The identification of risk factors for early pregnancy termination is an important aspect in modern obstetrics. The vaginal microbiota plays a significant role in normal pregnancy course affecting the local immune response. Inflammation caused by pathogenic and opportunistic microorganisms activates various immune arms altering expression of innate immunity genes. Molecular genetic determination of the inflammatory response and microbial composition of the vaginal biotope could become an important criterion for predicting pregnancy complications. The aim of the study was to analyze markers of the local immune response and vaginal microbiota composition during pregnancy to predict pregnancy outcomes. Materials and methods. 52 women verified at up to 10 weeks pregnancy were examined. Depending on the outcome of ongoing pregnancy, study participants were divided into 3 groups: group 1 (n = 42) with term birth; group 2 (n = 7) with premature birth; and group 3 (n = 3) with miscarriage. Vaginal discharge was analyzed using a molecular genetic test to evaluate local cytokines and vaginal microbiota profile, sequentially in each trimester. Results. In all patients during the first trimester, the presence of lactobacillus species dominated by L. iners and L. crispatus was detected. L. crispatus prevailed only in group 1 in the first and second trimesters. In group 2, L. iners was most common in both the first and second trimesters, with a significantly higher level in the second trimester compared to group 1. The inflammation index in the first trimester was more often associated with inflammation in groups 1 and 2. In the second trimester, the cytokine profile in group 1 showed no signs of inflammatory response, while in group 2, pro-inflammatory responses were significantly more common. Conclusion. The dominance of L. crispatus in the vaginal microbiota and the low expression of pro-inflammatory cytokines during the second trimester may suggest a high likelihood for successful pregnancy outcome. Assessing cytokine profile along with type of vaginal microbiocenosis opens up avenues for diagnosis allowing to identify women at risk of premature birth.
As part of the 365-day isolation experiment “SIRIUS-23” a relationship between hormone concentrations (estradiol and prolactin) and vaginal microbiota composition in four female participants was assessed. Vaginal swabs were collected 10–14 days before the start of isolation, on days 36, 64, 102, 123, 186, 256, and 339 of isolation study, and 10 days after its cessation. Blood estradiol and prolactin concentrations were measured using an enzyme-linked immunosorbent assay, while microbiological analysis included microbial culture on selective media and identification using MALDI-ToF MS, followed by statistical data processing using factorial and regression analysis, and nonparametric tests (Friedman, Mann–Whitney). The study revealed that estradiol concentrations decreased after the onset of isolation, reaching a minimum level on day 123 (p = 0.05), while prolactin demonstrated an upward trend on day 36, followed by a statistically significant decline on day 64. Factor analysis identified three main factors influencing the “hormone-bacteria” system: i) combined opportunistic microorganisms (OM) and prolactin; ii) included estradiol concentration and its antagonistic relation with some OM number; iii) mirrored competitive interplay between microorganisms. Estradiol concentration correlated positively with protective lactobacilli count (L. aginalis, L. jensenii, L. crispatus) and negatively with OM count, while prolactin concentration was more often positively associated with OM count (S. hominis, S. anginosus, C. amycolatum) and negatively with lactobacilli count. Regression analysis confirmed direct and inverse relationships, such as a positive association between prolactin concentration and E. coli count on day 64 and an inverse association between estradiol concentration and S. anginosus count. Stress factors (resource limitation, psychoemotional stress) exacerbated microbiota imbalance through decreased estradiol and increased prolactin level, which was particularly evident on day 102 of isolation, when OM count peaked in parallel with decreased lactobacilli count. The results highlight the key role for estradiol in maintaining vaginal microbiota balance and the opposite effect of prolactin associated with OM, which justifies a need to monitor hormonal levels and microbiota in women under extreme conditions (e.g., during space flights) and development of personalized probiotic strategies to minimize dysbiosis risks.
The Ob-Irtyshsky focus, which includes the Tyumen region is largest among Opisthorchis felineus invasion sites. The high intensity of invasion spreading in the outbreak underlies a high risk of infection often occurring with acute stage for the residents of the Tyumen region. The aim of this study was to evaluate the immunological parameters in patients with acute opisthorchiasis compared to healthy subjects. Ten patients at the clinic of the Tyumen Research Institute of Regional Infectious Pathology were examined with a diagnosis of acute opisthorchiasis. Immunological examination included lymphocyte phenotyping by flow cytometry. The concentration of immunoglobulins M, G, A, and E and cytokines IFNγ, IL-4, IL-8, IL-10 in blood serum was quantitated by enzyme immunoassay. Neutrophil absorbance was investigated in latex particle test (d = 1 μm). Neutrophil myeloperoxidase enzyme activity was measured spectrophotometrically, concentration of circulating immune complexes — by precipitation with PEG-6000 solution. In patients with acute opisthorchiasis, increased count of leukocytes, eosinophils, monocytes was detected; increased neutrophil scavenging and enzymatic activity; increased absolute number of CD3+ cells, absolute number of T-lymphocytes with CD3+HLA-DR+ marker, relative and absolute number of CD4+ cells were found as well. The concentration of total immunoglobulins G, A, M as well as circulating immune complexes and pro-inflammatory cytokine IL-8 were increased. A positive correlation between IL-8 levels and count of eosinophils, monocytes and percentage of natural killers was revealed in patients but not in control group. Patients have decreased levels of IFNγ and the cytokine IL-10. The level of IL-4 did not differ between patients with opisthorchiasis vs. control subjects. The correlation between IL-4 level and circulating immune complexes, count of eosinophils and natural killer cells was found in patients but not in control group. Thus, patients with acute opisthorchiasis were characterized by activated innate immune response as well as cellular and humoral arms of the adaptive immune response.
