Serum Cytokine Alterations Associated with Age of Patients with Nephropathia Epidemica

Nephropathia epidemica (NE) is a zoonotic disease caused by hantaviruses transmitted from rodents, endemic in the Republic of Tatarstan, Russia. The disease presents clinically with mild, moderate, and severe forms, and time-dependent febrile, oliguric, and polyuric stages of the disease are also recognized. The patient’s cytokine responses have been suggested to play a central role in disease pathogenesis; however, little is known about the different patterns of cytokine expression in NE in cohorts of different ages and sexes. Serum samples and clinical records were collected from 139 patients and 57 controls (healthy donors) and were used to analyze 48 analytes with the Bio-Plex multiplex magnetic bead-based antibody detection kits. Principal component analysis of 137 patient and 55 controls (for which there was full data) identified two components that individually accounted for >15% of the total variance in results and together for 38% of the total variance. PC1 represented a proinflammatory TH17/TH2 cell antiviral cytokine profile and PC2 a more antiviral cytokine profile with patients tending to display one or the other of these. Severity of disease and stage of illness did not show any correlation with PC1 profiles; however, significant differences were seen in patients with high PC1 profiles vs. lower for a number of individual clinical parameters: High PC1 patients showed a reduced number of febrile days, but higher maximum urine output, higher creatinine levels, and lower platelet levels. Overall, the results of this study point towards a stronger proinflammatory profile occurring in younger NE patients, this being associated with markers of acute kidney injury and low levels of high-density cholesterol. This is consistent with previous work indicating that the pathology of NE is immune driven, with an inflammatory immune response being associated with disease and that this immune response is more extreme in younger patients.

Diphtheria toxin induced but not CSF1R inhibitor mediated microglia ablation model leads to the loss of CSF/ventricular spaces in vivo that is independent of cytokine upregulation

Background Two recently developed novel rodent models have been reported to ablate microglia, either by genetically targeting microglia (via Cx3cr1-creER: iDTR + Dtx) or through pharmacologically targeting the CSF1R receptor with its inhibitor (PLX5622). Both models have been widely used in recent years to define essential functions of microglia and have led to high impact studies that have moved the field forward. Methods Using either Cx3cr1-iDTR mice in combination with Dtx or via the PLX5622 diet to pharmacologically ablate microglia, we compared the two models via MRI and histology to study the general anatomy of the brain and the CSF/ventricular systems. Additionally, we analyzed the cytokine profile in both microglia ablation models. Results We discovered that the genetic ablation (Cx3cr1-iDTR + Dtx), but not the pharmacological microglia ablation (PLX5622), displays a surprisingly rapid pathological condition in the brain represented by loss of CSF/ventricles without brain parenchymal swelling. This phenotype was observed both in MRI and histological analysis. To our surprise, we discovered that the iDTR allele alone leads to the loss of CSF/ventricles phenotype following diphtheria toxin (Dtx) treatment independent of cre expression. To examine the underlying mechanism for the loss of CSF in the Cx3cr1-iDTR ablation and iDTR models, we additionally investigated the cytokine profile in the Cx3cr1-iDTR + Dtx, iDTR + Dtx and the PLX models. We found increases of multiple cytokines in the Cx3cr1-iDTR + Dtx but not in the pharmacological ablation model nor the iDTR + Dtx mouse brains at the time of CSF loss (3 days after the first Dtx injection). This result suggests that the upregulation of cytokines is not the cause of the loss of CSF, which is supported by our data indicating that brain parenchyma swelling, or edema are not observed in the Cx3cr1-iDTR + Dtx microglia ablation model. Additionally, pharmacological inhibition of the KC/CXCR2 pathway (the most upregulated cytokine in the Cx3cr1-iDTR + Dtx model) did not resolve the CSF/ventricular loss phenotype in the genetic microglia ablation model. Instead, both the Cx3cr1-iDTR + Dtx ablation and iDTR + Dtx models showed increased activated IBA1 + cells in the choroid plexus (CP), suggesting that CP-related pathology might be the contributing factor for the observed CSF/ventricular shrinkage phenotype. Conclusions Our data, for the first time, reveal a robust and global CSF/ventricular space shrinkage pathology in the Cx3cr1-iDTR genetic ablation model caused by iDTR allele, but not in the PLX5622 ablation model, and suggest that this pathology is not due to brain edema formation but to CP related pathology. Given the wide utilization of the iDTR allele and the Cx3cr1-iDTR model, it is crucial to fully characterize this pathology to understand the underlying causal mechanisms. Specifically, caution is needed when utilizing this model to interpret subtle neurologic functional changes that are thought to be mediated by microglia but could, instead, be due to CSF/ventricular loss in the genetic ablation model.

