Chronic Herpesvirus Infection Research Articles

Mice microglia cytokine profile changes under the influence of HSV-1

Introduction. Today, prevalence of neurodegenerative diseases increases. In recent years, more studies have revealed a new knowledge about the role of microglia in the development of these diseases. Animal experiments showed that peripheral inflammation causes activation of microglia in brain. All this points to the essential role of the cells in the development of neurodegeneration. Under the influence of various factors, microglia can change the phenotype and participate in both repair and damage to brain cells. Chronic herpesvirus infection caused by HSV-1 is another known factor in the development of neurodegenerative pathology. However, the exact pathogenetic mechanisms are still unknown, nevertheless, studying the virus effect on microglia has great potential. The goal of our study in this connection was to assess the effect of HSV-1 on microglia polarization in mouse strain with normal susceptibility to this virus and in strain which is more resistant to the action of HSV-1. For this purpose, changes in the cytokine profile were detected. A comparison of interstrain differences in the expression of cytokine genes was also compared in control groups. Materials and methods. The study involved infecting C57BL/6 and BALB/c mice with herpes simplex virus type 1, an isolation of microglia was based on separation steps using a discontinuous gradient density, the cytokine profile was assessed by gene expression levels using a real-time reverse transcription PCR. To calculate the relative fold gene expression of samples the 2–ΔΔCt method was used. Statistical significance was determined using the Mann–Whitney U-test. Results. There were found no interstrain differences in cytokine gene expression in control groups of different mouse strains. At the same time, gene expression differed in the experimental groups: in BALB/c mice, the expression of genes for both pro-inflammatory and anti-inflammatory cytokines increased; in C57BL/6 mice, a slight increase in the expression of IL-1β genes was observed. Conclusion. The data indicate the formation of different microglial phenotypes after HSV-1 infection in different mouse strains. Apparently, in BALB/c mice there is a switch from the pro-inflammatory M1 phenotype of microglia to the anti-inflammatory M2 phenotype, while in C57BL/6 mice the attenuation of the infectious process occurs through a return to the original M0 phenotype.

Variability of changes in proand antiinflammatory cytokines due to IFNα and IFNγ deficiency in patients with post-covid syndrome associated with activation of chronic herpes viral infections

Post-COVID syndrome (PCS) is characterized by long-term complications and conditions accompanied by neuroimmunoinflammation, and includes chronic fatigue syndrome (CFS) and cognitive disorders (CD), which are often associated with activation of chronic herpesvirus infections (HVI). Timely detection of symptoms and immunodiagnosis of PCS are a priority and are of undoubted interest. Objective: to clarify the levels of serum proand anti-inflammatory cytokines, alpha and gamma interferons in patients with post-COVID syndrome associated with confirmed activation of chronic herpesvirus infections. Patients (n = 60) aged from 18 to 65 years with complaints of manifestations of PCS associated with HVI were studied — the study group (SG). A survey was conducted using a modified scale-questionnaire to assess the severity of PCS symptoms in points from 0 to 4, real-time PCR of HVI (EBV, HSV1/2, VCH6, VCH8, CMV) in saliva and scrapings from the tonsils, determination of the level of IFNα and IFNγ, pro- (TNFα, IL-18, IL-1β, IL-6, IL-17A, IL-8) and anti-inflammatory (IL-4 and IL-10) cytokines in blood serum. Comparison group (CG) — 60 apparently healthy individuals. SG patients with mixed HVI with EBV dominance noted the most pronounced and persistent clinical manifestations of PCS, among which the leading place was occupied by longterm sensations of CFS and CD. A persistent multisystem inflammatory response was identified, confirmed by elevated levels of IL-6 and IL-17A, which caused severe PCS. In post-COVID period, hyperproduction of IL-1β was detected, which was accompanied by clinical manifestations of persistent neuroimmunoinflammation. At the same time, the identified deficiency of IFNα, IFNγ and dysregulatory disorders in the antiviral defense of the immune system in patients with PCS contributed to the activation of HVI. Data on the imbalance of proand anti-inflammatory cytokines in the SG were obtained, which confirms the presence of a persistent multisystem inflammatory reaction with dominance of persistent neuroimmunoinflammation, which causes severe PCS. An imbalance of the cytokine system with IFNα and IFNγ deficiency, associated with the activation of chronic HVI with EBV dominance in the post-COVID period, contributes to the development of neuroimmunoinflammation, which is accompanied by the leading clinical signs of PCS: CFS and CD.

Efficacy of differentiated targeted interferon and immunomodulatory therapy focused on pathological immunophenotypes in patients with atypical chronic active herpesvirus infections

