Latent Form Research Articles

Relationshipof clinical course, biochemical and immunological changes in patients with chronic hepatitis B+C

Recent studies have shown that parenteral viral hepatitis is one of the main causes of chronic diffuse liver diseases. The number of patients not only with chronic hepatitis C and B, but also patients with hepatitis of mixed etiology is increasing annually. At the same time, there is a potentiating effect of the association of HBV and HCV with a high risk of developing progressive forms of chronic viral hepatitis or hepatocellular carcinoma. Correlation of the clinical course, biochemical indices in patients with viral mixed infection of the liver have not been studied enough. In the studies of a number of authors, it has been shown that with mixed infection, erased and latent forms of the disease are rarely observed. The aim of the study is to study the immune status and biochemical parameters in patients with chronic hepatitis B+C for a more accurate assessment of the severity of the pathological process. Materials and methods. 62 patients with chronic hepatitis B+C were examined. All the patients under examination were monitored in the hepatological center of the Odessa Municipal Infectious Hospital (Ukraine). The assessment of the severity of the patients' condition was carried out using biochemical indicators. Confirmation of the diagnosis based on determination (qualitatively and quantitatively) HCV RNA and HBV DNA by polymerase chain reaction. Subpopulations of B- and T-lymphocytes (CD19+, CD16+, CD8+, CD4+, CD3+) were determined by the immunofluorescence method using a set of polyclonal and monoclonal antibodies to establish differential antigens of human lymphocytes using the Eurostar immunofluorescence microscope. Results and discussions. When analyzing the main clinical syndromes in dynamics, it was found that in patients with chronic hepatitis B+C with initial of fibrosis (F0-F1), less pronounced changes in the general condition were observed than in patients with moderate (F2) and advanced fibrosis (F3). The presence of a correlation between the degree of fibrosis and immunological indices in patients with chronic hepatitis B+C allows us to use the information obtained as one of the additional criteria for the severity of the pathological process. The expressiveness of changes of immune status indicators can serve as an additional criterion for the degree of morphological abnormalities in liver tissue.

Alpha2 Adrenergic Modulation of Spike-Wave Epilepsy: Experimental Study of Pro-Epileptic and Sedative Effects of Dexmedetomidine.

In the present report, we evaluated adrenergic mechanisms of generalized spike-wave epileptic discharges (SWDs), which are the encephalographic hallmarks of idiopathic generalized epilepsies. SWDs link to a hyper-synchronization in the thalamocortical neuronal activity. We unclosed some alpha2-adrenergic mechanisms of sedation and provocation of SWDs in rats with spontaneous spike-wave epilepsy (WAG/Rij and Wistar) and in control non-epileptic rats (NEW) of both sexes. Dexmedetomidine (Dex) was a highly selective alpha-2 agonist (0.003-0.049 mg/kg, i.p.). Injections of Dex did not elicit de novo SWDs in non-epileptic rats. Dex can be used to disclose the latent form of spike-wave epilepsy. Subjects with long-lasting SWDs at baseline were at high risk of absence status after activation of alpha2- adrenergic receptors. We create the concept of alpha1- and alpha2-ARs regulation of SWDs via modulation of thalamocortical network activity. Dex induced the specific abnormal state favorable for SWDs-"alpha2 wakefulness". Dex is regularly used in clinical practice. EEG examination in patients using low doses of Dex might help to diagnose the latent forms of absence epilepsy (or pathology of cortico-thalamo-cortical circuitry).

Extracellular Vesicle-associated GARP/TGFβ:LAP Mediates "Infectious" Allo-tolerance.