Elevated IgA and IL-10 levels in very-early-onset inflammatory bowel disease secondary to IL-10 receptor deficiency

ABSTRACT Objective: To report two patients with very-early-onset inflammatory bowel disease (VEOIBD) secondary to interleukin-10 receptor (IL-10R) mutations, explore immunophenotyping data and plasma cytokine profile on these cases compared to healthy controls, and describe the phenotype of IL-10/IL-10R mutations based on a literature review. Case description: We report on two female infants referred to our tertiary center at the age of ten months, with severe colonic and perianal disease, as well as significant malnutrition, who had shown limited response to usual inflammatory bowel disease (IBD) therapy agents. In the first case, whole-exome sequencing (WES) revealed a homozygous (c.537G>A/p.T179T) mutation in exon 4 of the IL-10RA gene, while in the second patient, compound heterozygosity was identified, also in the IL-10RA gene (chr11:117.859.199 variant A>G/p.Tyr57Cys and chr11: 117.860.335 variant G>T/p.Val123Leu). Both patients underwent hematopoietic cell transplantation (HCT). Immunological work-up of these patients revealed increased IL-10 plasma levels and increased IgA. Comments: Our case reports disclose novel findings on plasma cytokine profile in IL-10R deficiency, and we describe the severe phenotype of IL-10/IL-10R deficiency that should be recognized by physicians.

A Fiber-Rich Diet and Radiation-Induced Injury in the Murine Intestinal Mucosa

Dietary fiber is considered a strong intestinal protector, but we do not know whether dietary fiber protects against the long-lasting mucosal damage caused by ionizing radiation. To evaluate whether a fiber-rich diet can ameliorate the long-lasting pathophysiological hallmarks of the irradiated mucosa, C57BL/6J mice on a fiber-rich bioprocessed oat bran diet or a fiber-free diet received 32 Gray in four fractions to the distal colorectum using a linear accelerator and continued on the diets for one, six or 18 weeks. We quantified degenerating crypts, crypt fission, cell proliferation, crypt survival, macrophage density and bacterial infiltration. Crypt loss through crypt degeneration only occurred in the irradiated mice. Initially, it was most frequent in the fiber-deprived group but declined to levels similar to the fiber-consuming group by 18 weeks. The fiber-consuming group had a fast response to irradiation, with crypt fission for growth or healing peaking already at one week post-irradiation, while crypt fission in the fiber-deprived group peaked at six weeks. A fiber-rich diet allowed for a more intense crypt cell proliferation, but the recovery of crypts was eventually lost by 18 weeks. Bacterial infiltration was a late phenomenon, evident in the fiber-deprived animals and intensified manyfold after irradiation. Bacterial infiltration also coincided with a specific pro-inflammatory serum cytokine profile. In contrast, mice on a fiber-rich diet were completely protected from irradiation-induced bacterial infiltration and exhibited a similar serum cytokine profile as sham-irradiated mice on a fiber-rich diet. Our findings provide ample evidence that dietary fiber consumption modifies the onset, timing and intensity of radiation-induced pathophysiological processes in the intestinal mucosa. However, we need more knowledge, not least from clinical studies, before this finding can be introduced to a new and refined clinical practice.

Gender features of the cytokine profile in patients with chronic atrophic gastritis