Objective: In a prospective cohort study to evaluate the clinical and immunological efficacy of differentiated targeted interferon and immunomodulatory therapies focused on identified pathological immunophenotropes and associated clinical manifestations in immunocompromised patients suffering from atypical chronic active herpes viral infections (ACA-HVI).Materials and methods: 335 patients suffering from mixed-AHA-HVI were examined. The study complex included: methods for detecting herpesviruses: serodiagnosis, PCR-RV; immunological methods: a research of subpopulation of blood lymphocytes (method of a flow cytometry), determination of the spontaneous and induced products of IFNα and IFNγ, levels of serum cytokines and immunoglobulins (IL-1β, IL-6, IL-17, TNFα and IFNα and IFNγ, Ig A, M, G) by ELISA. The study was approved by the ethics board and informed consent was obtained from all patients.Results: Integral formulas of disorders in the antiviral immune defense system were created, which made it possible to isolate 3 pathological immunophenotypes (PIF): PIF1: NG↓+ind.IFNα/IFNγ↓+CTL↓+EKK↓+IgM↑+hypecytokinemiya (IL-1β↑+IL-6↑+TNFα↑); PIF2: NG↓+ ind. IFNα/IFNγ↓ + EKK↓+ IgG↓+hypercytokinemiya (IL-1β↑+IL-6↑+TNFα↑) and PIF3: NG↓+ind.IFNα/IFNγ↓+hypercytokinemiya (IL-1β↑+IL- 6↑+TNFα↑). Taking into account the identified disorders, a program of targeted interferon and immunomodulatory therapy was developed for each PIF: local and systemic IFN therapy + hexapeptide was developed for PIF1; for PIF2- local and systemic IFN therapy + glucosaminimuramyldipeptide; for PIF3-local and systemic IFN therapy.Conclusions: High clinical efficacy was demonstrated in 100% of patients with three groups of ACA-HVI. Immunological effectiveness of targeted interferon and immunomodulatory therapy programs: 89.5% for PIF1; for PIF2-57.6% and for PIF3- 37.5% of cases.

Interferon system in typical chronic and atypically occurring chronic active herpes virus infections

Objective: to study the characteristics of the functioning of the IFN system, the presence of autoantibodies to INFα in patients suffering from atypical chronic active herpesvirus infections (ACA-HVI) in comparison with patients with a typical course of chronic herpesvirus infections (CHVI).Materials and methods: under our supervision were 485 patients of both sexes aged 23 to 70 years, suffering from chronic herpes virus infections, of which 335 patients suffered from AHA-HVI and 150 people suffered from CHVI. The comparison group was 250 conditionally healthy individuals (CG). The complex of the study included methods for detecting herpesviruses: serodiagnostics, PCR-RT. The IFN system (spontaneous and induced production, serum concentration) was tested by ELISA. The study was approved by the ethics board and informed consent was obtained from all patients.Results: the incidence of various mono-mixed herpesvirus infections in patients with ACA-HVI (mono – 26,6% and mixed – 73.4%) and CHVI (mono – 23.1% and mixed – 76.9%) was determined, with EBV dominance in patients of both groups. Serum IFNα deficiency was detected in 100% of cases in both groups, and IFNα in 67% in ACA-HVI and 57% in CHVI. At the same time, significant differences were found between the ACA-HVI and HGVI groups in the level of IFNα reduction: 10 and 5 times, respectively, and for IFNγ – 2.0 and 2.6 times, respectively. The induced IFNα production decreased by 89.1% in ACA-HVI and 47.2% in CHVI. A decrease in induced IFNγ production is characteristic of 50% of patients in both groups. At the same time, the level of induced production of IFN α in patients with ACA-HVI was 9 times lower than in the control group and 4.75 times lower than in the group of patients with CHVI. And the level of induced IFNγ production was 2 times lower compared to CHVI and the control group.Conclusions: when assessing the state of the IFN system in patients with various chronic herpes virus infections, significant differences were revealed. Thus, the most pronounced manifestations of interferonopathy, consisting in a significant decrease in serum IFNα and IFNγ and defects in induced IFN production of both types, are observed statistically significantly more often in the group of patients with an atypical course of the disease than in the group of patients with a typical course of CHVI. The most pronounced disorders in the IFN system and the lack of recovery of IFNα and IFNγ levels in the interrelational period cause atypicity of the course and active viral replication in patients with ACA-HVI.

PATHOGENETIC SIGNIFICANCE OF THE CHARACTER OF WOMAN’S UROGENITAL INFECTION IN THE DEVELOPMENT OF ADENOMYOSIS

Adenomyosis is one of the important problems of modern gynecology, which leads to impaired reproductive function, reduced work capacity and quality of life of women. The prevalence of endometriosis in the female population ranges from 20-55% in patients of reproductive age. An important role in the occurrence and development of adenomyosis is played by genetic predisposition, hormonal changes and the influence of immunological factors. Of great interest is a new concept regarding the significant role of bacterial contamination of the genital tract in patients with adenomyosis. The study of the influence of microorganisms on the mechanisms of activation of the inflammatory reaction associated with the progression of endometrioid lesions is relevant and requires the search for new approaches to diagnosis and treatment of the disease.
 The purpose of the work is to study the influence of the woman’s urogenital infection in the development of adenomyosis.
 Materials and methods. 85 patients with 1st and 2nd degree adenomyosis and 30 healthy fertile women were examined. The average age of the examined was 27.3±1.6 years. A comprehensive clinical examination, ultrasound, colposcopic, hysteroscopic studies were conducted. Microbial flora analysis included bacterioscopic, bacteriological research methods with determination of sensitivity to antibiotics, and PCR method. The concentration of cytokines in the culture medium (supernatant) was determined by the enzyme immunoassay. Statistical data processing was performed using the Statsoft Statistica v6.0 and Microsoft Excel 97 software packages.
 Results. In the structure of gynecological diseases, 95.6% of patients with adenomyosis had inflammatory diseases of female genital organs, chronic herpes and cytomegalovirus infection was found in almost 93%. In women with adenomyosis, the II-III stage of vaginal cleanliness prevails with an “intermediate” type of biocenosis, which is characterized by a reduced content of lactobacilli, the presence of various types of morphotypes of gram-positive and gram-negative rods and cocci, and dysbiosis (49.4%), when a mixed bacterial flora prevails microflora, a high percentage of the combination of adenomyosis with chronic inflammatory diseases of the pelvic organs justifies the need to use antimicrobial therapy in this category of patients. When studying the cytokine profile of the cervical contents and endometrium, an increase in the concentration of the main proinflammatory cytokines IL-1β, IL-6, TNFα and a factor with chemotoxic properties IL-8 was found. The obtained data indicate that the development of adenomyosis is facilitated by a previous immuneinflammatory reaction, which is accompanied by a violation of the local cytokine balance.
 Conclusions. Microbiological examination of patients with adenomyosis revealed a violation of the microbial landscape of the reproductive tract in most of the examined. The obtained data indicate a high percentage of the combination of adenomyosis with chronic inflammatory diseases of the pelvic reproductive organs. Previous adenomyosis is an immunoinflammatory reaction, accompanied by a violation of the local cytokine balance. Increased cytokine activity and the presence of infectious agents are associated with the spread of adenomyosis.

