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TCR-T cell therapy represents a promising advancement in adoptive immunotherapy for cancer treatment. Despite its potential, the development and preclinical testing of TCR-T cells face significant challenges. This review provides a structured overview of the key stages in preclinical testing, including in silico, in vitro, and in vivo methods, within the context of the sequential development of novel therapies. This review aimed to systematically outline the processes for evaluating TCR-T cells at each stage: from in silico approaches used to predict target antigens, assess cross-reactivity, and minimize off-target effects, to in vitro assays designed to measure cell functionality, cytotoxicity, and activation. Additionally, the review discusses the limitations of in vivo testing in animal models, particularly in accurately reflecting the human tumor microenvironment and immune responses. Performed analysis emphasizes the importance of these preclinical stages in the safe and effective development of TCR-T cell therapies. While current models provide valuable insights, we identify critical gaps, particularly in in vivo biodistribution and toxicity assessments, and propose the need for enhanced standardization and the development of more representative models. This structured approach aims to improve the predictability and safety of TCR-T cell therapy as it advances towards clinical application., (© 2024. The Author(s).)

The transcriptomic profile associated with osteo- and adipogenic differentiation in growth-arrested multipotent mesenchymal stromal cells (MSCs) from human adipose tissue was analyzed in vitro at 20% (standard laboratory) and 5% (tissue-related) O 2 levels. Compared with day 7, at 5% O 2 on day 14 spontaneous upregulation of osteo- (RUNX2, SP7, BGLAP, and SPP1) and adipogenic differentiation (CEBPA, PPARG, and ADIPOQ) genes in MSCs was observed (p < 0.05). Thus, upon expansion under tissue-related O 2 , MSCs demonstrated a bipotent transcriptomic profile, which may contribute to the improvement of their hematopoiesis-supportive function.

Objectives: The aim of the study was to evaluate the potential contribution made by tumor necrosis factor (TNF) autoantibodies to the pathogenesis of bronchial asthma (BA)., Methods: We used affinity chromatography methods and a magnetic separation procedure to purify human autoantibodies specific to TNF. The autoantibodies were used as a calibration material to determine the absolute content of autoantibodies to TNF using enzyme-linked immunosorbent assay (ELISA). TNF content and levels of soluble receptors to TNF were determined using the ELISA commercial test kits., Results: We demonstrated significant increases in the levels of TNF and soluble TNF receptors in the sera of patients with uncontrolled and controlled BA, as compared with healthy donors. Levels of autoantibodies of the IgG2 and IgG4 subclasses were significantly higher in sera from patients with uncontrolled BA than in healthy donors. Levels of IgG2 autoantibodies were significantly higher in sera from patients with uncontrolled BA than in patients with controlled BA., Conclusions: BA is associated with changes in the levels of not only TNF and soluble receptors for TNF, but also autoantibodies to TNF. Given the magnitude of the changes in the levels of different subclasses of autoantibodies to TNF, we propose that these autoantibodies might contribute to the pathogenesis of BA.

Autoantibodies to cytokines are important biological effector molecules that can regulate cytokine activities. The aim of the study was to develop a protocol to purify autoantibodies to tumor necrosis factor from human serum, for use as a calibration material to determine the absolute content of autoantibodies to tumor necrosis factor by enzyme-linked immunosorbent assay. The proposed protocol includes a set of affinity chromatography methods, namely, Bio-Gel P6DG sorbent to remove albumin from serum, Protein G Sepharose 4 Fast Flow to obtain a total immunoglobulin G fraction of serum immunoglobulins, and Affi-Gel 15 to obtain specifically antibodies to tumor necrosis factor. The addition of a magnetic separation procedure to the protocol eliminated contaminant tumor necrosis factor from the fraction of autoantibodies to tumor necrosis factor. The protocol generated a pure fraction of autoantibodies to tumor necrosis factor, and enabled us to determine the absolute concentrations of different subclasses of immunoglobulin G autoantibodies to tumor necrosis factor in apparently healthy donors., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Component communities of parasites of Phoxinus phoxinus from the Chovju River near Syktyvkar City (Komi Republic) are investigated in seasonal aspect. Material for the study has been collected in May-September 2000 from 90 host specimens of ages 0+-1+. Calculations of diversity indices and other coefficients have been made by two ways -- for metazoan parasites only and for whole parasite community including protozoan species. In both these cases three states of the community have been established, 1) forming community in May-June, 2) completely formed community in the end of June-July, 3) disintegrating community in August, 4) forming community in September. Seasonal dynamics of the component communities in young minnow thus found to be different from that in adult fish hosts.

A comparative analysis of the structure of component parasite communities in the minnow from ecologically safe or polluted to variable extent reservoirs was made. The analysis showed that the increase of the reservoir pollution by everyday drains, washings of fertilizers from the fields around and the dacha sections causes the change of dominant parasite species in the minnow, alteration of the sum of errors in the regression equation characterizing the spread of values of biomasses of the species forming the parasite community, and change of the "graphic" structure of the community. It is proved, that the high concentration of biogens has a destructive effect on natural systems, and at the same time causes the process of self-organizing, leading to the alteration of the community structure. As soon as even a small part of biogens is stopped to come into the reservoir, and the quality of environment is restored, the community restores its structure.