C57BL/6 mice were tolerized by i.p. injection of CBA/J splenocytes followed by anti-CD40L/CD154 antibody treatment on days 0, 2, and 4. On day 35, spleen and lymph nodes were extracted and isolated lymphocytes were restimulated with sonicates of CBA splenocytes overnight. sEVs were extracted from culture supernatants by ultracentrifugation (100 000g) and assayed for (a) the presence of TGFβ:LAP associated with tetraspanins CD81,CD63, and CD9 by enzyme-linked immunosorbent assay; (b) GARP, critical to membrane association of TGFβ:LAP and to activation from its latent form, as well as various TGFβ receptors; and (c) TGFβ-dependent function in 1° and 2° immunosuppression of tetanus toxoid-immunized B6 splenocytes using trans-vivo delayed-type hypersensitivity assay. After tolerization, CBA-restimulated lymphocytes secreted GARP/TGFβ:LAP-coated extracellular vesicles. Like IL35 subunits, but unlike IL10, which was absent from ultracentrifuge pellets, GARP/TGFβ:LAP was mainly associated with CD81+ exosomes. sEV-bound GARP/TGFβ:LAP became active in both 1° and 2° immunosuppression, the latter requiring sEV uptake by "bystander" T cells and reexpression on the cell surface. Like other immune-suppressive components of the Treg exosome, which are produced in a latent form, exosomal GARP/TGFβ:LAP produced by allo-specific regulatory T cells undergoes either immediate activation (1° suppression) or internalization by naive T cells, followed by surface reexpression and subsequent activation (2°), to become suppressive. Our results imply a membrane-associated form of TGFβ:LAP that, like exosomal IL35, can target "bystander" lymphocytes. This new finding implicates exosomal TGFβ:LAP along with Treg-derived GARP as part of the infectious tolerance network.

Thermal exchanges, physiological responses and productive performance of Guinea Fowl (Numidia meleagris) subjected to different air temperatures.

The objective of this research was to evaluate the thermal exchanges, physiological responses, productive performance and carcass yield of Guinea Fowl confined under thermoneutral conditions and under thermal stress. For the experiment, 96 animals were confined in 8 experimental boxes of 1 m2 of area, each, divided in equal numbers and placed inside two distinct climatic chambers, where the birds were distributed in a completely randomized design, with two treatments (air temperatures of 26 and 32°C, respectively). For the collection of physiological responses and carcass yield 16 birds were evaluated and for the collection of data on feed and water consumption and productive responses, 48 birds per treatment were evaluated. The environmental variables (air temperature (AT), air relative humidity and wind speed), temperature and humidity index (THI), heat exchanges, physiological responses (respiratory rate, surface temperature, cloacal temperature and eyeball temperature), feed (FC) and water (WC) consumption and production responses (weight gain, feed conversion index and carcass yield) of the birds were evaluated. With the elevation of the AT, it could be noticed that the THI went from a thermal comfort condition to an emergency condition, where the birds lost part of their feathers, increased all physiological responses evaluated, and consequently, reduced by 53.5% the amount of heat dissipated in the sensible form and increased by 82.7% the heat losses in the latent form, increasing also the WC. ATs of up to 32°C did not significantly affect the productive performance and carcass yield of the guinea fowl.

The prognostic significance of the activities of matrix metalloproteinases-2 and -9 in dogs for mammary gland neoplasia (pilot study)

The aim of this study was to determine the activity of matrix metalloproteinase-2 and -9 in blood and tumour tissue of female dogs with benign and malignant mammary gland neoplasias. The increased activity of matrix metalloproteinase-2 and -9 was registered in 3.8–8.9% of canines with benign neoplasias (adenoma, mixed tumour) and in 32.5–63.5% of those with malignant neoplasias (carcinoma: mixed type, ductal, tubular, tubulopapillary). Neoplastic transformation is accompanied by blood level increase of both latent and active enzyme forms. The dogs with malignant mammary gland neoplasias were diagnosed with credible increase of matrix metalloproteinases-2 activity up to 1.59–1.96 in blood plasma and up to 21.57–24.84 in tumour tissue and the increase of matrix metalloproteinases-9 activity up to 2.16–2.67 and 29.53–35.26 respectively. For benign mammary neoplasms, the proportion of dogs with enhanced expression of matrix metalloproteinase-2 was higher than the number of patients in whom an increase in the level of matrix metalloproteinase-9 or both of these enzymes was registered by 1.7 times, for dogs with malignant tumours – in blood plasma – by 1.4–1.6 times, dogs with neoplastic changed functional tissue – 1.7 and 1.9 times, respectively. Histological type and metastatic foci presence did not correlate with enzymes’ activity. The enzymes’ activity figures in benign neoplasias fluctuated within those in clinically healthy animals. A positive characteristic of determining the plasma and tissue expression level of matrix metalloproteinases in dogs with mammary tumours is the low degree of invasiveness of the method against the background of the high informativeness of the results obtained in the preoperative period. The obtained results prove the possibility of using matrix metalloproteinases-2 and -9 to predict the course and to control the treatment of mammary neoplasia.

