Duodenal Eosinophilia in Functional Dyspepsia

Aim: to present observation of a patient diagnosed with functional dyspepsia based on current guidelines, and having increased eosinophil counts in the biopsy specimen of duodenal mucosa. To consider possible causes of duodenal eosinophilia in the light of present-day concepts.Highlights. Patient K., 40 years old, complained of dyspeptic phenomena, the first appearance of which she had noted at the age of 18. The patient noted poor tolerance to canned and fermented foods, which provoked an increase in dyspepsia and sometimes caused watery diarrhea. The examination excluded “symptoms of concern”. Successful antihelicobacter eradication therapy was carried out. Morphological examination of the stomach showed phenomena of mild chronic inflammation without intestinal metaplasia or glandular atrophy. A biopsy of the mucosa of the descending part of the duodenum showed a moderate increase in the levels of mononuclears and eosinophils in its lamina propria without penetration into the epithelium of the villi or formation of clusters. The patient suffers from pollinosis; sensitization to birch pollen was diagnosed by a skin prick test. However, she has no oral allergy symptoms, which does not allow linking duodenal eosinophilia to food allergy. Based on current guidelines, the patient was diagnosed with functional dyspepsia. In addition to dietary restrictions, treatment courses with a proton pump inhibitor, itopride, and S-methylmethionine sulfonium chloride, which has an antihistamine effect, were recommended for periods of worsening dyspepsia.Conclusion. The clinical significance of duodenal eosinophilia and local histamine production in patients with a clinical diagnosis of functional dyspepsia deserves special attention. Triggering factors provoking the worsening of symptoms should be analyzed; in particular, a food diary and exclusion of food allergies are recommended. Histamine-neutralizing drugs may play a role in the treatment of FD with duodenal eosinophilia in the future

Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs

PolG-alpha is a nuclear-encoded enzyme which provides replication and repair of mitochondrial DNA. D257A mutation of PolG-alpha leads to change in the N-terminal "proofreading" domain, which deprives the enzyme of 3′-5′ exonuclease activity, resulting in accumulation of mutations in the mitochondrial genom

On the way from SARS-CoV-sensitive mice to murine COVID-19 model

The coronavirus disease 2019 (COVID-19) is a master killer which appeared suddenly and which has already claimed more than 200,000 human lives. In this situation, laboratories are in urgent need for a COVID-19 murine model to search for effective antiviral compounds. Here we propose a novel strategy for the development of mice that can be inoculated by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the COVID-19 causative agen

ФУНКЦИОНАЛЬНЫЙ ПОТЕНЦИАЛ КЛЕТОК ПАМЯТИ CD8+ В УСЛОВИЯХ ЛИМФОПЕНИИ, ВЫЗВАННОЙ ВВЕДЕНИЕМ ГИДРОКОРТИЗОНА

Background. Wide use of glucocorticoids therapy for neoplasms, autoimmune diseases and allergies is associated with suppression of adaptive immunity that requires profound study of their immunoregulatory properties and immunotoxicity.Results. In this work, using our model of selective activation of mouse CD8+ memory cells in the mixed lymphocyte reaction (MLR) in vitro, we show for the first time that intraperitoneal injection of high dose hydrocortisone (2.5 mg per animal) allows to detect memory cells in the thymus of animals immunized with allogeneic tumor cells. Similar to memory cells from other lymphoid organs, hydrocortisone-resistant thymic lymphocytes from immune animals respond on allogeneic stimulators subjected to severe heat shock and are immunologically specific to immunizing alloantigen. Thus, cortisone-resistant thymocytes are partially or completely represented by memory cells. We also show here that memory responses of heterozygotes on TCR a-chain knock-out (genetically incapable to secondary rearrangement of TCR achains) are significantly enhanced as compared with the ones of wild type mice.Conclusion. These findings allows to suggest the hypothesis according to which memory T cell clones proliferating in primary immune response migrate into thymus providing necessary microenvironment for reexpression of recombinases. After editing of genes encoding TCR achains, such T lymphocytes can return to peripheral repertoire maintaining its wideness.Введение. Широкое применение глюкокортикоидов в терапии онкологических, аутоиммунных и аллергических заболеваний сопряжено с подавлением функций адаптивного иммунитета и ставит задачу углубленного исследования их иммунорегуляторных свойств и иммунотоксичности.Результаты. Используя разработанную нами модель селективной активации мышиных клеток памяти CD8+ в смешанной культуре лимфоцитов (mixedlymphocytereaction, MLR) invitro, в этой работе мы впервые показали, что внутрибрюшинное введение мышам гидрокортизона в высокой дозе (2,5 мг на 1 животное) позволяет обнаружить Т-клетки памяти в тимусе животных, иммунизированных клетками  аллогенной опухоли. Как и клетки  памяти из других лимфоидных органов, лимфоциты тимуса иммунных животных, резистентные к гидрокортизону, отвечают пролиферацией на аллогенные стимуляторы, подвергнутые острому тепловому шоку и иммунологически специфичны к иммунизирующему аллоантигену. Таким образом, кортизонрезистентные тимоциты частично или полностью представлены клетками памяти. Также мы показали, что у гетерозигот по нокауту α-цепи TCR (генетически неспособных ко вторичной реаранжировке α-цепей) ответы клеток памяти значительно усилены по сравнению с ответами клеток памяти мышей дикого типа.Заключение. Полученные данные позволяют выдвинуть гипотезу, согласно которой клоны Т-клеток памяти, размножившиеся в первичном иммунном ответе, мигрируют в тимус, обеспечивающий микроокружение, необходимое для реэкспрессии ими рекомбиназ. После завершения редактирования α-цепей TCR такие Т-лимфоциты могут вернуться в периферический репертуар, поддерживая его широту

