Interconnections between Inflammageing and Immunosenescence During Ageing

Acute inflammation is a physiological response to injury or infection, with a cascade of steps that ultimately leads to recruitment of immune cells to clear invading pathogens and heal wounds. However, chronic inflammation arising from continued presence of the initial trigger, or dysfunction of signalling and/or effector pathways, is harmful to health. While successful ageing in older adults including centenarians is associated with low levels of inflammation, elevated inflammation increases the risk of poor health and death [1–3]. Hence inflammation has been described as one of seven pillars of ageing. Age-associated sterile, chronic, and low-grade inflammation is commonly termed inflammageing - it is not simply a consequence of increasing chronological age, but is also a marker of biological ageing, multimorbidity and mortality risk. While inflammageing was initially thought to be caused by “continuous antigenic load and stress”, reports from the last two decades describe a much more complex phenomenon also involving cellular senescence and ageing of the immune system. In this review, we explore the sources and consequences of inflammageing and highlight potential interventions. In particular, we assess the contribution of cellular senescence to age-associated inflammation, identify patterns of pro- and anti-inflammatory markers characteristic of inflammageing, describe alterations in the ageing immune system that lead to elevated inflammation, and finally assess the ways that diet, exercise and pharmacological interventions can reduce inflammageing and thus improve later life health.

Influence of feed factors on the dynamics of sow metabolism

The article presents the results of studies on the effect of increased fiber levels in combination with the probiotic Profort in SK-1 mixed feed on the formation of metabolism in the body of pregnant sows, in SK-2 mixed feed on the formation of microbiocenosis and nonspecific immunity in lactating sows in the conditions of the pig complex LLC "Agronika" of the Kursk region. The tested mixed feed SK-1 has an optimal structure of feed ingredients, which corresponds to the physiological state of deepfat sows. At the same time, the fiber content increases to 8.0%, the crude protein content is 176.3 g / kg at the exchange energy level of 10 MJ/kg. The SK-2 compound feed for lactating sows contains 12.87 MJ / kg, crude protein-18.3%, crude fat-4.86%, crude fiber-4.93%. The studies found that the blood serum of the experimental group of pregnant sows significantly (P < 0.05) higher glucose and total protein, the tendency of growth of gamma-globulins indicates an increase in the immunological reactivity of the sow body. There is a tendency to increase a-globulins and b-globulins. Indicators of enzyme activity in-dicate the intensity of metabolic processes in the body of deep-fat sows. The indi-cator of alkaline phosphatase, in the experimental group, tends to increase by 2.8 times, the decrease in the indicator of the amylase enzyme in the experimental group by 1.2 times. Increased lipase activity in the experimental group by 2.2 times. In studies of the facies environment of lactating sows, a positive trend of re-ducing the conditionally pathogenic microflora was established, while an increase in Bifidobacterium to 107 CFU/g was found, with a decrease in E. from 60 to 105 CFU / g, which indicates a positive dynamics of microbiocenosis due to an increase in normal microflora. Th ratio/Ts in the blood of sows of the experimental group increased due to an increase in the number of T-helpers by 73.8% , while reducing T-suppressors by 85.9 %. An increase in the blood content of animals of the second group of G-class immunoglobulins indicates a decrease in the antigenic load on the body.

Evaluation of antigenic load in pregnant sows and identification of isoimmunization markers in offspring to increase productive and reproductive indicators for obtaining biologically safe organic products

Immunological aspects of the relationship in the functional system “mother-fetus” are consciously important both in the scientific and practical direction of the life of living organisms. The formation of the main parameters of isoimmunization is an equivalent aspect in relation to the basics of the formation of artificial tolerance mechanisms. Changes in immunological reactivity to viral and bacterial antigens may cause increased susceptibility to infectious diseases. Different levels of this condition in newborn and adult animal organisms should be based on the fact that the fetus and newborn after birth first comes into contact with the antigen, while the adult body already has partial sensitization. Chronic carrier of pathogens in animals and their influence on the spread of the infectious process is an urgent problem of modern veterinary medicine. The possibility of vaccination in newborns is limited by the presence of maternal antibodies that have an immunosuppressive effect.The immunological relationship between the fetal mother’s body in a non-inbred population should be considered in two aspects: the effects caused by maternal antigens in the fetus; the effect of fetal antigens on the mother’s body. Level of functional reserves of the pregnant body is important in the prevention of intrauterine infection.

State of humoral immunity in lactating sows and suckling piglets

The state of humoral immunity in lactating sows and its formation in suckling piglets in the conditions of a pig-breeding complex were studied. In sows after farrow, a high content of total immunoglobulins, the main classes of IgG, IgM and IgA, the concentration of which decreased in a day, followed by an increase in serum as lactation continued was detected in the serum and colostrum. In piglets at birth, the content of total immunoglobulins and the main Ig isotypes was insignificant, and at the age of twenty-four hours, as a result of absorption of colostral immunoglobulins in the small intestine, their amount in the serum was the highest, followed by a decrease in the concentration of IgG and IgA until the end of the suckling period, and IgM - up to day 14 with a further increase in its content, indicating the formation of a primary immune response. It has been detected that the dominant class of immunoglobulins in the serum and colostrum of sows and the serum of piglets is IgG, and in milk - IgA. An increase in the level of medium-dispersed circulating immune complexes (C4%) and their relation to giant (C3%) CICs was revealed, associated with an increase of the antigenic load in sows in the second half of lactation as a result of immunization, in milk - with a decrease in the content of the main classes of immunoglobulins, and in piglets, due to this, with a decrease in passive immunity.