The impact of chronic herpes virus infection on the bone system in children

Herpes virus infections currently are urgent problem in infectology, pediatrics and therapy. The feature of these diseases is the possibility of direct and indirect effects of the virus on various organs and systems: immune, nervous, endocrine, bone, in children in particular. The aim of the work is to summarize and systematize the available data on the impact of the chronic herpes virus infection on the bone system in children. Materials and methods: analysis of relevant Russian and international literature was carried out. Results: chronic herpes virus infections have pathological effects on bone system, in particular they can increase the risk of osteoporosis. Herpes viruses are involved in the pathogenesis of systemic inflammatory diseases, such as rheumatoid arthritis. Bone complications of herpes virus infections are the relevant issues. More research is needed to understand these complications more deeply.

Ambiguity effects of recombinant interferon α2B in an in vitro experiment on the expression of nuclear factor NF-KB, IFNα’βR AND IFNγR (CD119) receptors of neutrophilic granulocytes of patients with chronic herpes virus infections

Introduction. Neutrophil granulocytes (NGs) and the interferons system (IFN) play a leading role in antiviral immune defense. At the same time, patients suffering from atypical chronic active herpes-viral infections (ACA-HVI) often have neutropenia and impaired induced IFN/ and IFN production. It is possible that these patients have various disorders of intracellular signaling at all stages of IFN/ synthesis by immune system cells, including NG, which leads to IFN/ deficiency. An adequate level of nuclear factor NF-kB expression has positive effects on synthesis of IFN/ in the last stages of signaling. The disturbances in the expression of NF-kB can lead to defects in the synthesis of IFN/.
 The aim: to clarify the features of nuclear factor NF-kB, membrane receptors IFN|R and IFNR (CD119) expression on NG of patients with AСA-HVI, with the subsequent assessment of the recombinant IFN2b (recIFN2b) effects on them in the experimental system in vitro.
 Materials and methods: 25 patients with ACA-HVI of both sexes aged 23-64 years were included in the main group of the study (MSG). In the complex of the study for the detection of herpes-viral infections: the method of serodiagnosis (ELISA), for the detection of the genome of viruses - PCR-RT. 407 blood samples were examined in the in vitro system. Flow cytofluorimetry was used to estimate the amount (%) of NG expressing NF-kB, IFN|R, IFNR, and their expression levels before and after incubation with recIFN2b. Adequate statistical methods were applied.
 Results: In patients suffering from ACA-HVI, a deficiency of induced IFN and IFN production was detected, due to a decrease in the expression density of nuclear factor NF-kB NG, as well as a violation of the expression of membrane receptors IFN|R, IFNR. RecIFN2b in the in vitro system had a mixed effect on the expression of NF-kB, IFN|R and IFNR NG.
 Conclusion: The deficiency of induced IFN and IFN production in patients with ACA-HVI is associated with variable changes in NF-kB, IFN|R and IFNR NG expression. RecIFN2b in the in vitro system has an ambiguous effect on altered expression of NF-kB, IFN/ and IFN NG, which appears to depend on the innate or adaptive nature of these disorders.

Features of Interferon Status and Methods of Its Correction in the Treatment of Patients with Atypical Chronic Active Herpesvirus Infections.

It is known that the imbalance of the IFN system is the cornerstone of the immunopathogenesis of atypical chronic active herpesvirus infections and is often associated not only with congenital, genetically determined defects, but also with acquired system dysregulation at different levels: receptor, molecular, at the level of a nuclear transducer of signal transmission. Based on the studied features of immunopathogenesis and the revealed disturbances in the antiviral immune defense system and the IFN system in patients with atypical chronic herpes viral infections, an integration program for correcting IFN status was developed, its clinical and immunological effectiveness was evaluated. Improved effectiveness of complex rehabilitation of immunocompromised patients with atypical chronic active herpes viral infections was demonstrated.