“Usage a CRISPR/Cas9-based for obtaining knockouts of economically significant cattle genes”

An experimental work dealing with the gene modification using the Cas9 RNA-based editing system was performed. Point site-specific breakpoints in gDNA were introduced at the zygote stage by microinjection of spCas9 mRNA protein and guide RNAs into the zygote cytoplasm. Oocytes that extruded the first and second polar bodies were used for the injection. 2 series of microinjections of gene editing designs for early bovine embryos were made. The degeneration ranged from 10% to 56% in different groups. A total of 100 injections were performed. Cleavage was started by 78% of the surviving oocytes; 5 embryos reached the blastocyst stage, which was 16% of the number of dividing embryos. All the resulting embryos were analyzed to evaluate the efficiency of editing. gDNA was isolated from all embryos that had reached the blastocyst stage. Using Sanger sequencing of genes of interest in pre-implantation bovine embryos and biopsies from them, it was shown that in 5 out of 17 embryos resulting from microinjections of guide RNA against the BLG gene and SpCas9 mRNA, and in 2 out of 9 embryos after microinjections of guide RNA against CD209 gene and SpCas9 mRNA, the required genome modifications were found. This is indicative of the high efficiency of this delivery method of the editing system

Using CRISPR/Cas9 for generation the cd209 knockout is a way to get cattle breeds resistant to the Bovine leukemia virus (BLV)

Bovine leukemia virus (BLV) causes enzootic leukemia - a chronic infectious disease occurring against the background of embedding the virus in the genome of B-lymphocytes and leading to malignization, invasion of tumor cells in organs and the formation of tumors. The disease is common in the United States, Japan, and Asia. In Russia, up to 30% is infected with BLV. Moreover, there is evidence of the presence of antibodies to the BLV virus in some groups of people, and the relationship between BLV and cancer in humans is widely discussed. All this indicates an urgent need to study BLV and create breeds resistant to it. The development of approaches to solving this problem is complicated by the fact that the receptor through which the infection is carried out is still unknown. Recently, it has been suggested that the virus penetrates the animal's lymphocytes using the CD209 molecule. In this paper, we propose a genome editing system based on CRISPR/Cas9 to get a knockout for this gene. We assume that animals obtained using the presented genome editing system will be resistant to infection with the bovine leukemia virus

Metabolic aspects of adoptive immunotherapy of tumors

In wide number of approaches to treatment of cancer immunotherapy plays special role. This approach exploits capabilities of immune system to support genetic constancy of different cells and tissues of the organism. Immunotherapy is designed to induce tumor cell destruction  by T-lymphocytes whose receptors can recognize peptides of mutant proteins complexed with the molecules of the major histocompatibility complex. In clinical practice T-lymphocytes can result in sustained and complete responses in patients whose cancers were resistant to available treatment options. Recent evidences suggest that efficiency of such therapy generally depends on metabolic properties of T-lymphocytes. A number of approaches allows modulate T-cell metabolism providing strategies to optimize activity of anti-tumor T-lymphocytes

Gene editing CRISPR/Cas9 system for producing cows with hypoallergenic milk on the background of a beta-lactoglobulin gene knockout

Beta-lactoglobulin is the main allergen of cow’s milk. Modern approaches to reducing the allergenicity of milk require significant costs for its fermentation. An alternative approach could be the creation of productive breeds with a knockout for the gene of this protein. This will allow, at no additional cost, to increase its safety for children who are breast-fed. In this article, we report on the development of a system for gene editing to create cows with a knockout of the beta-lactoglobulin gene

FUNCTIONAL CAPACITY OF MEMORY CELLS CD8+ UNDER LYMPHOPENIA INDUCED BY INJECTION OF HYDROCORTISONE

Background. Wide use of glucocorticoids therapy for neoplasms, autoimmune diseases and allergies is associated with suppression of adaptive immunity that requires profound study of their immunoregulatory properties and immunotoxicity.Results. In this work, using our model of selective activation of mouse CD8+ memory cells in the mixed lymphocyte reaction (MLR) in vitro, we show for the first time that intraperitoneal injection of high dose hydrocortisone (2.5 mg per animal) allows to detect memory cells in the thymus of animals immunized with allogeneic tumor cells. Similar to memory cells from other lymphoid organs, hydrocortisone-resistant thymic lymphocytes from immune animals respond on allogeneic stimulators subjected to severe heat shock and are immunologically specific to immunizing alloantigen. Thus, cortisone-resistant thymocytes are partially or completely represented by memory cells. We also show here that memory responses of heterozygotes on TCR a-chain knock-out (genetically incapable to secondary rearrangement of TCR achains) are significantly enhanced as compared with the ones of wild type mice.Conclusion. These findings allows to suggest the hypothesis according to which memory T cell clones proliferating in primary immune response migrate into thymus providing necessary microenvironment for reexpression of recombinases. After editing of genes encoding TCR achains, such T lymphocytes can return to peripheral repertoire maintaining its wideness