Identification of inflammatory markers in eosinophilic cells of the immune system: fluorescence, Raman and CARS imaging can recognize markers but differently

Eosinophils (Eos) play an important role in the immune system’s response releasing several inflammatory factors and contributing to allergic rhinitis, asthma, or atopic dermatitis. Since Eos have a relatively short lifetime after isolation from blood, usually eosinophilic cell line (EoL-1) is used to study mechanisms of their activation and to test therapies. In particular, EoL-1 cells are examined in terms of signalling pathways of the inflammatory response manifested by the presence of lipid bodies (LBs). Here we examined the differences in response to inflammation modelled by various factors, between isolated human eosinophils and EoL-1 cells, as manifested in the number and chemical composition of LBs. The analysis was performed using fluorescence, Raman, and coherent anti-Stokes Raman scattering (CARS) microscopy, which recognised the inflammatory process in the cells, but it is manifested slightly differently depending on the method used. We showed that unstimulated EoL-1 cells, compared to isolated eosinophils, contained more LBs, displayed different nucleus morphology and did not have eosinophilic peroxidase (EPO). In EoL-1 cells stimulated with various proinflammatory agents, including butyric acid (BA), liposaccharide (LPS), or cytokines (IL-1β, TNF-α), an increased production of LBs with a various degree of lipid unsaturation was observed in spontaneous Raman spectra. Furthermore, stimulation of EoL-1 cells resulted in alterations of the LBs morphology. In conclusion, a level of lipid unsaturation and eosinophilic peroxidase as well as LBs distribution among cell population mainly accounted for the biochemistry of eosinophils upon inflammation.

Eosinophilic endometrial metaplasia - case report and brief literature review on its immunohistochemical characteristics

Endometrial epithelial metaplasia is described as transition of the normal endometrial epithelial cells by benign complex proliferation of cells. These metaplastic changes have been frequently reported as associated changes in endometrial hyperplasia and adenocarcinoma more than non-neoplastic samples and are also known to appear atypical occasionally, and hence can be a diagnostic challenge. Eosinophilic cell change is one of the most frequently encountered endometrial metaplasias. Eosinophilic syncytial change is a form of eosinophilic endometrial metaplasia, and is known to mimic endometrial serous carcinoma, again posing a diagnostic challenge. In this article, we have presented a case of endometrial eosinophilic metaplasia in a 47-year-old patient along with a brief discussion on immunohistochemical characteristics of eosinophilic syncytial change that could help pathologists to differentiate them from malignancies in challenging scenarios.

Relevance of tissue eosinophilia in subdural hematomata

Chronic subdural hematomata (CSDH) are treated by evacuation. Recurrence occurs in 3-20% of cases, but the factors determining its occurrence have not been determined. Having observed that eosinophil cell infiltrates are often present in the outer membrane of CSDH, our aim was to determine whether such infiltrates are associated with risk of recurrence. Histological sections of the resections from 72 patients with primary CSDH (Mean age 73.4) and 16 with recurrent CSDH (Mean age 72.1) stained with H&E were graded by blinded observers for eosinophilic cell infiltrates using a semiquantitative 0 to 3 scale. The risk of recurrence requiring reoperation (RrR) in primary CSDH was 11.1%, and 12.5% in recurrent CSDH (meaning third surgery was required). A dense (grades 2 or 3) eosinophilic infiltrate was present in 22.2% of primary CSDH; the RrR was 0% in these cases, as compared with 14.8% in cases with sparse (grades 0-1) eosinophilic infiltrate. Among recurrent CSDH cases, 12.5% (2/15) showed a dense eosinophilic infiltrate; the RrR was also 0%, contrasting with 14.3% in those with sparse eosinophilic infiltrate. We conclude that eosinophils either play a role or are a marker of a process leading to stabilizing CSDH, making them less prone to rebleeding. Abstract not previously publishedLearning ObjectivesDescribe the risk of recurrence following surgical evacuation of chronic subdural hematomataRecognize the variable presence of eosinophils in chronic subdural hematomataCite the presence of eosinophils is predictive of absence of recurrence

Model of hypereosinophilic syndrome in superinvasive opistorchosis. Report I. Liver pathology

Hypereosinophilic syndrome (HES) was modeled in mature Syrian hamsters of both sexes by the means of infecting them with O. felineus metacercariae (100 larvae); there were two superinvasions, of 100 larvae each. The animals were butchered on 10th day after invasion, the first superinvasion and, on 24th day after the second superinvasion. The material (liver) was stained by histological methods, the IHC method was used. The manifestations of HES were revealed: extensive fields of eosinophilic cell proliferates (index – 0.52), the absence of proliferation and differentiation of progenitor cells into elements of cholangio- and hepatocellular diferons (CCD and HCD). After the second superinvasion, a depletion of the liver’s replicative potential was noted: in the areas of infiltrates from eosinophils, the proliferation of connective tissue, diffuse and focal fibrosis were observed.