Positive effects of recombinant interferon α2b on the phenotype of CD16<sup>+</sup>INFα/βR1<sup>-</sup>CD119<sup>+</sup>, CD16<sup>+</sup>INFα/βR1<sup>+</sup>CD119<sup>-</sup> neutrophil granulocyte subset in patients with post-COVID syndrome and herpesvirus infections

Post-COVID syndrome (PCS) is a multisystem inflammatory condition with manifestations of chronic fatigue syndrome (CFS) and cognitive disorders (CD), along with reactivation of chronic herpesvirus infections (HVI). The PCS manifestations require studying the molecular mechanisms associated with the production of IFN and receptor functions of neutrophil granulocytes (NG), which is relevant and promotes the search for immunotherapeutic strategies in patients with PCS. Our objective was to study the in vitro effects of recombinant interferon α2b (recIFNα2b) on the phenotype of CD16+IFNα/βR1-CD119+, CD16+IFNα/ βR1+CD119+ subsets and functional activity of NG in patients with post-COVID syndrome and herpesvirus infections. Materials and methods: 45 patients (24-60 years old) with PCS and HVI (HSV 1, EBV, HHV6, CMV) comprised the study group 1 (SG1). A questionnaire was conducted to assess the severity of PCS symptoms using a point scale. We performed a study of the content and phenotype of NG subsets, i.e., the CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119-, CD16+IFNα/βR1+CD119+ subpopulation, phagocytic and NADPH oxidase function of NG before and after in vitro incubation with recIFNα2b (50 IU/ µL, for 60 min, at 37 °C) in the study group 1a (SG1a). The comparison group (CG) of 30 volunteers examined during the pre-COVID period. Results: We revealed more pronounced clinical manifestations of CFS and CD in SG1 patients with mixed HVI, than in mono-HVI cases. Increased expression density of all receptors was registered on CD16+IFNα/βR1+CD119-NG and CD16+IFNα/βR1-CD119+ NG, thus suggesting the NG activation with initiation of cytotoxicity or NETosis, a decrease in phagocytic function and intensity of NADPH oxidase activity with depletion of NG reserve capacity in SG1. We have obtained some data on the positive effect of recIFNα2b in vitro (SG1a), e.g., decreased CD16 expression density and enhancement of IFNα/βR1 receptor expression in the CD16+IFNα/βR1+CD119- subset. In the CD16+IFNα/βR1-CD119+ subset, we have found persistence of increased MFI CD16 and MFI CD119 receptors, restoration of defective NG phagocytic function and reduced excessive activity of NADPH oxidases. Conclusion: The positive effects of the recIFNα2b influence on deficient function of NG in PCS patients suggest an oppoptunity of using immunotherapy with a recIFNα2b-based drug, combined with highly active antioxidants for treatment of various PCS manifestations including CFS, CD, HVI, thus, probably, ensuring adequate functioning of antiviral and regulatory mechanisms of the immune system.

Hematopoietic stem cells and betaherpesvirus latency.

The human betaherpesviruses including human cytomegalovirus (HCMV), human herpesvirus (HHV)-6a and HHV-6b, and HHV-7 infect and establish latency in CD34+ hematopoietic stem and progenitor cells (HPCs). The diverse repertoire of HPCs in humans and the complex interactions between these viruses and host HPCs regulate the viral lifecycle, including latency. Precise manipulation of host and viral factors contribute to preferential maintenance of the viral genome, increased host cell survival, and specific manipulation of the cellular environment including suppression of neighboring cells and immune control. The dynamic control of these processes by the virus regulate inter- and intra-host signals critical to the establishment of chronic infection. Regulation occurs through direct viral protein interactions and cellular signaling, miRNA regulation, and viral mimics of cellular receptors and ligands, all leading to control of cell proliferation, survival, and differentiation. Hematopoietic stem cells have unique biological properties and the tandem control of virus and host make this a unique environment for chronic herpesvirus infection in the bone marrow. This review highlights the elegant complexities of the betaherpesvirus latency and HPC virus-host interactions.

Peculiarities of post-viral chronic fatigue syndrome associated with mild cognitive decline in patients with atypical chronic active herpesvirus infections

According to modern ideas, changes in the functioning of the immune system affect the immune processes in the nervous system, contributing to the development of neuro-immuno-inflammation and thereby indirectly affect the rate of progression of neurodegenerative processes. The aim of our study was to investigate the prevalence of post-viral chronic fatigue syndrome and cognitive impairment (aMCI) among patients with atypical, chronic active herpesvirus infections (ACA-HVI).Under our supervision were 126 patients of both sexes aged 18 to 60 years with ACA-HVI.It was established that mono-EBV infection affects 27.7%; mixed EBV infection is observed in 72.3% of patients. When assessing cognitive functioning using CGI, MMSE scales, the incidence of aMCI was found to be 68.3%: with mixed HVI — 87.4%, with mono HVI — 38.8%. During the study, significant limitations were identified in the use of standard scales due to the impossibility of conducting a comprehensive assessment of clinical status parameters and cognitive dysfunctions, as well as correlation of these parameters and assessment of dynamics of the immunocorrection. To achieve this goal the Scale of assessment of the criterion clinical symptoms of patients with ACA-HVI with CFS was used. It was shown that in mixed-HVI, the severity of symptoms exceeded the severity of symptoms of patients with mono-HVI and was 52.7 (43.1-62.2) and 38.0 (31.9-42.8) points, respectively (p > 0.05). Thus, it was found that patients suffering from mixed HVI have more pronounced, severe manifestations of CFS and aMCI, which are 1.5 times higher than similar manifestations in patients with mono-HVI, significantly reducing the quality of life of these patients, worsening their social adaptation.Prolonged persistence of herpes viruses in immune-compromised people creates conditions for constant antigenic stimulation and immune imbalance with the onset of secondary immunodeficiency or clinical manifestation of existing primary disorders in the immune system, which creates the prerequisites for the development of neuro-immuno-inflammatory changes in nervous system, followed by the formation of clinical manifestations of ME/CFS with different cognitive impairments that may be classified as aMCI.

