Chronic Pediatric Immune Thrombocytopenia Is Not Associated With Herpes Virus Infection Status

Background: Immune thrombocytopenia (ITP) is characterized by non-chronic (transient, <12 months) and chronic (≥12 months) decline in the number of platelets. Herpes virus infections have been shown, in many studies, to be associated with the development of ITP. However, it remains unclear whether the herpes virus infection status is associated with the chronic ITP.Methods: We reviewed 480 primary pediatric patients with ITP in the period from January 2017 to December 2019. The prevalence of herpes virus antibodies including the Cytomegalovirus (CMV), Herpes simplex virus 1 (HSV-1), Herpes simplex virus 2 (HSV-2), and Epstein Barr virus were recorded. The levels of serum complement C3 and C4, T (CD3+, CD4+, CD8+), B (CD19+) lymphocytes, and natural killer (CD16+ 56+) cells were also analyzed. Multivariate analysis was used to evaluate the associations between chronic ITP and herpes virus infection status.Results: Compared with non-chronic, patients with chronic ITP had older age (≥3 years), lower levels of hemoglobin and complement C3, and lower probability of CMV and HSV-2 infections (IgM positive; p < 0.05). Patients with herpes virus infection had lower serum platelet counts (p < 0.001), lower complement C3 levels and lower CD4+/CD8+ cells ratio (p < 0.05). Furthermore, platelet counts were positively correlated with CD4+/CD8+ cells ratios (r = 0.519; p = 0.0078), and negatively correlated with T cells (CD3+: r = −0.458, p = 0.0213; CD8+: r = −0.489, p = 0.0131). Multivariate analysis showed that age (OR, 1.644; 95%CI, 1.007–2.684; p = 0.047) was an adverse risk factor for chronic ITP and CMV IgM positive (OR, 0.241; 95%CI, 0.072–0.814; p = 0.022) had lower risk of chronic ITP development, while other herpes virus infection statuses and clinical features were not.Conclusion: Although herpes virus infections were associated with the onset of ITP, our findings indicated that herpes virus infection status might not be a risk factor for chronic ITP.

PECULIARITIES OF EFFECTOR LYMPHOCYTES SUBPOPULATIONS IN PATIENTS WITH UVEAL MELANOMA DURING ACTIVATION AND CHRONIC PROGRESS OF HERPESVIRAL INFECTION