Modern trends of the incidence of syphilis in the areas of Ural Federal District

BACKGROUND: The incidence of syphilis is an indicator of social disadvantage in society as a whole and in particular regions, and is mostly determined by medical, social, biological and economic factors.
 AIM: To study modern clinical and epidemiological aspects and assess epidemic trends of syphilis in the population of the Ural Federal District for the period from 2014 to 2022 in order to determine further preventive epidemic control measures.
 MATERIALS AND METHODS: The object of the study are the data of statistical forms No. 9 and No. 34 on the number of registered cases of syphilis in the Ural Federal District for the period from 2014 to 2021, the data of statistical materials "Resources and activities of medical organizations of dermatovenerological profile. Incidence of sexually transmitted infections, contagious skin diseases and skin diseases" and operational data for 6 months of 2021 and 2022 years. Intensive and extensive morbidity rates are calculated and their analysis using descriptive statistics is presented.
 RESULTS: The main patterns of the course of syphilitic infection in six subjects (which are parts of the Ural Federal District) of the Russian Federation have been studied. Modern clinical and epidemiological features and trends have been established: variability of morbidity indicators, its clinical structure and territorial dynamics; variability in the age-associated characteristics of patients, the emergence of high-risk groups have been noted. Target indicators characterizing the epidemiological situation of syphilis in the region have been determined. The necessity of interdisciplinary interaction of doctors of various specialties on early detection of patients with syphilis is indicated.
 CONCLUSIONS: During the analyzed period from 2014 to 2022, in certain areas of the Ural Federal District, the current trends of syphilitic infection prevalence is characterized by multi-directional dynamics and levels of morbidity, an increase in early and late forms of syphilis in the structure of clinical forms of the disease; an increase in the registration of late latent forms of infection, cases of congenital syphilis, an increase in the incidence rate among adolescents, foreign citizens and pregnant women in the territory of the Ural Federal District.

Directed differentiation of human iPSCs into mesenchymal lineages by optogenetic control of TGF-β signaling

In tissue development and homeostasis, transforming growth factor (TGF)-β signaling is finely coordinated by latent forms and matrix sequestration. Optogenetics can offer precise and dynamic control of cell signaling. We report the development of an optogenetic human induced pluripotent stem cell system for TGF-β signaling and demonstrate its utility in directing differentiation into the smooth muscle, tenogenic, and chondrogenic lineages. Light-activated TGF-β signaling resulted in expression of differentiation markers at levels close to those in soluble factor-treated cultures, with minimal phototoxicity. In a cartilage-bone model, light-patterned TGF-β gradients allowed the establishment of hyaline-like layer of cartilage tissue at the articular surface while attenuating with depth to enable hypertrophic induction at the osteochondral interface. By selectively activating TGF-β signaling in co-cultures of light-responsive and non-responsive cells, undifferentiated and differentiated cells were simultaneously maintained in a single culture with shared medium. This platform can enable patient-specific and spatiotemporally precise studies of cellular decision making.

Immune recognition of latent neuronal Toxoplasma gondiiinfection

Abstract Toxoplasma gondii is a neurotropic parasite with high mortality rates in immunocompromised individuals. Successful therapies have been developed against the acute tachyzoite stage, but no treatment options exist to target the latent form of the parasite – bradyzoites that form cysts within neurons. It is thought that residence of cysts in neurons shelters T. gondii from immune recognition and clearance. However, a decline in cyst numbers over time suggests the presence of anti-cyst immunity. Additionally, neuronal deletion of STAT1 results in increased cyst size and number, but no change in tachyzoite replication, providing evidence for IFN-γ-mediated cyst control. T cells are a major source of IFN-γ and are crucial for parasite control in the CNS, but it remains unclear if T cells directly target cyst-infected neurons. The use of transgenic parasites with bradyzoite-restricted OVA expression (BagOVA) reveals that cyst-derived antigen can induce OT-I CD8+ T cell responses in the brains of chronically infected mice. These cyst-specific OT-Is have distinct effector phenotypes and cytokine production when compared to T cells responding to antigen expressed by both tachyzoites and bradyzoites (ptubOVA). To determine whether neurons directly present cyst antigen to induce CD8+ T cell responses in BagOVA infection, mice with neuronal-specific loss of MHCI were generated. Infection of these mice showed no significant change in the magnitude of T cell responses or parasite burden in the CNS. Together these findings reveal that T cell responses can be mounted against cyst antigens, but another cell type likely mediates cross presentation of cyst derived antigens. Supported by grants from NIH (R01 580-5803-4-580697-XXXX-2000-3000)