Diagnostically significant changes in subsets CD11b<sup>+</sup>CD64<sup>-</sup>CD32<sup>+</sup>CD16<sup>+</sup>, CD11b<sup>+</sup>CD64<sup>+</sup>CD32<sup>+</sup>CD16<sup>+</sup> neutrophilic granulocytes of immunocompromised women with chronic infectious and inflammatory diseases of the genital tract of various etiologies

Chronic pelvic inflammatory disease (PID) in women remains a problem due to the importance of medical consequences. The study of the receptor apparatus of neutrophilic granulocytes (NG) involved in anti-infective protection in diseases of various etiologies seems to be relevant. Aim: to clarify the features of variants of quantitative and phenotypic changes in subsets of NG CD11b+CD64-CD32+CD16+, CD11b+CD64+CD32+CD16+ of immunocompromised women during exacerbation of chronic PID of various etiologies. We were tested women 20-40 years: Study Group 1 (SG1, n = 20) – chronic PID during the exacerbation with mono- or mixed latent/recurrent various viral infections (chronic herpes-virus infections, papillomavirus infection, recurrent ARVI); Study Group 2 (SG2, n = 30) – chronic PID of bacterial etiologies; Comparison Group (CG)– 20 healthy women. The number of subsets CD11b+CD64-CD32+CD16+NG (major) and CD11b+CD64+CD32+CD16+NG (minor), receptor expression density (MFI) was determined (FC500, USA). It was found that in PID during the period of exacerbation, diagnostically significant differences in the subset composition of NG were revealed. We got a decrease in the CD11b+СD64-СD32+СD16+NG subset and in 7,6 times increase in the CD11b+CD64+CD32+CD16+NG subset in SG2 with chronic PID of bacterial etiology, in contrast to chronic PID occurring in combination with recurrent/persistent viral infection SG1. Negative transformation of NG subsets is associated with a predominant decrease in the level of expression of the activation CD16. The absence of an adequate response to the infectious and inflammatory process was revealed – the absence of an increase in the expression of the activation CD11b in the major subset in SG1, as well as in the minor subset in groups SG1 and SG2. In the major subset of NG in groups SG2 a decrease in the expression of the activation marker CD11b. In the various viral infections and PID (SG1), in the negatively altered minor subset of NG we got a decrease of expression of CD16, an increase of expression of CD64 and CD32. Determination of subsets of CD11b+СD64-СD32+СD16+, CD11b+CD64+CD32+CD16+NG and their phenotype can be used as diagnostic markers for the differential diagnosis of PID of viral and bacterial etiology, and for the development of new methods of targeted immunomodulatory therapy.

Clinical features and treatment of activated PI3K-delta syndrome in Peruvian children: A case series

IntroductionActivated phosphoinositide 3-kinase δ (PI3Kδ) syndrome (APDS) is an inborn error of immunity (IEI) caused by a gain-of-function mutation in the PIK3CD gene. It is characterized by recurrent respiratory infections, chronic herpes infections, lymphoproliferation, mucosal lymphoid hyperplasia, enteropathy and immune dysregulation. This condition can be misdiagnosed with other diseases, such as lymphoma or other IEI. Additionally, limited knowledge of APDS in physicians could generate delay in diagnosis, treatment and lead to worse outcomes, especially in developing countries. This study aimed to describe the clinical features and treatment of APDS in Peruvian children evaluated in two centers. MethodsWe retrospectively analyzed data from eight patients with APDS evaluated in the two main Peruvian clinical immunology centers. Demographics, clinical features, laboratory analysis, and treatment data were included. ResultsAll patients were female. At the onset of symptoms and diagnosis, the mean age was 1.4 (SD: 0.98) and 6 (SD: 2.1) years, respectively. The mean delay in diagnosis was 4.5 (SD: 2.7) years. No consanguinity was reported, and only one patient had a family history of IEI. The mortality rate was 25% (2 patients). The main clinical manifestations were lymphadenopathy and hepatomegaly (100%), chronic diarrhea and recurrent pneumonia (87.5%), and splenomegaly (62.5%). All the patients had E1021K mutation. Seven patients (87.5%) had elevated IgG and 50% had elevated IgM. Regarding treatment, all the patients received intravenous immunoglobulin (IVIG), 50% were treated with everolimus and only one patient received hematopoietic stem cell transplantation. ConclusionLymphadenopathy, hepatosplenomegaly, chronic diarrhea, pneumonia, and IVIG treatment were the main features. Our cohort had limited treatment options with calcineurin inhibitors and transplantation and laboratory testing such as lymphocyte population and functional antibody tests. Although genetic testing was done on all the patients, it was supported by international collaboration. Therefore, further studies are needed to evaluate the diagnosis and management impact on patients with APDS in our setting.