The material represented by 75 specimens of minnow of the age 2-2(+) was collected according to the standard technique in the Chovju River (tributary of the Vychegda River, a region of the settlement Nizniy Chov, district of Syktyvkar town) during the period June-September 2000. The quantitative estimation of the structure of the component parasite communities was performed by calculating errors of the equation of regression for each species group separately, with subsequent summarizing of means of errors by all parasite groups comprising the community (Dorovskikh, 2001 6; 2002 B). In order to have a possibility to get data complementing each other, the calculation of variety indexes and other indexes have been made for metazoan parasites only and for the whole community, including the protozoan parasites. In both cases, i.e. considering only metazoan parasites and the whole composition of parasite species, three states of the component parasite community have been recognised: the formed community (June), the community in destroying (July and August), the community in the process of formation (September). In the course of working on the total species composition we recorded the beginning of community destroying on 30th of June, and the beginning of community formation in August. Considering only the metazoan parasites, the community is defined (after: Pugachev, 1999) as the mature (balanced) one in June, while in July, August and September, it was unmature (off-balance) by its characteristics. Regarding the protozoan parasites, the community in June, August and September was characterized as the mature one based on indices of parasite biomass, and as unripe one based on the number of parasite individuals; however in July, both groups of indices allowed to refer it to the unripe state. However, these unripe states are essentially different. In July, it is the result of dieing out the parasites of the past generation; in August and September, it is the result of the appearances of new generations. Therefore we recognise three states of parasite community named above. Considering the whole species composition of parasites we noted the greater difference of index values based on the parasite specimen numbers and their conventional biomass, that was in the case of the metazoan parasites only. It is particularly noticeable in the middle of the June, in the period of the formed community. This fact, together with high errors of the equations of regression and the presence of the high number of Apiosoma, points to the disturbance in the structure of component parasite community in the minnow from the Chovju River. This is easily explicable, because the Chovju River is the polluted reservoir, and pollution comes from agricultural fields, Verhny-Chov settlement, pigsties and cow-sheds. The pollution is a seasonal factor here. The most powerful pollution was noted at May-June, and then it decreased along the beginning of rains in the end of August; in September it increased again. It is important to point out that the monitoring of the metazoan parasites only allows to reveal the general dynamics of the community during the period of observation, but does not allow to catch the beginning of its destroying and developing and to notice possible disturbances in the community structure caused by pollution of reservoirs, particularly, if this pollution is a seasonal factor as in the Chovju River. Three named states of the component community of the fish parasites take place in other periods of year than this observed in the intestional parasite communities of fish helminths of the temperate climate zone. The developing of parasite communities of the intestinal helminths of the Anguilla anguilla in England (Kennedy, 1997) and of Leuciscus idus from the Rybinsk reservoir (Zhohov, 2003) starts in the beginning of summer. In May, their species diversity is minimal and in August is maximal. In conditions of the middle stream of the Vychegda River, the species diversity of parasite community associated with the minnow is maximal in June and minimal in August, when it only begins developing.

The activity of natural killers and the capacity of peripheral blood mononuclears for the extracellular killing of Candida cells in patients with different forms of candidiasis has been determined in the radiometric test with the use of targets labeled with 3H-uridine RNAase and in the fungicidal test. On the basis of the data thus obtained the conclusion has been made that in candidiasis changes in the activity of natural killers and fungicidal capacity depend on the severity of the disease and the stage of the process. Linear correlation between the activity of natural killers and fungicidal capacity in candidiasis patients at the stage of exacerbation and in healthy persons is absent.

To compare the clinical picture and the immunological characteristics, 58 candidiasis patients differing by the severity and dissemination of the disease were examined. Chronic candidiasis of the skin and mucous membranes, the most severe and disseminated form of the disease, is associated with a decrease in the number of T-lymphocytes and changes in their subpopulations, as well as high titers of Candida albicans antigen and antibodies to it in blood sera. The immune system of patients with visceral candidiasis and chronic vulvovaginal candidiasis was similar to that of healthy persons in the characteristics under study. Immediate and mixed hypersensitivity occurred in candidiasis patients more frequently than in healthy persons. In extremely severe forms of Candida infection immediate hypersensitivity prevailed.

Two methods for measuring the activity of antibody-dependent cell-mediated cytotoxicity are compared. One of them is based on the determination of the relative amount of hemoglobin released from lysed sheep target red cells into the incubation medium via benzidine hydrochloride, that joins hemoglobin in the presence of H2O2 and stains the solution blue, the intensity of staining being measured spectrophotometrically. The authors consider radiometry preferable; this method makes use of 3H uridine-labeled chicken nuclear red cells as targets, and the results of the test are analyzed ina beta-scintillator. Less patient's blood is necessary for this test, since lower concentrations of effector cells may be used.