Background. Сhronic atrophic gastritis certainly remains an urgent problem of gastroenterology but data on sexual differences in the content of cytokines in this pathology are quite contradictory. The purpose of the study: to assess the gender chara­cteristics of the cytokine profile in patients with chronic atrophic gastritis. Materials and methods. The study included 120 patients with gastric atrophy, according to histological examination of biop­sies. The control group consisted of 20 healthy individuals, men and women equally. In all patients, we have evaluated the levels of interleukins (IL-8, IL-10, IL-18), tumor necrosis factor alpha (TNF-α), vasculoendothelial growth factor (VEGF) by enzyme-linked immunosorbent assay using appropriate reagent from Vector-BEST kits and the Stat Fax 303 Plus analyzer. Results. In men with chronic atrophic gastritis, there is a more pronounced imba­lance towards pro-inflammatory cytokines, in particular the level of IL-18 is 1.7 times higher (p < 0.05) than in women. In 46.2 % of cases, the content of IL-8 was also elevated in men by 1.3 times (p > 0.05) compared to women. The level of anti-inflammatory cytokine IL-10 does not have a significant gender difference in patients with precancerous conditions of the stomach. The median of IL-18/IL-10 ratio in men is 2 times higher than in women: 65.36 (21.67; 154.25) vs. 32.15 (12.76; 191.85) (p < 0. 05). In males, IL-8/IL-10 ratio is also 1.5 times higher, which is 2.25 (1.29; 7.68) vs. 1.49 (0.75; 9.78) but this difference was not statically significant. Serum content of VEGF in men exceeded the same indicator in women by 1.4 times (p < 0.05). Direct correlation between VEGF content and the levels of TNF-α (r = 0.47, p < 0.05), IL-8 (r = 0.42, p < 0.05), IL-18 (r = 0.58, p < 0.05) confirm the evidence of increased VEGF expression under the influence of many proangiogenic growth factors and proinflammatory cytokines. Conclusions. With an increase in the level of IL-18 and VEGF by more than 30 %, men require dynamic monitoring for early detection of precancerous structural changes in the gastric mucosa.

Mononuclear subsets and cytokine profile of venous and capillary blood in patients with psoriasis and healthy people

Psoriasis is considered an autoimmune disease with a predominantly cellular mechanism for the development of disorder. Studies in immune pathogenesis of psoriasis were performed either in animal model, which is not just similar to humans, or the data were obtained in patients by means of skin window method, which is traumatic, or by examining venous blood. However, it is difficult to discern parameters of the local immune response in venous blood samples. We have attempted to find an adequate method which would be convenient both for the patient and for the researcher, in order to assess local immune processes occurring in the skin affected by psoriasis. We examined 20 patients with a verified diagnosis of psoriasis, the average age was 44.3 years. The control group included 15 healthy adults, with average age of 46.6 years. Capillary blood was taken by fingerprick, whereas, in psoriatic patients, the samples were taken near the psoriatic lesion at a final volume of 400 μL in two microvettes. Venous blood (3 mL) was taken from the cubital vein into a vacuum tube with EDTA. Clinical analysis of venous and capillary blood was performed in automated hematological analyzer. Immunophenotyping was performed by four-color staining of whole capillary and venous blood followed by lysis of erythrocytes. Cytofluorometry was performed using techniques and reagents from BD Biosciences (USA). Plasma cytokines were determined by multiplex approach (MagPix, BioRad, USA). Upon clinical analysis of blood, the difference between capillary and venous blood was not found, either in healthy group, or among patients with psoriasis. In healthy people, the subsets of mononuclear cells, did not differ between venous and capillary blood. The samples of capillary and venous blood in the patients with psoriasis showed significantly increased levels of double-positive lymphocytes (CD45RA+/CD45R0+), B lymphocytes and NKT lymphocytes (both for relative and and absolute values). A significant increase in the percentage of naive T lymphocytes, activated helpers (Thact) and Treg, as well as B1 cells and Breg, and a significant decrease in B2 lymphocytes was registered in capillary blood of the patients with psoriasis. In venous blood samples from psoriatic patients, only a significant increase in Thact, Treg, and Breg was revealed. In the capillary blood of patients with psoriasis, we found a significant increase in the levels of non-classical M2 monocytes and inflammatory Minfl monocytes, and a decrease in classical M1 monocyte levels; in venous blood of psoriatic patients, only an increase in inflammatory Minfl monocytes was revealed. In capillary blood, all the studied cytokines in psoriasis patients significantly exceeded the levels of corresponding cytokines in healthy controls, except of IL-10. The levels of this cytokine did not differ from healthy group. In venous blood, the levels of most studied cytokines in the group of patients with psoriasis did not differ from the group of healthy ones. Approximately two-fold increase was revealed for IL-4, IL-21, IL-23 and TNF. First, the subsets of mononuclear cells and the cytokine profile of capillary and venous blood of healthy people did not differ significantly. Secondly, our proposed method for determining the subsets of mononuclear cells and capillary blood cytokines profile from the area of psoriatic lesions may be used to monitor local immunity in the patients with psoriasis. This approach is significantly less traumatic than the skin window method and more informative than the studies of venous blood.