CD8 +T cell recognition of neuronal cysts during chronic Toxoplasma gondiiinfection

Abstract The parasite Toxoplasma gondiiestablishes persistent infection within neurons by forming cysts that harbor latent form bradyzoites. The ability to form cysts is dependent on the parasite transcription factor BFD1 and, given the presence of a thick cyst wall, it was previously thought that cysts were protected from immune recognition. However, the generation of a BAG1-OVA parasite line, in which OVA protein expression is restricted to bradyzoites, revealed that in vivo infection elicited the activation and expansion of OVA-specific OT-I CD8 +T cells. To determine whether endogenous CD8 +T cells can recognize natural cyst-derived antigens and potentially participate in cyst control, in vitro T cell recognition assays were performed using a BFD1-inducible line of parasites that form in vitro cysts. In these assays, CD8 +T cells isolated from T. gondii-infected mice recognized both infected murine fibroblasts and neurons containing cysts. These studies indicate that cyst-specific T cells are present during infection and future studies will explore if any parasite antigens transported beyond the cyst wall can be presented by neurons for T cell recognition. Supported by NIH grant R01 AI157247-01

Lived experiences of people living with HIV-A qualitative exploration on the manifestation, drivers, and effects of internalized HIV stigma within the Malawian context.

HIV-related internalized stigma remains a major contributor to challenges experienced when accessing and providing HIV diagnosis, care and treatment services. It is a key barrier to effective prevention, treatment and care programs. This study investigated experiences of internalized stigma among people living with HIV in Malawi. A participatory cross-sectional study design of participants from eight districts across the three administrative regions of Malawi. Data were collected using Key Informant Interviews (n = 22), Focus Group Discussions (n = 4) and life-stories (n = 10). NVIVO 12 software was used for coding applying both deductive and inductive techniques. Health Stigma and Discrimination Framework was used as a theoretical and analytical framework during data analysis. Overt forms of stigma and discrimination were more recognizable to people living with HIV while latent forms, including internalized stigma, remained less identifiable and with limited approaches for mitigation. In this context, manifest forms of HIV-related stigma intersected with latent forms of stigma as people living with HIV often experienced both forms of stigma concurrently. The youths, HIV mixed-status couples and individuals newly initiated on ART were more susceptible to internalized stigma due to their lack of coping mechanism, unavailability of mitigation structures, and lack of information. Broadly, people living with HIV found it difficult to identify and describe internalized stigma and this affected their ability to recognize it and determine an appropriate course of action to deal with it. Understanding the experiences of internalized stigma is key to developing targeted and context specific innovative solutions to this health problem.

Semi-autonomous wound invasion via matrix-deposited, haptotactic cues

Proper wound healing relies on invasion of fibroblasts via directed migration. While the related experimental and mathematical modeling literature has mainly focused on cell migration directed by soluble cues (chemotaxis), there is ample evidence that fibroblast migration is also directed by insoluble, matrix-bound cues (haptotaxis). Furthermore, numerous studies indicate that fibronectin (FN), a haptotactic ligand for fibroblasts, is present and dynamic in the provisional matrix throughout the proliferative phase of wound healing. In the present work, we show the plausibility of a hypothesis that fibroblasts themselves form and maintain haptotactic gradients in a semi-autonomous fashion. As a precursor to this, we examine the positive control scenario where FN is pre-deposited in the wound matrix, and fibroblasts maintain haptotaxis by removing FN at an appropriate rate. After developing conceptual and quantitative understanding of this scenario, we consider two cases in which fibroblasts activate the latent form of a matrix-loaded cytokine, TGFβ, which upregulates the fibroblasts’ own secretion of FN. In the first of these, the latent cytokine is pre-patterned and released by the fibroblasts. In the second, fibroblasts in the wound produce the latent TGFβ, with the presence of the wound providing the only instruction. In all cases, wound invasion is more effective than a negative control model with haptotaxis disabled; however, there is a trade-off between the degree of fibroblast autonomy and the rate of invasion.

