CD4 Lymphocyte Research Articles

Case report: Systemic granulomatous disease with vasculitis in a bull due to hairy vetch (Vicia villosa) toxicosis

A 5-year-old Angus bull presented for generalized alopecia, thickened pleated skin and profuse malodorous watery diarrhea. This bull, along with 6 other beef bulls, had been grazing in a pasture containing hairy vetch (Vicia villosa) for 5-6 months. Diagnostic samples included blood for CBC and chemistry, skin biopsy and scrapings, fecal sample and liver biopsy. Biopsy of skin revealed a chronic, moderate, perivascular and perifollicular, eosinophilic to histiocytic-lymphocytic dermatitis with moderate acanthosis and hyperkeratosis indicative of an ongoing hypersensitivity reaction. The bull did not respond to treatment and was euthanized due to poor prognosis. At necropsy, disseminated systemic granulomatous disease with variable numbers of multinucleated giant cells and eosinophils was observed in multiple organs indicative of systemic granulomatous disease, along with vasculitis in the kidney, liver, and adrenal gland, indicative that vasculitis is involved in the pathogenesis of the lesions. Viral diseases were ruled out by immunohistochemistry of sections of kidney and by in situ hybridization on kidney sections. CD3, CD20, and CD79a lymphocyte markers immunostaining of sections of kidney revealed predominance of CD3 positive lymphocytes, indicative of predominance of T-cell lymphocytes supporting a type-IV hypersensitivity. The vasculitis observed prompted formulating a list of differential diagnoses for infectious diseases that have evidence of vasculitis, including bovine virus diarrhea virus and malignant catarrhal fever. Vasculitis, although previously described in cases of hairy vetch poisoning, is not observed in every single case of the disease. The remaining bulls were not removed from the pasture and none developed clinical signs.

Evaluation of peripheral and central olfactory pathways in HIV-infected patients by MRI

To investigate peripheral and central olfactory pathways using cranial magnetic resonance imaging (MRI) in human immunodeficiency virus (HIV)-infected patients. The cranial MRI images of 37 HIV-infected adult patients and 37 adults without HIV infection having normal cranial MRI results were included in the study. In both groups, olfactory bulb (OB) volume and olfactory sulcus (OS) depth; and insular gyrus and corpus amygdala areas were measured using cranial MRI. In the HIV group, disease duration, HIV RNA, and CD4 lymphocyte count and levels as a percentage were also recorded. The HIV group had significantly lower bilateral OB volumes, insular gyrus and corpus amygdala areas compared to the control group. The HIV group showed positive correlations between OB volumes, OS depths, insular gyrus, and corpus amygdala areas bilaterally. Increases in OB volumes and OS depths were associated with an increase in the insular gyrus area. The corpus amygdala and insular gyrus areas increased similarly. There was no significant correlation between age, gender, disease duration, CD4 lymphocyte count and per cent, HIV RNA values, and the measurement values of the central and peripheral olfactory regions. A decrease in olfactory regions of OB, insular gyrus, and corpus amygdala in HIV-infected patients shows that HIV infection may cause olfactory impairment. There is no correlation between disease duration and olfactory impairment. It may be related to neuroinflammation, HIV-related brain atrophy, acquired immunodeficiency syndrome (AIDS) dementia complex, or neurocognitive impairment, which are the other explanations for the olfactory impairment in HIV. The possible toxicity from antiretroviral therapy (ART) may be another cause that should be investigated further.

Immune response in neuroinflammatory caused by mild traumatic brain injury (experimental study)

The study of the organism immune response in mild traumatic brain injury (mTBI) is an important stage in the formation of a scientific understanding of this type of brain damage. The purpose of the study: to research of the systemic immune response parameters in the experimental modeling of mTBI. The cellular composition and phenotype of immune cell subpopulations were assessed, and the content of pro- and anti-inflammatory cytokines in the mTBI and blood serum of injured rats was determined. A high content of leukocytes and interleukin 6 (IL 6) in the blood was found 2 hours after the injury. The number of cytotoxic T-lymphocytes and B-lymphocytes increased with a decrease of the neutrophils, monocytes, eosinophils and T helpers number one day after mTBI. During the indicated periods of observation, an increase in the content of cytokines TNFα and IL 6 in the cerebrospinal fluid and blood serum was observed, and the indicators of anti-inflammatory IL 10 increased 7–14 days after mTBI. On the 14th day after injury, the level of stab neutrophils remained low, while the number of CD45, CD3, CD20, and CD8 positive lymphocytes continued to increase, which indicates the progression of the inflammatory response. Thus, after the application of mTBI, an inflammatory reaction develops, accompanied by the manifestation of the activity of immune components. Cytokines are registered in the peripheral circulation and cerebrospinal fluid, the number of B-lymphocytes and cytotoxic T-lymphocytes increases.

