Local Immune Status Research Articles

Skin microbiota analysis in a patient and its role in clinical practice

The skin is structurally a complex barrier organ that forms an original dynamic ecosystem colonized by a wide range of bacteria, fungi, viruses, and mites. The skin microbiome plays an important role in maintaining homeostasis and the state of local immunity and affects pathogenic microorganisms and skin barrier function through bacterial enzyme production. The most frequent and common infectious skin lesions are pyoderma, atopic dermatitis, psoriasis, hidradenitis suppurativa, acne vulgaris, rosacea, seborrheic dermatitis, various mycoses, as well as bacterial and fungal folliculitis. A detailed study of the impact of homeostatic balance on the health of the skin plays an important role in the diagnosis and treatment of skin diseases. Further investigations and deciphering bacteria, fungi, and viruses colonizing the skin will contribute to more effective diagnosis and identification of associations between the microbiota and the occurrence of various diseases, as well as to the development of more effective methods for their treatment, which is of great importance in clinical practice.

State of local immunity of respiratory organs in calves with bronchopneumonia

Respiratory diseases are widespread among cattle and represent a significant proportion of the causes of abandonment in dairy and beef cattle. Despite the fact that new vaccines are being developed and introduced to combat respiratory diseases, which create humoral immunity to the main specific pathogens of respiratory diseases, the problem persists. Thus, it can be assumed that a significant proportion of respiratory diseases is caused by opportunistic microflora. Previous studies have shown that in calves under the influence of adverse microclimate factors, the microflora of the upper respiratory tract migrates to the underlying parts of the respiratory tract, resulting in bronchopneumonia.The study of local immunity indicators in healthy calves and calves with bronchopneumonia was realized. The study involved 8 Holstein calves aged 4-5 months, which were divided into 2 groups. Group 1 (control, n = 3) - clinically healthy calves, group 2 (experiment, n = 5) – calves with bronchopneumonia. Nasal secretions were collected from calves to assess local protection of the respiratory tract. The level of lysozyme, mucin, alkaline phosphatase, total protein, IgM, IgA and IgG was determined in it. The study of nonspecific factors protecting the mucous membrane of the respiratory tract showed a decrease in the level of lysozyme by 27.4% (p ˂ 0.01), as well as an increase in mucin by 76.4% and alkaline phosphatase - by 2.3 times compared with healthy calves. At the same time, with the development of pneumonia, there was an increase in the level of immunoglobulins M and A by 2.5 times (p ˂ 0.05). Thus, the conducted studies have shown that in case of bronchopneumonia, the barrier functions of the respiratory tract are activated to a large extent. In the nasal mucus of sick calves, the level of secretory immunoglobulins A and M increases, the production of alkaline phosphatase and mucin is activated.

Prognostic implications of histologic growth patterns and tumor-infiltrating macrophages in colorectal liver metastases.

Histopathologic patterns at the invasion fronts of tumors predict metastatic potential and prognosis in several cancers. We examined whether such patterns at the interface between colorectal liver metastases and hepatic parenchyma have similar prognostic value. Microscopic growth patterns at edges of metastases including desmoplasia, pushing borders, and replacement of hepatocytes were retrospectively analyzed with respect to surgical outcomes in 142 patients who underwent hepatectomy for colorectal metastases. Patterns included desmoplasia in 58 patients (41%), hepatocyte replacement in 41 (29%), and pushing borders in 43 (30%). Maximum metastasis diameter and serum carcinoembryonic antigen concentration in patients showing desmoplastic tumor growth were lower than those in others (P < 0.05 and P < 0.01). Disease-free survival and overall survival were better in patients showing desmoplastic growth, while a non-desmoplastic tumor growth pattern showed a negative influence. More cluster of differentiation (CD) 68-positive M1 macrophages and fewer CD206-positive M2 macrophages were demonstrated at interfaces of tumors with hepatic parenchyma when desmoplasia was present, although markers for proliferative activity (MIB1 index) and metastatic potential (E-cadherin expression) appeared uninfluenced by desmoplasia. Better long-term results were associated with metastatic tumors showing desmoplastic growth patterns at invasion fronts, which may reflect local immune state in a prognostically useful manner.

INDICATORS OF DENTAL HEALTH AND LOCAL IMMUNITY IN YOUNG ADULTS WHO HAVE SUFFERED FROM CORONAVIRUS INFECTION.