Diphyllobothriasis: diagnosis, treatment, prophylaxis

Helminthiases are found in the group of the most common and socially significant human infectious diseases. According to current data, at least 1 billion people are infected with helminths or pathogenic infections. One of the most important and widespread human biohelminth parasites in the Russian Federation is the broad-spectrum tapeworm (Diphyllobothrium latum). The absence of specific clinical diagnosis, as well as the predominance of latent forms, determines the presence of diphyllobothriasis disease. The aim of the study was to summarize current data on the epidemiology, pathogenesis, clinical and laboratory manifestations and therapy of diphyllobothriasis. Materials and methods. Clinical cases of diphyllobothriasis were analyzed among patients who were treated in January to November 2022 in the departments of the daytime period of clinical observation of the infectious diseases hospital named after S.P. Botkin, as well as the department of incidents detected and the clinical diagnostic center of Pediatric Research and Clinical Center for Infectious Diseases. Cases of the use of diseases, data from laboratory and instrumental studies, and therapy were evaluated. The most informative of them is given as a clinical example. The article also describes in detail the current biological and epidemiological features of Diphyllobothrium latum, the characteristic clinical picture of diseases and the most frequent complications. Results. Diphyllobothriasis still remains one of the most vulnerable biohelminthiases in Russia, typical for some patients of different ages. Verification of the diagnosis of “diphyllobothriasis” can be of the nature of an “accidental finding” during screening scatological or instrumental studies; upon admission, patients themselves begin to suspect helminthiasis when fragments of the strobila worm are found in the feces. To identify a conditional diagnosis of the pattern of clinical and laboratory infection (exclusion of a complicated course of diseases, helminthiases of a different etiology) and to determine risk factors for the transmission of diseases for the prevention of infections.

Tuberculosis Among Lemurs’ Captive Breeding at the Parc Botanique et Zoologique de Tsimbazaza (PBZT) in the Madagascar

The Madagascar is a country with a high endemic biodiversity. For example, it homes 112 species and subspecies of lemurs with 100% of endemism. They are endangered in their natural habitat due to different threats from anthropogenic pressure and climate change. Between May to September 2022, eight relatives <i>Varecia varecia substincta</i> captive breeding at the Parc Botanique et Zoologique de Tsimbazaza deceased in serial and unexpectedly, some of them by tuberculosis. This study is carried out to detect the reasons why these lemurs are died and then, to assess the tuberculosis prevalence among the remain alive lemurs, in objective to proceed in cure. Tuberculosis is a notifiable treatment in the Madagascar. Outcomes has shown that lemurs are predisposed to tuberculosis infection as human primates. It may be present in latent or fatal active forms as miliary tuberculosis. Vulnerabilities depend on gender and species. Females are more fragile than males. Among different species of lemurs,<i> Varecia variegata subtincta</i> are the most threatened to tuberculosis followed by <i>Eulemur fulvus</i>. Tuberculosis is hereditary and for the first time, its genetic aspects are discovered in the Madagascar. Foremost, genetic predisposition has an influence in individual immune responses. In fact, change in weathers factors has increased underlying individual susceptibilities which led to a serious health problem. Consanguinities are an addition threat for tuberculosis transmission in an endemic island context. These basic fields need further confirmation both for lemurs’ conservation and to sustain the control of tuberculosis in the Madagascar.

EFFECTS OF LATENT TOXOPLASMOSIS ON AUTOIMMUNE THYROID DISEASES IN PREGNANT WOMEN - DUHOK CITY-KURDISTAN REGION- IRAQ

Background: Toxoplasmosis is a common zoonotic disease has a global distribution and can infect many hosts causes several clinical symptoms in humans and attack many body organs leading to hormonal and behavioral alterations in infected hosts. Latent or dormant form toxoplasmosis is the common form that can affect pregnancy course. Autoimmune thyroid disease (AITD) belongs to the known risk factors for adverse pregnancy outcomes. The aim of this study was the detection of anti-Toxoplasma antibodies and its effects on autoimmune thyroid disease among pregnant women.

Immunological characterization of a VIR protein family member (VIR-14) in Plasmodium vivax-infected subjects from different epidemiological regions in Africa and South America.