Two cases of bloodstream infections associated with opportunistic bacterial species (Enterococcus hirae and Enterobacter xiangfangensis) in companion animals

BackgroundBloodstream infections are a matter of concern in small animal veterinary practice. Few reports are avaiable, especially regarding the role of opportunistic bacteria in becoming infectious. This report aims to add to the current veterinary literature on two opportunistic bacterial species (Enterococcus hirae and Enterobacter xiangfangensis) associated with bloodstream infections in small animals admitted to the Bologna University Veterinary Hospital.Case presentationIn the first case, a 15-year-old, immunocompromised, cardiopathic dog was admitted to the hospital for anorexia and diarrhea. The patient had a history of previous surgery and hospitalization. After three days, hyperthermia, leukopenia and hyperlactatemia were recorded, and blood culture revealed positivity for Enterococcus hirae, identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The patient’s general conditions progressively worsened, and the patient was euthanized. In the second case, a 2-year-old cat with chronic ocular herpesvirus infection and hypertrophic cardiomyopathy was admitted to the hospital for anorexia and hyperthermia. The cat was hospitalized one week before and received antimicrobial treatment for urinary tract infection by Staphylococcus felis. Hypokalemia and lymphopenia were also diagnosed. The patient progressively improved and was discharged after three days. On the same day, blood culture taken at admission revealed positivity for Enterobacter xiangfangensis, identified using MALDI-TOF MS. After five days, the patient returned with neurological symptoms, hypothermia and bradycardia, and was euthanized.ConclusionsIn small animal veterinary practice, the impact of opportunistic bacterial agents (such as E.hirae and E.xiangfangensis) on bloodstream infections remains unclear. As in human medicine, they can be contracted in every healthcare setting and considered hospital-acquired infections. In this report, we highlighted the threat they pose especially in patients with multiple risk factors. Rapid and accurate diagnostic tools (such as MALDI-TOF MS) could be particularly important for reducing the severity of the infections.

Atypical chronic active herpesvirus infections: Etiological structure, frequency of occurrence, clinical syndromes associated with them

BACKGROUND: Every year, a steady, progressive increase in the number of atypical chronic, active forms of infections caused by herpesviruses is recorded. Diagnosing and selecting adequate therapeutic strategies for treating these infections present significant difficulties for physicians due to the polysyndromicity and many clinical manifestations.
 AIMS: This study determines the prevalence of atypical chronic, active infections caused by herpesviruses among patients infected with herpesvirus infections and studies the etiological structural features and clinical manifestations/criteria and signs of atypically occurring chronic forms of herpes viral infections.
 MATERIALS AND METHODS: Under our supervision at the Clinical and Diagnostic Center Medsi in Belorusskaya (Moscow), 98 patients of both sexes aged 23 to 60 years suffering from atypical chronic, active forms of infections caused by herpesviruses comprised the herpesviruses group. The comparison group consisted of 30 conditionally healthy subjects comparable in sex and age to patients. In addition to traditional methods (history collection, physical examination, general blood test, and others), serodiagnostic methods with ELISA were used to detect herpesvirus infections. ELISA was also used for detecting the genome of viruses in biomaterials. The study was approved by the ethics committee, and all patients received informed consent to participate in the study. Statistical analysis was performed using adequate methods.
 RESULTS: The study of the etiological structure of herpesvirus infections in patients with atypical chronic, active forms of infections caused by herpesviruses, mixed herpesvirus infections were shown to occur in 83.4% of patients, and mono herpesvirus infections in 16.6% of cases. It was shown that the EpsteinBarr virus was the dominant virus among patients with mono and mixed herpesvirus infections. A high rate of EpsteinBarr virus DNA detection was demonstrated in saliva (84.2%), posterior pharyngeal wall scrapings (73.5%), tonsils (42.9%), urine (12.6%), and blood (8.3%) is a marker of the high replicative activity of the virus. The primary clinical syndromes associated with mono and mixed atypical chronic and active forms of infections caused by herpesviruses were identified.
 CONCLUSIONS: This study identified and quantitatively assessed the viral load associated with the severity of the course and clinical manifestations of atypical chronic, active forms of infections caused by herpesviruses. Clarifying the features of clinical manifestations and syndromes in patients suffering from various mono and mixed herpesvirus infections will allow us to outline the goals for the further development of an adequate diagnostic algorithm for these atypical forms of herpesvirus infections and the concept of targeted, personalized etio- and immunopathogenetic therapy.

Chronic recurrent urticaria in a patient with recurrent herpes labialis: complete remission after administration of aciclovir and antihistamines.

Chronic recurrent urticaria, occurring in the context of infections, represents a major challenge for clinicians. Chronic genital herpes infection has been perceived in the literature as a possible trigger of chronic recurrent urticaria. The administration of a systemic corticosteroid regimen in these cases has no long-lasting effect and the subsequent relapses are difficult to control. In these cases, treatment of the urticaria with antihistamine (as monomedication) is often not sufficient and does not suppress the symptomatology. The administration of acyclovir or valacyclovir according to a specific therapeutic regimen as monotherapy or in combination with antihistamine has been shown to be quite effective. The doses of this administration vary and can be tailored to clinical symptomatology. We present a 41-year-old female patient with chronic recurrent urticaria associated with angioedema and bronchospasm, in whom her herpes genitalis was found to be concurrent within the history and clinical examination. According to the history, the herpes was also recurrent and dated back about a year. Chlamydia trachomatis infection was also found, with serological findings corresponding to vaginal discomfort. Treatment with acyclovir 400mg thrice daily for an initial period of 7 days in combination with desloratadine 5mg daily was started as we observed complete remission of the urticarial rash. Due to worsening vaginal discharge, it was decided to temporarily discontinue systemic acyclovir therapy and treatment for the chlamydial infection was initiated with doxycycline 100 mg twice daily for 21 days. Already on day 1 after stopping acyclovir, a severe relapse with generalization of the urticarial rash was observed. Control of symptomatology was achieved by reintroduction of acyclovir according to the regimen in combination with antihistamine. Chronic infections are one of the common causes of chronic recurrent urticaria with a tendency to generalization and possible complications such as angioedema and bronchospasm. An academic, analytical approach to patients and the consistent exclusion of each possible trigger for chronic recurrent urticaria often guarantee the success of subsequent treatment.