The immune response to any antigen includes the induction of effector and regulatory T lymphocytes. In tumors, an imbalance in the subpopulation of lymphocytes is noted. Ophthalmotropic pathogens of herpes virus infections play a role in the etiopathogenesis of some oncological processes in the tissues of the eye. Their “cancer modulating” role is to regulate the functions of immunocompetent cells by viruses and reprogram it in the direction of greater progression of tumor growth. The aim of our study was a comparative analysis of the content of effector subpopulations of blood lymphocytes in patients with uveal melanoma during activation and the chronic course of herpes virus infection. The study involved 141 people: with uveal melanoma 70 patients, with corneal ulcers and involvement of the uveal tract - 38 patients and 33 healthy donors. Immunophenotyping was performed by flow laser cytofluorimetry using a monoclonal antibody system to differentiate lymphocyte subpopulations. IgM and IgG antibodies to herpes virus infections were determined in an enzyme-linked immunosorbent assay (ELISA) on an automatic ELISA analyzer "Lazurite" (USA) with diagnostic kits of CJSC Vector-Best (Koltsovo). The results of the study showed that the absolute number of blood lymphocytes (CD45 +) in patients with uveal melanoma, regardless of the presence of active or chronic herpes virus infection, did not differ from the values in healthy donors. In patients with corneal ulcers involving the uveal tract, an increase was noted. A decrease in the relative and absolute content of T cells (CD3+) was revealed in patients with uveal melanoma during activation and the chronic course of herpes virus infection. An increase in the absolute content of CD3 + was noted in patients with active and chronic herpesvirus infection in patients with inflammatory lesions of the eye. There was no difference in the relative and absolute contents of the subpopulation of CD3 + CD4 + helpers / inducers upon activation of herpes virus infections in tumor. With corneal ulcers - a significant increase in the absolute content of CD3 + CD4 + helpers / inducers with active and chronic infection. In chronic herpes virus infection, an increase in the relative and absolute number of cytotoxic T-lymphocytes (CD3 + CD8 +) with uveal melanoma was revealed. With active infection, a decrease in the relative number of T-lymphocytes (CD3 + CD8 +) in the tumor was noted. With corneal ulcers involving the uveal tract, an increase in the absolute content during activation and chronic infection and a decrease in the relative content compared to the norm were determined. When analyzing the “double positive” T cells in a tumor, an increase in relative and absolute indices was observed in chronic and active infection with herpes viruses. The same trends were observed with inflammatory eye diseases. Analysis of the content of B-lymphocytes (CD19 +) in blood with melanoma showed an increase in the absolute number during activation and the chronic course of herpes virus infection. When analyzing the indicators of the absolute and relative content of B-lymphocytes (CD19 +) with inflammation of the membranes of the eyes, their increase was revealed regardless of activation or chronic infection. The relative number of natural killers (CD16 + CD56 +) of blood with uveal melanoma increased with infection activation. In inflammatory eye diseases, a decrease in the relative content of natural killers (CD16 + CD56 +) was found in chronic infection and active herpes virus infection. With melanoma, no changes in the index of the ratio CD4 + / CD8 + were detected, with ulcerative lesions of the cornea involving the uveal tract, its increase was noted in acute and chronic infections with herpes virus infections. (p <0.05). Thus, our own studies suggest a deep suppression of the immune system of the body of patients with uveal melanoma, which does not allow the activation of antiviral protection. Tumor causes multidirectional shifts in the relative and absolute content of lymphocytes during activation and chronic infection with herpes virus infections. The results obtained are important for the development of personalized approaches to the prognosis and treatment of patients with uveal melanoma.

Identification of A New Biomarker For Herpes Zoster Infection in Rheumatoid Arthritis

Herpes zoster (HZ) is known as a side effect of using biologics in rheumatoid arthritis (RA). Incidence of this side effect may be different depending on genetic factors because susceptibility to HZ infection varies by race. Here, we analyzed the statistical relationships of whole genome single nucleotide polymorphisms (SNPs) with HZ infection in biologics-treated RA patients.The subjects were 321 Japanese female patients (including 56 herpes virus infected patients) of RA using biologics. The relationships of 302,814 SNPs with HZ infection were analyzed using case-control analyses by Fisher’s exact tests. We picked up SNPs (P < 10-8) significantly associated with HZ infection. Then, herpes infection was compared among the genotypes using a multivariate logistic regression analysis adjusted for onset age of RA.Rs10774580 located in 2’-5’-oligoadenylate synthetase like gene (OASL) was significantly associated with herpes virus infection. The minor allele homozygous carrier was positively associated with herpes virus infection in multivariate analysis.We for the first time showed a significant relationship between a genetic factor and HZ infection among RA patients. Rs10774580 may be one of the biomarkers for HZ infection.

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.