Plasmodium vivax is a major challenge for malaria control due to its wide geographic distribution, high frequency of submicroscopic infections, and ability to induce relapses due to the latent forms present in the liver (hypnozoites). Deepening our knowledge of parasite biology and its molecular components is key to develop new tools for malaria control and elimination. This study aims to investigate and characterize a P. vivax protein (PvVir14) for its role in parasite biology and its interactions with the immune system. We collected sera or plasma from P.vivax-infected subjects in Brazil (n = 121) and Cambodia (n = 55), and from P. falciparum-infected subjects in Mali (n = 28), to assess antibody recognition of PvVir14. Circulating antibodies against PvVir14 appeared in 61% and 34.5% of subjects from Brazil and Cambodia, respectively, versus none (0%) of the P. falciparum-infected subjects from Mali who have no exposure to P. vivax. IgG1 and IgG3 most frequently contributed to anti-PvVir14 responses. PvVir14 antibody levels correlated with those against other well-characterized sporozoite/liver (PvCSP) and blood stage (PvDBP-RII) antigens, which were recognized by 7.6% and 42% of Brazilians, respectively. Concerning the cellular immune profiling of Brazilian subjects, PvVir14 seroreactive individuals displayed significantly higher levels of circulating atypical (CD21- CD27-) B cells, raising the possibility that atypical B cells may be contribute to the PvVir14 antibody response. When analyzed at a single-cell level, the B cell receptor gene hIGHV3-23 was only seen in subjects with active P.vivax infection where it comprised 20% of V gene usage. Among T cells, CD4+ and CD8+ levels differed (lower and higher, respectively) between subjects with versus without antibodies to PvVir14, while NKT cell levels were higher in those without antibodies. Specific B cell subsets, anti-PvVir14 circulating antibodies, and NKT cell levels declined after treatment of P. vivax. This study provides the immunological characterization of PvVir14, a unique P. vivax protein, and possible association with acute host's immune responses, providing new information of specific host-parasite interaction. Trial registration: TrialClinicalTrials.gov Identifier: NCT00663546 & ClinicalTrials.gov NCT02334462.

Abstract 6326: Discovery &amp; preclinical evaluation of BPB-101: A novel triple functional bi-specific antibody targeting GARP-TGFβ complex/SLC, free TGF-β and PD-L1

Abstract Background: Recent studies on tumor microenvironment (TME) have been conferred considerable attention, especially the transforming growth factor-β (TGF-β) signal axis and the PD-L1/PD-1 signal axis. TGF-β is a crucial enforcer in promoting tumor cells angiogenesis, plasticity, fibrosis, and in regulating immune homeostasis and tolerance. The TGF-β signaling controls innate and adaptive immunity in TME by inhibiting the activity of natural killer (NK) cells and effector T cells, while promoting the expansion of regulatory T cells (Tregs). Many studies have revealed that free TGF-β mainly released from glycoprotein A repetitions predominant (GARP)-TGFβ complex at the surface of activated Tregs, B, NK, and tumor cells. Existing antibodies or fusion proteins targeting TGF-β cannot simultaneously block the TGF-β releasing and naturalized free TGF-β in TME, thus limiting the anti-tumor effect. Herein, we designed a bi-specific antibody named as BPB-101 with triple functions: specifically target GARP-TGFβ complex and/or small latent complex (SLC), free TGF-β, and PD-L1. BPB-101 can stop tumor expansion and recruitment, and may enhance the immune effector cells to eliminate the tumor. Methods: An anti-GARP mAb was discovered using a hybridoma platform and selected for binding with GARP-TGFβ complex/SLC and free TGF-β. An anti-PD-L1 nAb was screened out by VHH library and optimized in tumor-bearing mice. After humanizing, these two antibodies were successfully assembled to form a unique bi-specific antibody, BPB-101. The binding affinity of BPB-101 to GARP-TGFβ complex/SLC, free TGF-β and PD-L1 was determined by ELISA, FACS and SPR/BLI. Report gene assay was utilized to detect the blocking activity of BPB-101. The TGF-β secretion from activated Tregs was also analyzed. Finally, the anti-tumor efficacy of BPB-101 was assessed with a transgenic mouse model (B6-hLRRC32). To address the biosafety risks, the nonclinical safety profile was also evaluated in cynomolgus monkeys. Results and Conclusion: BPB-101 is a high-affinity antibody aims to reverse immunosuppressive TME by triple-targeting functions. The blocking ability of TGF-β in not only latent form but also free form makes our BPB-101 distinct from M7824, any other existing anti-TGF-β mAbs and anti-GARP mAbs. In a MC38-hPD-L1 tumor-bearing B6-hLRRC32 model, BPB-101 showed significant tumor growth inhibition of 93 % at the dose of 5 mg/kg (p&amp;lt;0.001), showing superior efficacy to monoclonal antibody alone or in combination with anti-PD-L1 nAb. In addition, BPB-101 showed durable responsiveness and a splendid safety profile with manageable adverse events. These striking results support the clinical development potential of our trifunctional agent as a monotherapy and in combination with other immunotherapies. Citation Format: Wenxin Xu, Jieying Xu, Deyu Xu, Hongjie Cheng, Li Zhang, Huan Zheng, Siyuan Ye, Mengshi Jiang, Jiabing Wang, Lieming Ding. Discovery &amp; preclinical evaluation of BPB-101: A novel triple functional bi-specific antibody targeting GARP-TGFβ complex/SLC, free TGF-β and PD-L1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6326.