Алгоритм клинико-иммунологической диагностики вариантов патологических иммунофенотипов, ассоциированных с атипичными хроническими активными герпесвирусными инфекциями

An imbalance of different parts of the antiviral immune response system associated with deficits of type I and type II interferons (IFN) is commonly observed in atypical chronic active herpesvirus infections (ACA-HVI). Objective. To develop a clinical and immunological diagnostic algorithm for detecting the pathological immunophenotypes of ACA-HVI. Patients and methods. A total of 263 male and female patients aged 23–70 years with ACA-HVI were examined. The study included the following methods for herpesvirus detection: serodiagnostics and a real-time polymerase chain reaction (RT-PCR). The cytokine profile (interleukins IL-1β, IL1Ra, IL-6, IL-17, TNFα) and the IFN system were analyzed by an enzyme-linked immunosorbent assay (ELISA). The study was approved by the Ethics Committee, and an informed consent was obtained from all patients. Results. To assess the clinical signs of immunosuppression in patients with ACA-HVI, we formulated de novo clinical criteria of class I and class II and used them in the study. Based on the three main pathological immunophenotypes identified, we created an algorithm for the clinical and laboratory diagnosis of ACA-HVI. It was shown that determining the serum levels of cytokines IL-1β, IL-6, IL1Ra and tumor necrosis factor-α is the most optimal option to assess the level of systemic inflammation and neuroimmune inflammation. Conclusion. The findings of the immunological study allowed us to identify variants of pathological immunophenotypes, which are characterized by the variability of the detected disorders. The revealed disorders in the immune system and IFN system in each pathological immunophenotype are associated with certain ACA-HVI manifestations, typical of study groups 1, 2, 3. Key words: herpesvirus infections, neutrophil granulocytes, interferon system, pathological immunophenotypes

Интерферонопатии при атипичных хронических активных герпесвирусных инфекциях: тактика проведения коррекции дефицита интерферонов

A cytokine imbalance and interferonopathies associated with deficits of type I and type II interferons (IFN) are often observed in atypical chronic active herpesvirus infections (ACA-HVI). Objective. To study the features of IFN system functioning and cytokine profile in patients with ACA-HVI before and after the immune system correction integrative program (ISCIP) including local and systemic interferon therapy. Patients and methods. A total of 39 male and female patients aged 23–70 years with ACA-HVI characterized by isolated disorders in the IFN system were examined. The study included the following methods for herpesvirus detection: serodiagnostics and a real-time polymerase chain reaction (RT-PCR). The cytokine profile (interleukins IL-1β, IL1Ra, IL-6, IL-17) and the IFN system were analyzed by an enzyme-linked immunosorbent assay (ELISA). The study was approved by the Ethics Committee, and an informed consent was obtained from all patients. Results. Patients with ACA-HVI had antiviral immune response disorders. Against the background of serum IFN deficiency, induced and spontaneous production of IFNα and IFNγ, a functional impairment of cytokine profile with predominant hyperproduction of proinflammatory cytokines IL-1β, IL-6, tumor necrosis factor-α were revealed. Positive clinical and immunological effects of ISCIP were demonstrated. It was shown that IL1Ra can be used as a diagnostic biomarker to monitor the course and outcome of ACA-HVI. Conclusions. The developed ISCIP with the inclusion of targeted local and systemic interferon therapy for patients with ACAHVI demonstrated high clinical and immunological efficacy against the background of the confirmed absence of anti-IFNα autoantibodies. Key words: herpesvirus infections, interferonopathies, neutrophil granulocytes, interferon system

Morphological structure of cotyledons with non-contrasted blood vessels in the placenta of women who had an exacerbation of mono- and mixed-cytomegalovirus infection in the second trimester of pregnancy