Nonclinical Safety Assessment of an Inhaled Formulation of Serelaxin: A Recombinant Human Protein in Rats and Cynomolgus Monkeys (Macaca fascicularis)

Serelaxin is a recombinant human relaxin-2 intended for cardiovascular indications. Inhalation was chosen as alternative route to intravenous to allow daily administration for chronic applications and home treatment. A total of 4 short-term studies were conducted in rats and cynomolgus monkeys with inhaled formulation of serelaxin at dose up to 10 mg/kg/d. All rats and cynomolgus macaques receiving serelaxin were exposed to the test item. One rat and approximately 50% of macaques developed immunogenicity, which did not appear to affect exposure. No adverse effect on respiratory function or systemic changes was noted. Both species developed similar microscopic lesions characterized by eosinophilic cell infiltration around bronchi; however, in the rat, this was more pronounced and extended to a perivascular location. In addition, in the rat, serelaxin showed eosinophilic crystalline material associated with macrophages in the alveoli and bronchioles. In macaques, serelaxin induced minimal macrophage infiltrates in alveoli and perivascular/peribronchiolar mononuclear cell infiltrations. The minimal airway eosinophilic/mononuclear inflammatory cell infiltrations were considered to be nonadverse in macaques due to the minimal severity and the lack of any other alterations in the lung parenchyma. In the rat, the presence of eosinophilic crystalline material and macrophage response, characterized as precipitated test article, was considered adverse.

Appendiceal neuroma: an uncommon entity

Appendiceal neuroma or neurogenous hyperplasia of the appendix is an uncommon entity. We report an incidental finding of an appendiceal neuroma in a 35 year male patient who underwent appendectomy. On microscopy showed thickened appendicular wall with well circumscribed submucosal nodules of proliferating spindle cells in myxoid areas, eosinophilic cell infiltration and obliterative appendicitis. It was reported as appendiceal neuroma. We are presenting this case for its uncommon entity, clinical and histopathological finding.

Winter Exercise Reduces Allergic Airway Inflammation: A Randomized Controlled Study

Background: Physical exercise is often recommended as additional treatment for people suffering from allergic rhinitis and/or asthma, but less is known about the specific effects of recreational winter outdoor exercise on allergic airway inflammation. Methods: We performed a longitudinal, randomized controlled intervention study to investigate the effects of recreational winter exercise on allergic airway inflammation, quality of life, spirometry and cardiorespiratory fitness in adults suffering from allergic rhinitis and/or asthma. The exercise group participated in a ten-day winter sports program. The control group did not receive any intervention. Results: A significant improvement of fractional oral exhaled nitric oxide (FeNO; p = 0.008, day 10) and a significant decrease in FeNO after a single 4 h hiking tour (p < 0.001, time effect) were observed for the exercise group. The nasal eosinophilic cell count revealed a short-term reduction (p = 0.021, treatment effect) in the exercise group and for the visual analogue scale sustainable improvements in allergic symptoms (p < 0.001, day 60) were found. No adverse effects of outdoor winter exercise were observed. Conclusion: Recreational winter exercise at moderately cold temperatures reduces allergic airway inflammation measured as FeNO, nasal eosinophilic cell count and induces sustainable improvements in allergic symptoms.

Hypereosinophilic syndromes

Background: The last few years have seen a complete change in the etiopathogenetic features, classification and therapeutic approach of the hypereosinophilic syndrome (HES), a multiorgan targeted blood disease. The discovery of a genetic mutation and the occurrence of a new fusion gene, named FIP1L1-PDGFRA (FIP gene), in some patients allowed the identification of a new myeloproliferative disorder, M-HES: thereafter, the pivotal therapeutic role of the tyrosine kinase inhibitors, particularly, imatinib mesylate, was clearly detected. In the same period a new pathogenetic mechanism has been detected: some authors described the presence of a CD3-CD4 +Tcell clone correlating with the overproduction of IL5, a potent eosinophilic cell line stimulating cytokine. As a consequence an international consensus committee proposed a new classification for these syndromes, in accordance with these new pathogenetic features. The disease is characterized by an extensive tissue and organ damage due to an eosinophilic cell infiltration and leading to the release of toxic cytokines and subsequent organ dysfunction. The heart, lungs, gastrointestinal apparatus, skin and central nervous system are affected. Moreover the released cytokines can induce a thrombophilic status and thromboembolic events can occur throughout the body. Aim of the study: We describe the diagnostic procedures that are necessary in order to obtain a correct diagnosis and classification of the disease and to evaluate the presence of an organ and tissue damage. In particular, bone marrow biopsy and cytogenetic examination of blood and marrow are necessary for detecting M-HES cases that are positive for the FIP gene. In these patients, imatinib mesylate has a leading role for obtaining complete remission of the disease in a high percentage of cases. We also examine the therapeutic options for the other forms of the disease: prednisone, interferon, hydroxiurea are effective therapeutic tools in these patients. Finally, the new therapeutic perspectives, such as monoclonal antibodies directed against IL 5 or IL 5 receptor and novel tyrosine kinase inhibiting drugs, are examined.