INCIDENCE OF CONFIRMED MANIFEST VIRAL INFECTION IN CASES OF ISCHEMIC STROKE, DEPENDING ON THE SEASON

Relevance. Seasonal differences in stroke incidence are associated with various physical factors, some associated with pathophysiological changes in the body leading to ischemic stroke. At the same time, there is little research into the differences in the frequency of detection of the genome of herpes virus infection and influenza virus, depending on the season. Objective. Investigate the existence of associations between the season and the frequency of virologically confirmed herpezoid infection manifested by herpezoid virus and influenza virus of patients with ischemic stroke. Materials and methods. In the period from 01.01.2017 to 31.12.2017. during the year at the bases of the neurological and department of the Alexandrovsky Clinical Hospital Centre. Kyiv conducted a study of 144 cases of cerebral ischemic stroke with the medium-severe neurological deficit: 78 (54.2%) women and 66 (45.8%) men, with an average age of 63.1 0.8 years. Neurological examination and identification of the genome of herpes viruses and influenza virus, performed with the help of a polymerase chain reaction (PCR), was carried out monthly in 12 patients hospitalized by ambulance. The account was taken of the presence of a viral manifestation if it preceded the stroke within 2 weeks. Results. 36 cases were examined each season. The ratio of men to women was 45.8 / 54.2 in total and did not differ between seasons, p = 0.514. 32 (22.2 percent) patients have demonstrated a viral infection. In winter, 11 (34.4 percent), in spring, 7 (21.9 percent), in summer, 4 (12.5 percent), in autumn, 10 (31.3 percent), and all p. At the same time, during the summer period, the incidence of viral manifestation was definitely lower than in winter, p = 0.042. Virus genomes were found in 12 (33.3 percent) patients in winter, 7 (19.4 percent) in spring, 5 (13.9 percent) in summer, 12 (33.3 percent) in autumn and p = 0.131 rice. During the summer period, the frequency of PCR-confirmed herpes virus infection was definitely lower than in the winter-autumn season, p = 0.033. Conclusions. Viral manifestations of herpes and influenza virus in the blood are more frequent during the winter and autumn periods; the frequency of detection indicated viral infection from October to January is reliably higher than the rate from March to August inclusive. In 25.0 percent of patients with ischemic stroke, the genome of herpes viruses and influenza virus in the blood is detected. In the event of a demonstration, the frequency of the above-mentioned viral infection is clearly higher (90.6 percent as against 9.4 percent) than that of patients without it.

The effect of Esberitox on the functional activity of 2D-positive natural killer cells (NKG2D) and interferon status in patients with recurrence of herpes simplex virus infection

The aim of the study was to assess changes in the functional activity of 2D-positive natural killer cells (NKG2D) and interferon status in patients with recurrence of herpes virus infection under the influence of Esberitox.Materials and methods. We studied 30 patients aged 22-45 years old with frequent relapsing labial herpes, previously verified by PCR. The frequency ofexacerbations at the time of the study was 6 (5.3 ± 1.4) and more than once a year. Patients were initially examined within 24 hours of the appearance of rashes.The expression of CD314 on the surface of CD3-CD56 lymphocytes was determined by flow cytometry(EPICS XL, Beckman Coulter). The content of cytokines (α-and γ-IFN) in supernatants was determined by enzyme immunoassay (IBL, Germany). Patients took Esberitox tablets, 2 tablets x 3 times a day, and daily1-gram tablets of Valacyclovir for 7 days. The control group consisted of 10 patients receiving only tablet valacyclovir.Results. As a result of the study, it was found that the final leveling of symptoms with combined treatment with Esberitox was noted by the end of 5-6 days, with basic treatment - by 9-10 days. There was a decrease in the relapse rate (3 patients) during the observation period within 2 months after the end of the course of treatment.In patients receiving basic Valacyclovir therapy, relapses during the observation period were recorded in more than half of the examined individuals (6 patients). The positive dynamics of the increase in the level of γ-IFN and α-IFN in patients of the main group and itsalmost absence in the control group were established.The dynamics of surface expression of the NKG2D receptor on peripheral blood NK cells showed an increase parameters under the influence of the Esberitox drug compared with Valacyclovir monotherapy, with no significant changes in other lymphocyte subpopulations. Conclusions. The administration of Esberitox in case of frequently recurring herpes virus infection is a promising method of combined immuno-adaptive therapy, which requires further careful study.

High-throughput sequencing in diagnostics and prevention of herpes simplex virus infection (Herpesviridae, Alphaherpesvirinae, Simplexvirus, Human alphaherpesvirus 1)

Herpes simplex viruses types 1 (HSV-1) and 2 (HSV-2) are among the most common viruses in the human population. The clinical manifestations of HSV infection vary widely, which necessitates reliable molecular methods for the timely diagnosis of herpes virus infection, as well as for detection of mutations in the genes responsible for drug resistance. PCR is often unable to detect HSV isolates with nucleotide substitutions at the primer binding site. Sanger sequencing of the whole genome reveals mutations mainly at the consensus level, which accumulate at advanced stages of viral infection. High-throughput sequencing (HTS, next generation sequencing) offers an obvious advantage both in early diagnosis of herpes virus infection and identification of HSV variants.