Roles of viable but non-culturable state in the survival of Campylobacter jejuni.

Despite being considered fragile and fastidious, Campylobacter jejuni is the most prevalent cause of foodborne bacterial gastroenteritis, and chicken meat is considered the main vehicle of transmission to humans. This agent can survive adverse conditions in the form of biofilms, but extreme stress (nutritional, oxidative and thermal) promotes the acquisition of a state called viable but not culturable (VBNC). The emergence of this pathogen worldwide and the recent international requirements in its control instigated us to qualitatively and quantitatively estimate the time required for the acquisition of the VBNC form in 27 strains of C. jejuni, characterize morphological aspects, determine its adaptive and invasive potential and perform comparative metabolomic evaluation. Extreme stress promoted the complete acquisition of the VBNC form in a mean time of 26 days. Starting from an average initial count of 7.8 log CFU/mL, the first four days determined the greatest average reduction of the culturable form of 3.2 log CFU/mL. The scanning and transmission image analyses showed a transition from the typical viable form (VT) to the VBNC form, with initial acquisition of the straight rod shape, followed by loss of the flagella and subdivision into two to 11 imperfect cocci arranged in a chain and rich in cellular content, until their individual release. RT-PCR identified the presence of ciaB and p19 transcripts in the 27 cultivable C. jejuni strains, a character maintained in the VBNC form only for p19 and in 59.3% (16/27) of the VBNC strains for the ciaB gene. The average inoculation of 1.8 log CFU/mL of C. jejuni VBNC into primary chicken embryo hepatocyte cells promoted the occurrence of apoptosis processes significantly after 24 hours of contact by one of the strains tested. In C. jejuni VBNC, we detected higher expression of metabolites linked to protective and adaptation mechanisms and of volatile organic precursor compounds indicative of metabolism interruption. The oscillations in the time of acquisition of the VBNC form together with the presence of transcripts for ciaB and p19, the identification of cell lysis and metabolites that ensure the maintenance of the pathogen alert to the fact that C. jejuni VBNC remains virulent and adapted to stress, which makes evident the potential danger of this latent form, which is not detectable by official methodologies.

Application of BI-V for contact children and adolescents from focies of multidrug-resistant tuberculous infection: evaluation of effectiveness

Objective — to study the need of using the natural immunomodulator ВІ-V for contact children and adolescents from focies of multidrug-resistant tuberculous infection (MDR-TBI) for a non-specific immunoprevention of tuberculosis (TB). Materials and methods. A retrospective analysis of the medical records of 120 contacts (75 children and 45 adolescents) from focies of MDR-TBI and 102 adult with TB, who became the source of infection was carried out. Microbiological examination in adults included the detection of Mycobacterium tuberculosis (MBT) in sputum by smear microscopy, sowing of material on Levenstein—Jensen medium, typing of isolated MBT on BACTEC MGIT 960, determination of the drug susceptibility test (DST) of MBT to antimycobacterial drugs (AMBDs) of first- and second-line, as well as molecular genetic researched of sputum, in particular by the method of GeneXpert MTB/RIF and linear probe analysis. In the contact children from focies of MDR-TBI the determination of the population and subpopulation composition of blood lymphocytes was carried out among 25 children before and among 22 children after immuno­preven­tion. Results and discussion. Most contact children and adolescents (61.7 %) from focies of MDR-TBI had concomitant diseases. The prophylactic administration of the BI-V immunomodulator reduced the frequency of acute respiratory viral infections and exacerbations of non-specific bronchopulmonary diseases by 2.0 times, the development of an active process from the latent tuberculous infection by 2.6 times, the frequency of transformation of latent tuberculosis infection into an active process by 2.6 times, improved appetite, memory, learning outcomes and increased physical activity. The latent tuberculous infection based on the Mantoux test results was found in 82.5 % (99) children. The quantiferon test confirmed the presence of a latent form of tuberculosis in 104 (86.7 %) contacts, the rest of the test results were negative. In children who received ВІ-V, latent tuberculous infection turned into an active process 2.8 times less often than in infected children who did not take this immunomodulator. The existing deviations into the regulatory system and the cellular response system disappeared after the autumn-spring course of ВІ-V. Conclusions. Immunomodulator ВІ-V is an effective preventive means for bronchopulmonary diseases, including MDR-TB among contact children and adolescents from focies of MDR-TBI.