Enhancing mucosal immunity by transient microbiota depletion

Tissue resident memory CD8+ T cells (Trm) are poised for immediate reactivation at sites of pathogen entry and provide optimal protection of mucosal surfaces. The intestinal tract represents a portal of entry for many infectious agents; however, to date specific strategies to enhance Trm responses at this site are lacking. Here, we present TMDI (Transient Microbiota Depletion-boosted Immunization), an approach that leverages antibiotic treatment to temporarily restrain microbiota-mediated colonization resistance, and favor intestinal expansion to high densities of an orally-delivered Listeria monocytogenes strain carrying an antigen of choice. By augmenting the local chemotactic gradient as well as the antigenic load, this procedure generates a highly expanded pool of functional, antigen-specific intestinal Trm, ultimately enhancing protection against infectious re-challenge in mice. We propose that TMDI is a useful model to dissect the requirements for optimal Trm responses in the intestine, and also a potential platform to devise novel mucosal vaccination approaches.

A Candidate Multi-Epitope Vaccine Against Pathogenic Chandipura Vesiculovirus Identified using Immunoinformatics.

Chandipura vesiculovirus (CHPV) is a rapidly emerging pathogen responsible for causing acute encephalitis. Due to its widespread occurrence in Asian and African countries, this has become a global threat, and there is an urgent need to design an effective and non-allergenic vaccine against this pathogen. The conventional method of vaccine design involves large proteins or whole organism which leads to unnecessary antigenic load with increased chances of allergenic reactions. In addition, the process is also very time consuming and labour intensive. These limitations can be overcome by peptide-based vaccines comprising of short immunogenic peptide fragments that can elicit highly targeted immune responses, avoiding the chances of allergenic reactions, in a relatively shorter time span. The multi-epitope vaccine constructed using CTL, HTL and IFN-γ epitopes was able to elicit specific immune responses when exposed to the pathogen, in-silico. Not only that, Molecular Docking and Molecular Dynamics Simulation studies confirmed a stable interaction of the vaccine with the immune receptors. Several physicochemical analyses of the designed vaccine candidate confirmed it to be highly immunogenic and non-allergic. The computer-aided analysis performed in this study suggests that the designed multi-epitope vaccine can elicit specific immune responses and can be a potential candidate against CHPV.

Polymorbidity, ageing of immune system and low-grade systemic inflammation: a challenge for modern medicine

The review article considers the data from literature that concern polymorbidity aspects, its interrelations with ageing of immune system and lo-grade immune ageing, mechanisms of genesis, approaches to its prevention and treatment. Evolution of “comorbidity” and “polymorbidity” terms is traced, an updated definition of polymorbidity is proposed. The world-wide incidence of polymorbidity is increased and now it reaches 23-25% in general population, and up to 98%, in elderly people (> 65 years old). The risk factors of polymorbidity are considered, like as its social burden due to high costs for healthcare, high mortality rates, excessive treatment provided by multidisciplinary specialists. We present evidence for common molecular and cellular mechanisms involved in ageing and polymorbidity, being unified by the term “inflammaging” which represents a low-grade chronic systemic inflammation associated with ageing. The data are presented that concern the “inflammaging” development with involvement of ageing cells from innate and adaptive immunity systems, different pro and anti-inflammatory mediators, lifelong antigenic load. The data are analyzed concerning functional and structural changes in the inborn and adaptive immune system in ageing, role of these changes in “inflammaging” persistence and development of polymorbid conditions. There are complex interactions shown between the bodily senescence and immune ageing, with similar underlying mechanisms in some cases, however, being quite different in other instances. With age, upon existing risk factors, the changed adaptive immunity in most people is not able to full-scale coping with chronic antigenic load, thus increasing the risk of diseases. Moreover, in many elderly people these changes are compensated by steady activation of the innate immunity cells. It is noted that the aging events and development of disease (polymorbidity) cannot be considered distinct entities, since they can interact, being, however, basically different in their nature. In future, one should concentrate our efforts on elucidation of molecular and cellular mechanisms of these interactions, solution of the tasks oriented for development of such interventions that could be able to reduce harmful consequences of ageing and to use useful effects for health maintenance and reaching maximal longevity.

