,375 and 95% CI=1,711-23,758), compared with monotherapy. The close association between deficit MBL compensation and the results of antiviral treatment is also reported. The effect of such treatment in patients with chronic EBV infection was less (27%). Conclusions: We assumed, that virostatic effect of valganciclovir is increased by MBL-mediated clearance of blood serum from viral particles.

Effective Control of Epstein-Barr Virus–Related Hemophagocytic Lymphohistiocytosis With Immunochemotherapy

The familial form of hemophagocytic lymphohistiocytosis (HLH) is a lethal disorder. Although the prognosis for Epstein-Barr virus-associated HLH (EBV-HLH) remains uncertain, numerous reports indicate that it can also be fatal in a substantial proportion of cases. We therefore assessed the potential of immunochemotherapy with a core combination of steroids and etoposide to control EBV-HLH in 17 infants and children who met stringent diagnostic criteria for this reactive disorder of the mononuclear phagocyte system. Treatment of life-threatening emergencies was left to the discretion of participating investigators and typically included either intravenous Ig or cyclosporin A (CSA). Five patients (29%) entered complete remission during the induction phase (1 to 2 months), whereas 10 others (57%) required additional treatment to achieve this status. In 2 cases, immunochemotherapy was ineffective, prompting allogeneic bone marrow transplantation. Severe but reversible myelosuppression was a common finding; adverse late sequelae were limited to epileptic activity in one child and chronic EBV infection in 2 others. Fourteen of the 17 patients treated with immunochemotherapy have maintained their complete responses for 4+ to 39+ months (median, 15+ months), suggesting a low probability of disease recurrence. These results provide a new perspective on EBV-HLH, showing effective control (and perhaps cure) of the majority of EBV-HLH cases without bone marrow transplantation, using steroids and etoposide, with or without immunomodulatory agents.

Epstein-Barr virus-positive diffuse large B-cell lymphoma, not otherwise specified: A diagnostic challenge

Epstein-Barr virus-positive (EBV+) diffuse large B-cell lymphoma (DLBCL), not otherwise specified (NOS) is an entity included in the 2016 World Health Organization classification of lymphoid neoplasms. EBV+DLBCL, NOS, is an aggressive B-cell lymphoma associated with chronic EBV infection and poor prognosis with standard chemotherapeutic approaches. We present the case of a 71-year-old male hospitalized for generalized lymphadenopathy, unexplained weight loss, and intermittent fever with pancytopenia and a very high EBV viral load. An inguinal lymph node was biopsied and reported at two different centers as Classic Hodgkin lymphoma and EBV associated lymphoproliferative, respectively. The patient was subsequently rebiopsied and a cervical lymph node was subjected to detail histopathologic and immunohistochemical evaluation by clonality studies resulting in a final diagnosis of EBV positive DLBCL, NOS. This case highlights the potential diagnostic pitfalls due to the morphologic heterogeneity of this entity and immunohistochemical overlap with Hodgkin lymphoma, T-cell rich large B-cell Lymphoma, and diffuse large B-cell lymphoma (DLBCL, NOS).

EBV-positive diffuse large B-cell lymphoma, not otherwise specified: 2020 update on diagnosis, risk-stratification and management.

Epstein Barr virus-positive (EBV+) diffuse large B-cell lymphoma (DLBCL), not otherwise specified (NOS) is an entity included in the 2016 WHO classification of lymphoid neoplasms. EBV+ DLBCL, NOS, is an aggressive B-cell lymphoma associated with chronic EBV infection, and a poor prognosis with standard chemotherapeutic approaches. The diagnosis is made through a careful pathological evaluation. Detection of EBV-encoded RNA (EBER) is considered standard for diagnosis; however, a clear cutoff for positivity has not been defined. The differential diagnosis includes plasmablastic lymphoma (PBL), DLBCL associated with chronic inflammation and primary effusion lymphoma (PEL), among others. The International Prognostic Index (IPI) and the Oyama score can be used for risk-stratification. The Oyama score includes age >70 years and presence of B symptoms. The expression of CD30 and PD-1/PD-L1 are emerging as potential adverse but targetable biomarkers. Patients with EBV+ DLBCL, NOS, should be staged and managed following similar guidelines than patients with EBV-negative DLBCL. EBV+ DLBCL, NOS, however, might have a worse prognosis than EBV-negative DLBCL in the era of chemoimmunotherapy. There is an opportunity to study and develop targeted therapy in the management of patients with EBV+ DLBCL, NOS.