The aim: To determine the dental status and state of local immunity in young adults who have suffered from the coronavirus disease. Materials and methods: The main group consisted of 30 people aged 20-22 years, who suffered from the coronavirus infection Covid19 6.1±1.2 months ago. The comparison group included 20 people who did not have a coronavirus infection. The control group consisted of 35 people, randomized by age and sex, who did not have signs of caries and periodontal tissue disease and did not have coronavirus disease. All patients were examined for dental status and local immunity. Results: The analysis of indicators of dental status revealed the possibility of the existence of a relationship between the signs of acute SARS-Cov2 viral infection and the development of caries and periodontal tissue diseases. Significant changes in the local immunity of the oral cavity were found in the examined patients, which had a pathogenetic influence on the development and progression of caries and periodontal tissue diseases: a significant increase in the level of Ig G, as well as a probable decrease in the concentration of SIg A relative to the comparison group, a probably higher normative value of pathogenic small- and medium-molecular CICs with a significant decrease in the level of physiological large-sized CICs relative to the comparison group, a decrease in the content of anti-inflammatory IL-4, as well as increased concentration of pro-inflammatory cytokines. Conclusions: Young adults who have suffered a coronavirus infection during the last 6 months have significantly higher caries prevalence, bleeding index, PMA index and hygiene index, halitosis, which indicates deeper tissue damage and tooth pathology with the formation of dentition defects than in the comparison group. Indicators of local immunity of the oral fluid have a deep and specific character.

Reduction of Staphylococcus epidermidis in the mammary tumor microbiota induces antitumor immunity and decreases breast cancer aggressiveness

The mammary gland hosts a microbiota, which differs between malignant versus normal tissue. We found that aerosolized antibiotics decrease murine mammary tumor growth and strongly limit lung metastasis. Oral absorbable antibiotics also reduced mammary tumors. In ampicillin-treated nodules, the immune microenvironment consisted of an M1 profile and improved T cell/macrophage infiltration. In these tumors, we noted an under-representation of microbial recognition and complement pathways, supported by TLR2/TLR7 protein and C3-fragment deposition reduction. By 16S rRNA gene profiling, we observed increased Staphylococcus levels in untreated tumors, among which we isolated Staphylococcus epidermidis, which had potent inflammatory activity and increased Tregs. Conversely, oral ampicillin lowered Staphylococcus epidermidis in mammary tumors and expanded bacteria promoting an M1 phenotype and reducing MDSCs and tumor growth. Ampicillin/paclitaxel combination improved the chemotherapeutic efficacy. Notably, an Amp-like signature, based on genes differentially expressed in murine tumors, identified breast cancer patients with better prognosis and high immune infiltration that correlated with a bacteria response signature.This study highlights the significant influence of mammary tumor microbiota on local immune status and the relevance of its treatment with antibiotics, in combination with breast cancer therapies.

Molecular Characterization of Primary and Metastatic Colon Cancer Cells to Identify Therapeutic Targets with Natural Compounds.

Metastasis is the world's leading cause of colon cancer morbidity. Due to its heterogeneity, it has been challenging to understand primary to metastatic colon cancer progression and find a molecular target for colon cancer treatment. The current investigation aimed to characterize the immune and genotypic profiles of primary and metastatic colon cancer cell lines and identify a molecular target for colon cancer treatment. Colony-forming potential, migration and invasion potential, cytokine profiling, miRNA, and mRNA expression were examined. Molecular docking for the Wnt signaling proteins with various plant compounds was performed. Colony formation, migration, and invasion potential were significantly higher in metastatic cells. The primary and metastatic cells' local immune and genetic status revealed TGF β-1, IL-8, MIP-1b, I-TAC, GM-CSF, and MCP-1 were highly expressed in all cancer cells. RANTES, IL-4, IL- 6, IFNγ, and G-CSF were less expressed in cancer cell lines. mRNA expression analysis displayed significant overexpression of proliferation, cell cycle, and oncogenes, whereas apoptosis cascade and tumor suppressor genes were significantly down-regulated in metastatic cells more evidently. Most importantly, the results of molecular docking with dysregulated Wnt signaling proteins shows that peptide AGAP and coronaridine had maximum hydrogen bonds to β-catenin and GSK3β with a better binding affinity. This study emphasized genotypic differences between the primary and metastatic colon cancer cells, delineating the intricate mechanisms to understand the primary to metastatic advancement. The molecular docking aided in understanding the future molecular targets for bioactive- based colon cancer therapeutic interventions.