Laboratory data are important for confirming diagnosis, assessing COVID-19 progression, and determining treatment measures. The aim of this study was to evaluate the associations of hematological, coagulation parameters, and systemic inflammatory markers with COVID-19 severity according to computed tomography data in hospitalized patients. Materials and methods. Clinical blood test parameters were assessed in 356 patients with COVID-19. Results. Patients with severe vs. minimal and moderate lung tissue damage have a significant increase in the absolute leukocyte count, the relative and absolute number of neutrophils, and a decrease in the relative number of monocytes and lymphocytes. These differences are also observed in patients with moderate vs. minimal lung tissue damage, except for the absolute leukocyte count. However, the latter as well as the relative number of neutrophils and lymphocytes in patients with minimal and moderate lung tissue damage, along with the absolute number of neutrophils and the relative number of monocytes in all patients were within the reference range. In COVID-19 patients with severe lung tissue damage, markers of systemic inflammation (neutrophil-lymphocytic index, platelet-lymphocytic index, systemic inflammatory response index, systemic inflammation index and cumulative systemic inflammation index) were higher than in those with the minimum and average volume, and the monocyte-lymphocyte index is higher than in those with the average volume of lung tissue damage. At the same time, in patients with minimal lesion volume, neutrophil-lymphocyte index and systemic inflammation index were at lower level, than in case of average volume of lung tissue lesion. Conclusion. The study revealed an association between the relative count of neutrophils and lymphocytes, neutrophil-lymphocyte ratio, systemic inflammation index, and systemic inflammatory response index, aggregate systemic inflammation index and severe COVID-19 disease based on computed tomography data. These biomarkers provide valuable information about systemic inflammation and immune system activity and can be detected using standard blood tests.
Human herpesviruses 6A and 6B (HHV6A, HHV6B) are ubiquitous pathogens with lifelong persistence in the human body. HHV6A/B infection often accompanies HIV infection, however, data on the interaction between the two viruses are contradictory. The discrepancy in study results on the influence of HHV6A/B on the course of HIV infection may be due to the molecular genetic variability of the betaherpesviruses. The aim of this work was to assess the molecular genetic diversity of HHV6A and HHV6B in HIV-infected adults in the Nizhny Novgorod region. Materials and methods. Blood leukocyte isolates from 138 HIV-infected patients (HIV(+)) and 68 HIV-uninfected (HIV(–)) individuals were studied. HHV6A/B species were identified by PCR with agarose gel electrophoresis. Nucleotide sequences of the U90A (206 bp) and U90B (431 bp) gene fragments were obtained by Sanger sequencing. The intra-species genovariant of the examined betaherpesviruses was determined based on signature nucleotide substitutions of the sequenced sequences and in accordance with the author’s classification. Results. HHV6A/B DNA was detected in 39% of HIV(+) and 46% of HIV(–) examined individuals. HHV6A (genovariant GV3a) was identified in one HIV(+) sample. The remaining samples from both groups contained HHV6B. In the group of HIV-infected patients, dominant genovariants GV1a, GV2b, and GV2e (~30% each) were evenly distributed, and the minor GV2a (7.3%) was also detected. In the group of clinically healthy virus carriers, genovariant GV2e predominated (72.4%), while GV1a and GV2b were less common. The likelihood of detecting GV2e decreased (OR = 0.18 (0.04; 0.76), p = 0.031), whereas that of for GV1a and GV2b variants increased (in both cases OR = 5.48 (1.31; 28.52), p = 0.016) in HIV(+) individuals compared to control group. Analyzing age structure for HHV6B genovariants showed that in HIV-infected patients, the diversity of genovariants declines with decreasing patient age. Сonclusion. The obtained results indicate the need to investigate the molecular genetic diversity of HHV6A and HHV6B for effective diagnosis and interpretation of findings in HIV-infected patients.