Gut Microbial-Derived Metabolomics of Asthma

In this review, we discuss gut microbial-derived metabolites involved with the origins and pathophysiology of asthma, a chronic respiratory disease that is influenced by the microbiome. Although both gut and airway microbiomes may be important in asthma development, we focus here on the gut microbiome and metabolomic pathways involved in immune system ontogeny. Metabolite classes with existing evidence that microbial-derived products influence asthma risk include short chain fatty acids, polyunsaturated fatty acids and bile acids. While tryptophan metabolites and sphingolipids have known associations with asthma, additional research is needed to clarify the extent to which the microbiome contributes to the effects of these metabolites on asthma. These metabolite classes can influence immune function in one of two ways: (i) promoting growth or maturity of certain immune cell populations or (ii) influencing antigenic load by enhancing the number or species of specific bacteria. A more comprehensive understanding of how gut microbes and metabolites interact to modify asthma risk and morbidity will pave the way for targeted diagnostics and treatments.

Detailed Multi-Method Analysis of Bone Marrow from Pediatric Pre-B-ALL Patients Prior to CD19-CAR-T Therapy Subsequently Evidencing Overt CAR-T Resistance

Little is known about the disease-specific factors that predict responsiveness to CAR-T cell therapy, other the direct presence of the CAR-T target. Clinical outcomes at our center have demonstrated that durable responses to CD19-directed CAR-T therapy in pediatric pre-B-ALL (acute lymphoblastic leukemia) are associated with persistence of CAR-T cells in the peripheral blood, antigenic load (percent CD19-positive cells in marrow prior to CAR-T infusion), and apheresis product T-cell quality (Finney O, et al., 2019). However, in a small number of cases where both antigenic load and T-cell quality predicted a good response, the treatment failed rapidly. This led us to undertake a detailed investigation of the leukemia itself in order to discover potential disease-associated factors that correlate with resistance to CAR-T therapy. We employed advanced exomic, and single-cell genomic and epigenomic analysis techniques to define signatures present in four CD19-CAR-T resistant bone marrow biopsy specimens, in comparison to five specimens from CD19-CAR-T responsive disease, from patients enrolled in a phase I clinical trial at Seattle Children's Hospital (PLAT-02, NCT02028455). Current cytogenic approaches to identify high risks markers (Ph+, Ph-like, MLL) were not informative as to CAR-T susceptibility, as high risk leukemias were CAR-T responsive; while a CAR-T resistant leukemia contained a marker (ETV6-RUNX1 fusion) previously associated with a good prognosis. Thus, we performed bulk whole-exome sequencing and RNAseq, single cell (sc) RNAseq, sc B-cell receptor (BCR)-seq, methylation array, H3K27ac ChIPseq, and ATACseq on these marrow samples. Initial genomic analysis revealed a total of 5 previously described hotspot mutations in ABL1, IKZF1, EP300, and 2 in KRAS. RNAseq analyses identified actionable fusions for ABL1, ETV6, ETV5, and KMT2A. Interestingly, a therapy-sensitive leukemia harbored a KMT2A-AFF1 fusion that was shown to predispose patients to leukemic plasticity and lineage switching when treated with blinatumomab. Importantly, we identified CREBBP-fusions in leukemias that failed to achieve CD19-CAR-T cell induced B cell aplasia. CREBBP perturbations have previously been associated with relapsed and refractory ALL, but not with resistance to CAR-T therapy. Single cell RNAseq and scBCRseq data are being analyzed for the existence of mixed lineage and gene expression-based heterogeneity that may predict clonal selection under CAR-T pressure. RNASeq analysis identified upregulation of JUN and JUND transcripts in CAR-T resistant disease, a finding which is complemented by the hypermethylation of JUND in CAR-T sensitive disease. Similarly, ATACseq and methylation data is being analyzed for lineage specification in CAR-T resistant leukemia. In comparing dysfunctional to functional CAR-T responders by ATACseq, > 10,000 unique open chromatin regions were identified in dysfunctional responders, as opposed to <500 open chromatin regions in the functional responders, indicating that CAR-T resistant disease had more open chromatin. A recent published analysis of cancer cell lines identified EP300 and CREBBP mutations that were proposed to increase substrate acetylation, and which "…may represent the first cancer-associated gain of function mutations for p300 and CBP…" (Ghandi M, et al, 2019). Our study represents one of the most comprehensive approaches to genomic profiling for B-ALL patient samples to date. The immune evasion we have described is not due to overt CD19 antigen loss, or to a long-term process of genetic alteration or drift. We propose that continued analysis of our data may reveal that epigenetic plasticity is a component of CAR-T resistance. The presence of CREBBP fusion genes or mutations, methylation array-based identification of altered JUND/JUN regulation, and ATAC Seq identification of multiple open regions of the genome in leukemias from dysfunctional responders lend support to this hypothesis. Although our analysis is preliminary and the sample number is small, we believe these in-depth analyses will highlight crucial differences in leukemia that predict responsiveness to CAR T therapy Disclosures Gardner: Novartis: Honoraria. Jensen:Bluebird Bio: Research Funding; Juno Therapeutics, a Celgene Company: Research Funding. Orentas:Lentigen Technology Inc., a Miltenyi Biotec Company: Consultancy, Research Funding.