ОЦЕНКА ЦИТОКИНОВОГО СТАТУСА РОТОВОЙ ЖИДКОСТИ У ДЕТЕЙ С РЕЦИДИВИРУЮЩИМ АФТОЗНЫМ СТОМАТИТОМ

A special place in the pathogenesis of recurrent aphthous stomatitis is occupied by the state of local immunity, which can affect the clinical course and prognosis of chronic diseases of the oral mucosa. The inflammatory process in recurrent aphthous stomatitis is initiated by stimulation of keratinocytes of the oral mucosa by a currently unknown antigen, which leads to stimulation of T-lymphocytes and the release of cytokines and various interleukins.&#x0D; Cytokines are the main nonspecific humoral factor of immunity, providing the initiation and development of an inflammatory response during the development of a protective immune response. A huge number of works are devoted to the study of cytokine status in recurrent aphthous stomatitis, which plays one of the key roles in the pathogenesis of the disease. However, most of the studies were conducted in patients over the age of 18, in addition, the results obtained are contradictory. This determined the purpose of this study – to assess the cytokine status of oral fluid in children with recurrent aphthous stomatitis. The study examined 70 children who were divided into 2 groups. The main group consisted of 45 children aged 7 to 14 years, patients with recurrent aphthous stomatitis, during the relapse of the disease. The main group was divided into 2 subgroups: A – 20 children with the duration of recurrent aphthous stomatitis up to 5 years, group B – 25 children with the duration of recurrent aphthous stomatitis more than 5 years. All patients in the main group had concomitant diseases, in most cases, diseases of the gastrointestinal tract. The control group consisted of 25 practically healthy children without recurrent aphthous stomatitis. In children with recurrent aphthous stomatitis in the oral fluid, a significant increase in the concentration of the anti-inflammatory cytokine TNF-α and a decrease in the concentration of IL-10, an imbalance in the level of IL-2 depending on the duration of the disease, as evidenced by a pronounced inflammatory process of the oral mucosa with epithelial destruction.

Identification of hub genes, signaling pathways and immune infiltration of recurrent spontaneous abortion based on bioinformatics analysis with clinical verification

ObjectiveThe present study was aimed to identify hub genes associated with recurrent spontaneous abortion (RSA) via both bioinformatics analysis and clinical verification, also to evaluate the related pathways and immune infiltration situation of RSA, for exploring its underlying mechanism. Materials and methodsWe screened candidate hub genes associated with RSA via bioinformatic analysis in the microarray datasets GSE22490 downloaded from the Gene Expression Omnibus (GEO) database. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to validate these hub genes. Several kinds of enrichment analysis were carried out to find out the pathways related to RSA. Additionally, CIBERSORT was used for evaluation of local immune Infiltration status of RSA. ResultsThere were 536 differentially expressed genes (DEGs) including 301 upregulated and 235 downregulated genes in RSA group compared with healthy control group. Four hub genes (STAT3, TLR2, TLR4 and CD86) were finally screened out according to the protein–protein interaction (PPI) network analysis, RT-qPCR and Western blotting. Enrichment analysis showed that Toll-like receptor signaling pathway, neutrophil chemotaxis, chemokine signaling pathway and Fc gamma receptor-mediated phagocytosis were strongly associated with RSA. And in immune infiltration analysis, RSA tissue was found containing a higher proportion of monocytes and eosinophils. ConclusionThis study screened out four hub genes and several important pathways changed in the trophoblastic tissue of RSA patients. We also found that monocytes and eosinophils may be involved in RSA. These findings provide theoretical basis for further studies on the mechanisms of RSA.