The influence of the Epstein-Barr virus on paraclinical indicators and cytokine levels in children with rotavirus gastroenteritis

The aim. To identify the influence of the latent form of Epstein-Barr virus infection on the clinical picture, parameters of clinical blood analysis and cytokine response of children with rotavirus gastroenteritis.&#x0D; Materials and methods. 56 children were examined. Of them, 33 children (group 1) had no background infection with herpesviruses, and 23 patients (group 2) suffered from rotavirus gastroenteritis against the background of Epstein-Barr virus (EBV) infection. The patients were comparable in terms of sex, age and disease severity. Differences at p &lt;0.05 were considered statistically significant.&#x0D; Results. The analysis of the obtained data showed that the latent form of EBV infection affects the clinical picture, indicators of clinical blood analysis and the reaction of serum cytokines of children with rotavirus gastroenteritis.&#x0D; Сonclusions. A comparative analysis revealed that a latent form of herpesvirus infection in children with rotavirus gastroenteritis is associated with lower temperature reaction and a lower frequency of vomiting in the acute period of the disease against the background of longer persistence of fever, diarrhoea and catarrhal syndromes. On the part of the investigated laboratory indicators, the presence of EBV infection is characterized by a lower level of haemoglobin (р = 0,013) against a higher content of neutrophils (p = 0.002), eosinophils (p = 0.033) and monocytes (p &lt; 0.001) in the acute period of RVI, and a significantly higher content of eosinophils (p = 0.001) and monocytes (p &lt; 0.001) against the background of a lower level of the relative content of lymphocytes compared to patients of Group 1 in the period of early convalescence. Changes in the cytokine response in children with mixed infection are characterized by a significantly lower concentration of IL-1β at the onset of RVI and a higher content of IL-4 and TNF-α throughout the disease compared to children with mono-RVI

Cell-mediated and humoral immunity during COVID-19 in the Republic of Crimea

The COVID-19 (coronavirus disease 2019) pandemic has spurred the development of highly effective quantitative methods for assessing the adaptive immune response to the SARS-CoV-2 (severe acute respiratory syndrome-related coronavirus 2) virus. In order to assess the humoral component of the immune response, various methods for detecting immunoglobulins A, M, G are widely used. ELISPOT seems to be the most accessible and effective method to assess the level of T cells that specifically respond to the SARS-CoV-2 virus antigens.The aim. To assess cell-mediated and humoral immunity in COVID-19 in residents of the Republic of Crimea.Methods. The study was performed on 24 volunteers: the presence of coronavirus antibodies was determined by ELISA method, and the presence of contact with coronavirus proteins – by the ELISPOT “TigraTest® SARS-CoV-2” method (Generium, Russia). For retrospective study of humoral immunity in the population, we assessed 10 000 ELISA tests (ECOlab IgM and IgG, Russia) performed in our laboratory for the period from July 2020 to January 2022.Results. The results show the effectiveness of using the ELISPOT method to detect latent forms of coronavirus infection. It is important to note that there is statistically significant relationship between the timing of the disease and the number of spots in both antigen panels. After vaccination against SARS-CoV-2, cell-mediated immunity lasts up to 6 months or more.Conclusions. As a result of the study, it was found that during 2021, the level of immunization of the population of the Republic of Crimea against COVID-19 has significantly increased; the proportion of residents who have positive IgG test has increased from 27 % to 87 %. The results of ELISPOT studies using a set of reagents for in vitro detection of blood T-lymphocytes that specifically respond to SARS-COV-2 virus antigens (“TigraTest® SARS-CoV-2”) showed that this method is more sensitive than ELISA in detecting latent diseases.