Aim. To assess the morphological structure of cotyledons with non-contrasted blood vessels in the pla­centas in women who had an exacerbation of mono- and mixed cytomegalovirus infection (CMVI) in the second trimester of pregnancy.Materials and methods. The histometric parameters of the villi were studied with different visualization on X-ray phlebograms of blood vessels in 112 cotyledons of placentas in women with pregnancy, uncomplicated and com­plicated by mono- and mixed-CMVI (combination of CMVI and reactivation of chronic herpes virus infection type 1 HSV). The first group (control) included 36 cotyledons with a clear contrasting of the vascular bed in the placentas from women with an uncomplicated gestational period. The main group was represented by 76 cotyledons: subgroup 2a - 24 cotyledons, in which the bloodstream was not detected, in placentas from patients with exacerbation of mono-CMVI as­sociated with chronic subcompensated placental insufficiency (CSPI); subgroup 2b - 16 cotyledons with blood vessels not diagnosed by X-ray phlebography, in placentas from women with mixed CMVI infection, initiating the development of CSPI; subgroup 3a - 21 cotyledons with non-contrasted bloodstream in placentas from patients with exacerbation of mono-CMVI and chronic decompensated placental insufficiency (CDPI); subgroup 3b - 15 cotyledons with a similar an­giographic picture in placentas from women who had an exacerbation of mixed CMVI in the second trimester of gestation and CDPI. Identification of cotyledons with a clear visualization of blood vessels and with a non-contrasted vascular bed in the marginal part of the placenta was carried out by dosed administration of minium on linseed oil (1:3) through the vein of umbilical cord and assessment of the angiographic picture on X-ray, which were obtained using the RUM-20M apparatus with X-ray image amplifier Sapphire (Russia). For histological analysis, tissue pieces were taken in the same places before and after injection of the contrast mass into the bloodstream of the organ, on the damaged areas of which clamps were applied.Results. In the first group, the cotyledons had clear contours of the blood vessels and the vein draining them. Avascular villi (AV) accounted for 2.05±0.22%, villi with syncytiocapillary membranes (SCM) - 33.5±2.41%, villi with one (1) SCM - 77.1±2.07%, with two (2) SCM - 20.9±1.98% and with three (3) SCM - 1.42±0.09%. In subgroup 2a, compared with the first group, there was a deformation of the lumen of the vein of the stem villi of the I and II order, in which erythrocyte aggregates, single leukocytes and rarely thrombi were determined, as well as signs of edema and partial desquamation of endotheliocytes, fibrinoid and inflammatory changes in the vessel wall. The number of AV increased (by 6.87 times, p<0.001) and the number of villi with SCM decreased (by 2.05 times, p<0.001) with no significant differences between villi with 1, 2, and 3 SCM. In subgroup 2b, in comparison with subgroup 2a, mod­erate folding of the endothelium, areas of its partial desquamation, AV (1.39 times, p<0.01) and villi with 2 SCMs (1.44 times, p<0.05) were more common against the background of a decrease in the concentration of villi with 1 SCM (by 1.19 times, p<0.05). In subgroup 3b, a greater number of vessels were found that did not provide outflow of blood from cotyle­dons (12, p<0.001) and had an arcuate shape (10, p<0.001) compared with subgroup 3a, where their number was, respec­tively, 3 (p<0.001) and 2 (p<0.001). A more pronounced folding of the endothelial lining was determined, as well as ag­gregates, neutrophils, lymphocytes and thrombi in the lumen of the vessels. There was an increase in the percentage of villi with 1 SCM (by 1.13 times, p<0.05), as well as a decrease in the frequency of occurrence of villi with 2 SCM (by 1.67 times, p<0.05) and 3 SCM (by 2.13 times, p<0.001). In cotyledons of subgroup 3b, in contrast to those of subgroup 2b, blindly ending vessels were more common (2 times, p<0.05), arcuate course of veins (2.5 times, p<0.05), lumen de­formity, hyperchromia of the nucleus and total desquamation of cells, thrombi with calcification, complete and partial obliteration of the arterial lumen, inflammation, fibrinoid degradation of the muscular layer of vessels and the deposition of calcium salts in it, as well as AV (1.29 times, p<0.05) and villi with 1 SCM (1.40 times, p<0.001); less often villi were detected with SCM (1.35 times, p<0.05), with 2 SCM (2.78 times, p<0.001) and 3 SCM (1.57 times, p<0.05).Conclusion. In women with exacerbation of mixed CMVI in the second trimester of gestation and CDPI, in contrast to patients with reactivation of mono-CMVI leading to CDPI, angiodestructive and angioobstructive processes, as well as inhibition of angiogenesis, are more pronounced in cotyledons with non-visualized blood vessels. This increases vascular resistance and blocks the flow of the contrast mass during its dosed injection into the vascular bed of the placenta.

Etiological factors of voiding disorders in females

Background. Voiding disorders in females are a serious medical and social problem. Various urological, gynecological, and neurological factors contribute to the development of these disorders.
 Aim. To study etiological factors of voiding disorders in females.
 Materials and methods. A total of 52 women 19 to 81 years of age with various voiding disorders were inspected. Examination algorithm included collection of a comprehensive medical history, filling in voiding diaries, gynecological exam, various laboratory tests (including PCR-diagnostics of urethral scrapings; ELISA tests for IgM and IgG to herpes simplex virus types 1 and 2, Varicella Zoster virus, EpsteinBarr virus, cytomegalovirus; vaginal discharge culture for flora and sensitivity to antibiotics, with mandatory determination of the lactobacilli count) and diagnostic methods, as well as consultations by related specialists.
 Results. Chronic herpesvirus infection contributed to urinary dysfunction in 28 (53.8%) women, bacterial urinary tract infection in 15 (28.8%), bacterial vaginosis in 15 (28.8%), papillomavirus infection in 12 (23.1%), and interstitial cystitis in 10 (19.2%) patients. Thirty-five (76.9%) women had several simultaneous factors that contributed to the development of voiding disorders.
 Conclusion. The combination of disorders of the vaginal microflora with viral and/or bacterial infections most often contributes to the development of voiding disorders.