Local immune state in the women with miscarriage

Functional state of immunity provides maintenance of immunological tolerance for allogeneic fetus, and high level of local protection against antigenic stimulation. Increased functional activity of phagocytic cells at systemic and local levels may cause impairment of complete fetoplacental complex, thus leading to abortion. The purpose of our study was to assess the state of local immunity in women with pregnancy loss.&#x0D; The study involved 174 women in their reproductive age. The first group consisted of 65 patients (37.4%) with a diagnosis of miscarriage, the history of 2 abortions over the period from conception to the 20th week of gestation. The second group included 37 patients (21.2%) with documented recurrent pregnancy loss, with a history of 3 miscarriages. The control group consisted of 72 conditionally healthy women (41.4%) who had 2 or more term pregnancies with the same partner, without a history of obstetric and gynecological complications. Their mean age was 366 years old. Total number and viability of leukocytes, the indices of functional neutrophil activity, their functional reserve and neutrophil stimulation index were determined in cervical mucosal samples over the first phase of the menstrual cycle.&#x0D; In the first phase of menstrual cycle, the women with miscarriage and recurrent pregnancy loss exhibited a statistically significant increase of phagocytosis by the neutrophils from cervical mucus as compared to the group of conditionally healthy women. Intensity of neutrophil phagocytosis in cervical mucus reached higher values in the women with recurrent pregnancy loss more often, compared to the control group. Evaluation of the functional reserve of cervical mucus neutrophils in the subjects with recurrent pregnancy loss and miscarriage showed a trend towards statistically significant differences: this parameter was higher in the patients than in control group. The ability of cervical mucosal neutrophils to produce reactive oxygen species (both spontaneous and induced) did not show statistically significant differences between the patients and controls.&#x0D; Hence, we have observed aberrant functional activity of neutrophilic granulocytes from cervical mucosa in the groups of women with 2 or more abortions, without changing ability of the cells to produce reactive oxygen species. This finding may be explained by prevalence of oxygen-independent mechanisms of intracellular killing, thus suggesting a role of neutrophils for impaired balance of immunological tolerance in pregnant women at the local level.

Torquetenovirus in pregnancy: Correlation with vaginal microbiome, metabolome and pro-inflammatory cytokines.

Torquetenovirus (TTV) is a negative sense, single-stranded DNA virus present in many body fluids of apparently healthy individuals. At present, it is considered a non-pathogenic endogenous virus. TTV can be detected in the vagina of pregnant women, its abundance being modulated with the extent of immune system activation. Until now, there is only scarce information regarding the association between TTV and the composition of the vaginal environment. Therefore, this study aimed to assess the presence of TTV in the vaginal ecosystem of a cohort of white women with a normal pregnancy (n = 60) at different gestational stages (first, second and third trimester) and in 9 subjects suffering a first trimester miscarriage. For each woman, we determined (i) the presence and titer of TTV, (ii) the vaginal bacterial composition by means of Nugent score and 16S rRNA gene sequencing, (iii) the vaginal metabolic profiles through 1H-NMR spectroscopy, and (iv) the vaginal concentration of two pro-inflammatory cytokines (IL-6 and IL-8). More than one third of women were found negative for TTV at all gestational stages. Although not statistically significant, the positivity for TTV dropped from 53.3% in the first to 36.6% in the third trimester. TTV loads varied greatly among vaginal samples, ranging between 2 × 101 and 2 × 105 copies/reaction. No difference in TTV prevalence and loads was observed between women with normal pregnancies and miscarriages. The presence of TTV was more common in women with a higher vaginal leucocyte count (p = 0.02). The levels of IL-6 (p = 0.02), IL-8 (p = 0.03), propionate (p = 0.001) and cadaverine (p = 0.006) were significantly higher in TTV-positive samples. TTV titer was positively correlated with the concentrations of 4-hydroxyphenyllactate (p < 0.0001), isoleucine (p = 0.01) and phenylalanine (p = 0.04). TTV-positive samples were characterized by a higher relative abundance of Sneathia (p = 0.04) and Shuttleworthia (p = 0.0009). In addition, a trend toward a decrease of Lactobacillus crispatus and Lactobacillus jensenii, and an increase of Lactobacillus iners was observed for TTV-positive samples. In conclusion, we found that TTV is quite common in women with normal pregnancy outcomes, representing a possible predictor of local immune status.