Risk Factors for CD19-Targeting CAR T Manufacturing Failure and Patient Outcomes: A Report from the UK National CAR T Panel

CAR T manufacturing failure (MF) rates reported in literature range between 1-13%. In some cases the manufacturing process fails to yield a product and in others results in one which does not meet pre-specified criteria for release and labelled an out of specification (OOS) product. We conducted a multicentre retrospective review of factors contributing to MF and its impact on patient outcomes. Patients with relapsed or refractory large B cell lymphoma (LBCL), mantle cell lymphoma (MCL) or B acute lymphoblastic leukemia (B-ALL) approved for CAR T therapy with axicabtagene ciloleucel (axi-cel), brexucabtagene autoleucel (brexu-cel) or tisagenlecleucel (tisa-cel) by the UK national CAR T panel between January 2019 and January 2023 were eligible. Patients with MF were identified from databases of the 11 participating CAR T centres and from the national OOS CAR T panel. In total 1138 patients were approved for CAR T therapy (903 LBCL, 97 MCL and 138 B-ALL). We identified 58 patients who had at least 1 CAR T MF; 38 LBCL (LBCL-MF), 14 MCL and 6 B-ALL with inferred MF frequency of 4.20%, 14.43% and 4.34% respectively. Intended CAR T product was axi-cel in 28 and tisa-cel in 10 LBCL, brexu-cel for all MCL and tisa-cel for all B-ALL patients. Most frequent reasons for MF were poor growth in culture (n=12), low cell viability (n=12) and low T cell purity (n=6). Of the 58 patients with MF, OOS product was available and infused in 19 patients (OOS cohort: 14 LBCL, 2 MCL and 3 B-ALL). Repeat apheresis led to infusion of a product in-specification in 17 patients (MF-delayed-infusion cohort: 10 LBCL, 6 MCL and 1 B-ALL). Another 22 patients did not proceed to CAR T infusion (MF-no-infusion cohort: 14 LBCL, 6 MCL and 2 B-ALL), some despite attempts at remanufacture (n=9) or available OOS product (n=5). We included 25 randomly selected matched (for disease subtype and product) controls (all LBCL: 19 axi-cel and 6 tisa-cel) without MF, 18 of whom received infusion. Univariable analysis of potential baseline clinical factors contributing to risk of MF did not reveal any significant impact of the number of lines of prior therapy, history of prior stem cell transplant or a need for holding therapy prior to apheresis. Bendamustine therapy was significantly associated with risk of MF for all lymphoma patients and the LBCL subset; prior exposure in 4% controls vs 32.7% of all lymphoma MF (p=0.004) and 26.3% of LBCL-MF (p=0.039). This was largely due to therapy within 6 months; 0% of controls vs 19.2% of all lymphoma MF (p=0.026) and 23.7% of LBCL-MF (p=0.009). Factors at apheresis including white cell count, neutrophil count, platelet count, absolute lymphocyte count, CD3 count, CRP, LDH and volume of blood processed had no impact on the risk of MF. With a median follow up of 11.6 months, the 12-month overall survival (OS) from date of approval for CAR T therapy was 45.0% (31.4 - 57.7) for all MF patients (n=58), 49% (24.7 - 69.5) for the OOS cohort (n=19), 64.2% (33.6 - 83.5) for the MF-delayed-infusion cohort (n=17) and 27.3% (11.1 - 46.4) for the MF-no-infusion cohort (n=22). Corresponding 12-month OS was 43.0% (26.7 - 58.3) for all LBCL-MF (n=38), 49.0% (21.6 - 71.7) for LBCL-OOS (n=14), 53.3% (17.3 - 79.8) for the LBCL-delayed-infusion cohort (n=10) and 28.6% (8.8 - 52.4) for the no-infusion cohort (n=14). The 12-month OS for LBCL controls (n=25) was 51.2% (30.3-68.7) for all and 65.5% (38.6 - 82.8) for infused patients (n=18). There was no significant difference between OS for LBCL-MF and LBCL controls (HR 1.38, p=0.31). Though OS was numerically lower for LBCL-OOS compared to infused LBCL controls, it was not statistically different (HR 1.82, p=0.21). For infused patients, the 3 months complete response (CR) rates were similar for the LBCL-OOS (38.5%), LBCL controls (40%) and MF-delayed infusion (53.3%) cohorts. The 3 months overall response rate (OR) was non-significantly lower in the LBCL-OOS (38.5%) compared to LBCL controls (60%) and MF-delayed infusion (53.3%) cohorts. There was no difference between any grade or grade ≥3 CRS or ICANS between the LBCL-OOS, LBCL-control and MF-delayed-infusion cohorts. In summary, outcomes for patients infused with an in-specification or OOS CAR T product following MF are comparable to those of controls. Prior bendamustine therapy within 6 months of apheresis is the only risk factor associated with risk of MF in lymphoma patients. Further analysis will be presented at the meeting.