A novel 16-gene alternative mRNA splicing signature predicts tumor relapse and indicates immune activity in stage I–III hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is a lethal disease with high relapse and dismal survival rates. Alternative splicing (AS) plays a crucial role in tumor progression. Herein, we aim to integratedly analyze the relapse-associated AS events and construct a signature predicting tumor relapse in stage I–III HCC. Methods: AS events of stage I–III HCC with tumor relapse or long-term relapse-free survival were profiled to identify the relapse-associated AS events. A splicing network was set up to analyze the correlation between the relapse-associated AS events and splicing factors. Cox regression analysis and receiver operating characteristic curve were performed to develop and validate the relapse-predictive AS signature. Single-sample gene set enrichment analysis (ssGSEA) and the ESTIMATE algorithm were used to assess the immune infiltration status of the HCC microenvironment between different risk subgroups. Unsupervised cluster analysis was conducted to assess the relationship between molecular subtypes and local immune status and clinicopathological features. Results: In total, 2441 ASs derived from 1634 mRNA were identified as relapse-associated AS events. By analyzing the proteins involved in the relapse-associated AS events, 1573 proteins with 11590 interactions were included in the protein–protein interaction (PPI) network. In total, 16 splicing factors and 61 relapse-associated AS events with 85 interactions were involved in the splicing network. The relevant genes involved in the PPI network and splicing network were also analyzed by Gene Ontology enrichment analysis. Finally, we established a robust 16-gene AS signature for predicting tumor relapse in stage I–III HCC with considerable AUC values in all of the training cohort, testing cohort, and entire cohort. The ssGSEA and ESTIMATE analyses showed that the AS signature was significantly associated with the immune status of the HCC microenvironment. Moreover, four molecular subgroups with distinguishing tumor relapse modes and local immune status were also revealed. Conclusion: Our study built a novel 16-gene AS signature that robustly predicts tumor relapse and indicates immune activity in stage I–III HCC, which may facilitate the deep mining of the mechanisms associated with tumor relapse and tumor immunity and the development of novel individualized treatment targets for HCC.

Identification of PSMB9 and CXCL13 as Immune-related Diagnostic Markers for Rheumatoid Arthritis by Machine Learning.

Rheumatoid arthritis (RA) is a chronic inflammatory disease that causes significant physical and psychological damage. Although researchers have gained a better understanding of the mechanisms of RA, there are still difficulties in diagnosing and treating RA. We applied a data mining approach based on machine learning algorithms to explore new RA biomarkers and local immune cell status. We extracted six RA synovial microarray datasets from the GEO database and used bioinformatics to obtain differentially expressed genes (DEGs) and associated functional enrichment pathways. In addition, we identified potential RA diagnostic markers by machine learning strategies and validated their diagnostic ability for early RA and established RA, respectively. Next, CIBERSORT and ssGSEA analyses explored alterations in synovium-infiltrating immune cell subpopulations and immune cell functions in the RA synovium. Moreover, we examined the correlation between biomarkers and immune cells to understand their immune-related molecular mechanisms in the pathogenesis of RA. We obtained 373 DEGs (232 upregulated and 141 downregulated genes) between RA and healthy controls. Enrichment analysis revealed a robust correlation between RA and immune response. Comprehensive analysis indicated PSMB9, CXCL13, and LRRC15 were possible potential markers. PSMB9 (AUC: 0.908, 95% CI: 0.853-0.954) and CXCL13 (AUC: 0.890, 95% CI: 0.836-0.937) also showed great diagnostic ability in validation dataset. Infiltrations of 16 kinds of the immune cell were changed, with macrophages being the predominant infiltrating cell type. Most proinflammatory pathways in immune cell function were activated in RA. The correlation analysis found the strongest positive correlation between CXCL13 and plasma cells, PSMB9, and macrophage M1. There is a robust correlation between RA and local immune response. The immune-related CXCL13 and PSMB9 were identified as potential diagnostic markers for RA based on a machine learning approach. Further in-depth exploration of the target genes and associated immune cells can deepen the understanding of RA pathophysiological processes and provide new insights into diagnosing and treating RA.

Modern ideas about the etiology, pathogenesis, diagnosis and treatment of chronic nonspecific cervicitis

Currently, cervicitis of nonspecific etiology is one of the urgent problems of modern gynecology due to the extremely high frequency of their occurrence, a tendency to a chronic relapsing course, a negative impact on reproductive health, and the risk of developing a number of complications. The role of violations of vaginal microbiocenosis and local immune status in the development of CNC has been convincingly shown. The replacement of lactobacilli with predominantly anaerobic microorganisms (Ureaplasma, Mycoplasma, Gardnerella vaginalis, Prevotella, Peptostreptococcus spp. and Bacteroides spp.), characteristic of bacterial vaginosis, is extremely often detected in patients with CNC. Of particular importance from the point of view of clarifying the pathophysiological mechanisms of development and the development of new diagnostic and prognostic markers, as well as the personalization of CNC therapy, is the study of the cytokine status. Cervicitis and other inflammatory diseases of the lower genital tract are characterized by an increase in the expression of pro-inflammatory cytokines with simultaneous inhibition of the formation of anti-inflammatory cytokines. Significant multidirectional changes in the balance of pro- and anti-inflammatory cytokines were revealed in active and chronic sluggish cervicitis. It should be noted that, despite the variety of proposed approaches to CNC therapy, this problem is still far from being solved to date. One of the promising directions is the personalization of CNC therapy based on molecular genetic markers (polymorphisms of cytokine genes, Toll-like receptors, genes of the detoxification system). The use of such a personalized approach can significantly increase the effectiveness of CNC treatment and reduce the risk of disease recurrence.

Clinical and immunological characteristics of pediatric patients with stomatitis

The aim of study. To study the local immune status in pediatric patients diagnosed with stomatitis. Objects and methods. The work is based on a retrospective and prospective analyses result research among 120 pediatric patients (4-12 years old) with stomatitis. Results. The results of patients' immunological examination with stomatitis are described. Research data (clinical, immunological) were analyzed using STATISTICA 10.0 software. In patients of the primary physical and histological period, the concentrations of α-amylase, lysozyme, myeloperoxidase, and secretory immunoglobulin A are increased. There is a lack of local immunity and production of immunoglobulin M. In pediatric patients related to the secondary period of physical and histological age features, the production of lysozyme, lactoferrin and myeloperoxidase, as well as immunoglobulin M and secretory immunoglobulin A, is reduced. Conclusion. From the results of this research, it is evident that there is a correlation between the physico-histological and immunological characteristics in children diagnosed with stomatitis.

Local cellular immunity in colorectal cancer depending on tumor site.

e15613 Background: Colorectal cancer (CRC) is the third most commonly diagnosed cancer. More than 60% of all cases of CRC are colon cancer (CC). The role of individual units of the immune system in the development of this tumor is ambiguous. The purpose of this study was to characterize local populations and subpopulations of immunocompetent cells in colorectal cancer with different tumor locations. Methods: The study included 50 CC (adenocarcinoma) patients aged 35-86 years, women n = 26 (52%). Stage I tumors were registered in 4 patients (8%), stage II 25 (50%), stage III 21 (42%). 20 patients (40%) had right-sided CC (group 1), 9 (18%) left-sided CC (group 2), and 21 (42%) sigmoid CC (group 3). All patients received surgical treatment. Cell suspensions were obtained from tumor (TT) and peritumoral tissues (PT) (1-3 cm from the tumor), and then processed with an antibody panel in accordance with the manufacturer instructions (Becton Dickinson, USA) to identify the main populations and subpopulations of leukocytes and lymphocytes. The relative number of major populations and subpopulations of lymphocytes was determined on the BD FACSCanto flow cytometer. Results: A decrease in lymphocytic infiltration was noted left-sided tumors and sigmoid tumors compared to right-sided CC, by 46% and 51%, respectively. The percentage of CD3+ cells was almost the same regardless of the colon tumor location, and the number of main populations of T lymphocytes differed: in group 3, the content of CD4+ cells was 21% higher, and CD8+ cells were 22% lower compared with group 1, while group 2 had no differences. Group 2 differed in the tumor infiltration with both NK and NKT lymphocytes, which were higher by 25% and 22%, respectively, than in group 1. An increase in NK cells was noted in the sigmoid colon (group 3), and the relative number of NKT lymphocytes decreased. A common feature of TT in groups 2 and 3 was an increase in the content of B lymphocytes by 98% and 133%, respectively. PT of group 2 showed a decrease by 72%, 33%, 66%, and 46% in the relative number of lymphocytes, CD4+, NKT and B lymphocytes compared with group 1, together with an increased content of NK and CD8+ lymphocytes. PT in patients of group 3 had a decrease in the number of total and NK lymphocytes by 46% and 26%, respectively, and a significant increase by 85% in the content of CD8+ cells, with no changes in CD3+, CD4+ cells, B and NKT lymphocytes. Conclusions: The local immune status of CRC patients demonstrated a number of differences: right-sided tumors were characterized by a higher T-lymphocytic infiltration with the same tendency in the perifocal tissues, while left-sided and sigmoid tumors showed higher B-lymphocytic infiltration, which can help in the disease prognosis and choice of treatment strategy.

Circulating extracellular vesicles carrying Firmicutes reflective of the local immune status may predict clinical response to pembrolizumab in urothelial carcinoma patients.

Bacterial flora has clinical significance for the host. The metabolic environment created by this flora influences immunotherapy in urothelial carcinoma. However, there are no reports on the clinical significance of bacterial flora in the host bloodstream. We aimed to clarify the correlation between extracellular vesicle (EV)-derived blood microflora information and tumor immunological status in urothelial carcinoma (UC) patients. Serum samples were collected from 20 healthy donors, 50 patients with localized UC, and 31 patients with metastatic UC (mUC) who had undergone pembrolizumab treatment. Bacterial DNA in EVs was extracted from each sample. Metagenomic sequencing was performed after amplification of the V1-V2 region of the bacterial 16S rRNA gene. Using the matched tumor tissue and serum samples, we revealed that the smaller amount of peripheral EVs carrying Firmicutes DNA was significantly correlated with the higher number of infiltrating T cells within tumor tissues (CD3; p = 0.015, CD4; p = 0.039, CD8; p = 0.0084) and the higher expression of activation markers on their surface (ICOS on both CD4; p = 0.0013 and CD8 T cells; p = 0.016 and 4-1BB on CD4 T cells; p = 0.016). In terms of circulating metabolic information, L-Ser and L-Pro levels, which play important roles in T cell expansion and proliferation, were significantly higher in the Firmicutes-low group (p = 0.010). All of the patients with higher Firmicutes abundance had disease progression without any clinical response (p = 0.026) and significantly inferior prognosis for pembrolizumab therapy (p = 0.035). This is the first study on the importance of peripheral bacterial EVs in cancer patients treated with cancer immunotherapy.

Upregulation of FOXP3+ Regulatory T Lymphocytes and CD8+ Lymphocytes in Patients with High-Grade Squamous Intraepithelial Lesions Correlated with HPV Infection

Abstract Modern therapeutic strategies for precancerous cervical intraepithelial neoplasia (CIN) focus on immune-modulatory and cancer vaccination. The local cervical immune status in cervical cancer and CIN could influence HPV infection persistence, progression and carcinogenesis. We analysed the role of FOXP3+ regulatory T lymphocytes, CD4+ and CD8+ T lymphocytes in CIN I, CIN II and CIN III patients with and without HPV infection. Sixty-two patients were enrolled in the study. Each patient underwent a colposcopy-guided cervical biopsy. FOXP3+ lymphocytes and CD4+,CD8+ lymphocytes were detected by immunostaining in tissue samples obtained from a control group (n = 10), patients with CIN I (n = 20), CIN II (n = 14) and CIN III (n = 18) lesions. HPV was assayed by Aptima. The results showed that the numbers of CD4+ T lymphocytes did not differ between patients with CIN I, CIN II, and CIN III. However, patients with CIN II and CIN III had significantly upregulated CD8+T lymphocytes compared to patients with CIN I. In addition, patients with CIN II and CIN III had increased FOXP3 + T lymphocytes compared with patients with CIN I, which was associated with HPV status. Upregulation of FOXP3+ regulatory T lymphocytes and CD8-positive lymphocytes in patients with CIN II and CIN III suggested a pivotal role of T regulatory lymphocytes and CD8+ lymphocytes for counteracting the host immune response in the progression from CIN I to CIN II and CIN III.

Стан мікрофлори та ступінь вираженості місцевого імунного захисту в ендометрії жінок з безпліддям, що розвивається на тлі хронічного ендометриту поєднаного з бактеріальним вагінозом

The purpose of the study was to evaluate the taxonomic profile of the microbiota and the state of local immunity in the endometrium of women with infertility under conditions of chronic endometritis combined with bacterial vaginosis. Materials and methods. Diagnosis of chronic form of endometritis was performed on clinical symptoms, medical history, scraping, endoscopic and ultrasound examinations. All women were divided into three groups: Group I – 12 healthy women, who had no signs of chronic endometritis and formed a control group; Group II – patients with severe chronic endometritis (n = 16); Group III – patients with chronic endometritis developed on the background of bacterial vaginosis (n = 12). Diagnosis of bacterial vaginosis in patients was performed by detecting an increase in the pH of vaginal secretions &gt;4.5, the presence of homogeneous vaginal discharge with an unpleasant odor, detection of vaginal epitheliocytes with adherent gram-variable microorganisms, detection of "fishy" odor. The study was performed in the second phase of the menstrual cycle (day 18-23). Endometrial aspirate for the study of microflora was selected using a device for obtaining endometrial cytology Endobrash Standard for Endometrial Cytology. Results and discussion. It was found that the development of chronic endometritis on the background of bacterial vaginosis increases the frequency of detection of opportunistic anaerobic microflora in the endometrium compared to women without bacterial vaginosis. It has been shown that a decrease in the level of T-helpers and an increase in the level of T-suppressors / killers in the endometrium are exacerbated in patients with chronic endometritis developed against the background of bacterial vaginosis. Thus, the level of CD4 + cells decreased by 1.6 times, and the level of CD8 + cells increased by 1.9 times compared to healthy women. Against this background, the level of B-lymphocytes and IgA in the endometrium of women with chronic endometritis and bacterial vaginosis decreased. Conclusion. The development of chronic endometritis is associated with an increase in the frequency of detection and quantitative content in the endometrium of Streptococcus spp., Staphylococcus spp. The development of bacterial vaginosis is accompanied by the detection in the endometrium of new species of microorganisms – Prevotella spp., Mobiluncus spp. The imbalance of the endometrial microflora is accompanied by significant violations of local cellular and humoral immune defense, which is expressed by a decrease in the level of T-helpers, B-lymphocytes and IgA with a simultaneous increase in T-suppressors / killers in women with chronic endometritis. The established changes amplify at a bacterial vaginosis

Protective immune trajectories in early viral containment of non-pneumonic SARS-CoV-2 infection

The antiviral immune response to SARS-CoV-2 infection can limit viral spread and prevent development of pneumonic COVID-19. However, the protective immunological response associated with successful viral containment in the upper airways remains unclear. Here, we combine a multi-omics approach with longitudinal sampling to reveal temporally resolved protective immune signatures in non-pneumonic and ambulatory SARS-CoV-2 infected patients and associate specific immune trajectories with upper airway viral containment. We see a distinct systemic rather than local immune state associated with viral containment, characterized by interferon stimulated gene (ISG) upregulation across circulating immune cell subsets in non-pneumonic SARS-CoV2 infection. We report reduced cytotoxic potential of Natural Killer (NK) and T cells, and an immune-modulatory monocyte phenotype associated with protective immunity in COVID-19. Together, we show protective immune trajectories in SARS-CoV2 infection, which have important implications for patient prognosis and the development of immunomodulatory therapies.

Relationship between serum lipid levels and the immune microenvironment in breast cancer patients: a retrospective study

BackgroundTherapeutic agents for dyslipidaemia, in particular statins, have been recently reported to suppress growth and metastasis of breast cancer. However, the predictive value of lipid control in breast cancer patients has not been discussed sufficiently. In addition, though immunometabolism is a relatively novel approach for tumour immunotherapy, the relationship between lipid metabolism and immune status has not been well documented. We therefore investigated the effects of lipid metabolism on antitumour immune response and cancer prognosis.MethodsExcept for patients with ductal carcinoma in situ, 938 patients treated with curative surgery were examined. The correlation between treatment for dyslipidaemia or serum lipid levels and clinicopathological features, including the prognosis, was evaluated retrospectively. Also, we stratified these results by intrinsic subtype of breast cancer, menopause, and type of therapeutic agents for dyslipidaemia. Moreover, neutrophil- to-lymphocyte ratio (NLR) and tumour-infiltrating lymphocytes (TILs) were used as indicators of systemic and local immune status, respectively.ResultsOf 194 patients treated for dyslipidaemia, recurrence-free survival (RFS) and overall survival (OS) did not differ significantly between users of drugs for dyslipidaemia and non-users (p = 0.775 and p = 0.304, log-rank, respectively). Among postmenopausal, hormone receptor (HR)-positive/human epidermal growth factor receptor 2 (HER2)-negative breast cancer patients treated for dyslipidaemia, the good serum lipid control group had significantly better RFS (p = 0.014, log-rank), lower postoperative NLR (p = 0.012), and higher TILs in resected tissues (p = 0.024) than the poor control group. Multivariate analysis showed that postoperative serum lipid levels were a risk factor for recurrence (hazard ratio = 4.722, 95% confidence interval 1.006–22.161, p = 0.049).ConclusionsGood control of serum lipid metabolism may improve the tumour immune microenvironment and prognosis in postmenopausal HR-positive/HER2-negative breast cancer patients.