Factors Influencing Lymphocyte Collection Efficiency for the Manufacture of CAR-T Cells in Adults with B Cell Malignancies , a Single Center Experience

Aim: During leukapheresis, accurate estimation of the proportion of T cells that are removed from peripheral blood (collection efficiency, CE) is essential to minimise apheresis time, reduce lab storage requirements and prevent excess product collection. Based on historical precedent from autologous HPC donors, many centres estimate a CE of 40% for autologous T cell donors, although data are lacking. We aimed to identify patient characteristics and collection parameters that might influence leukapheresis CE in order to maximise efficiency and minimize waste during CAR T manufacture. Methods: We reviewed data from 33 consecutive patients who underwent leukapheresis for CAR-T cell treatment at our centre from March 2020 to October 2022. Apheresis was performed on the Spectra Optia, using the CMNC collection protocol. Apheresis were performed on the Spectra Optia, using the CMNC collection protocol. peripheral collection was used if patients had adequate venous access, otherwise central venous access was used. Peripheral blood flow cytometry was done prior to apheresis. Results: 33 apheresis procedures were documented from patients with 3% acute lymphocytic leukemia (ALL) (n=1), 82% non-Hodgkin lymphoma (NHL) (n=27) and 15% Multiple myeloma (n=5). Median age was 62 (18 -76 ) , Male to Female ratio was 1.38 , the median weight was 75 kg (45-136) the median of prior lines of therapy was 2 (1-7) with 33% of pts receiving prior autologous HCT and 3% Prior allogeneic HCT. The median time from diagnosis to collection was 15 months (3-109) The median WBC value was 6× 10 9/L (2-24), median Hematocrit was 0,34% (0,25-0,45), Median Neutrophil was 4,43×10 9/L (1-21), Monocytes 0,62 ×10 9/L (0,36-1,43), Platelets 185× 10 9/L (94-385), and median absolute Lymphocytes count was 0.8 (0.21-2,4). With median CD3 value was 560×10 9/L and a median percentage of ALC of 76%. CD4 and CD8 absolute median values were 276 and 309 respectively. The median CRP, LDH and Ferritinemia were 29,238 and 252 respectively. The collection was successful in all patients with 32 (97%) having a high CE of >40%. The median CD3 lymphocyte count in the apheresis product was 4,3 × 10 9 with an overall viability of 99% . The median collection efficiency was 71% (31-96) . Median Run time was 210 minutes, total blood volume and apheresis product volume were 9750 and 278ml. In the apheresis product, median CD3 value was 15415 ×10 6 and the median CD3 value per kg was 58×10 6 /kg. In multivariate analysis, baseline characteristics did not significantly impact the collection efficiency. ALC and CD3 pre-apheresis strongly predicted the CD3 yield after apheresis (p< 0,001). No major adverse events were recorded. Conclusion: High efficiency leukapheresis is safe and feasible in autologous T-cell donors. Our data also confirms that high ALC and peripheral blood CD3 counts pre-apheresis are strongly correlated to the CD3 yield in the product. Our centre now uses a CE estimate of 60% for collection of CAR T starting material.

Effect of Plasmodium Infections on CD4 Cells, Neutrophil and Lymphocytes

Knowledge of malaria-immunity is essential for understanding the pathology, treatment & vaccine production. So, changes in CD4 in malaria patients were studied at a teaching hospital in Enugu, Nigeria. 45 patients on doctor’s provisional diagnosis of malaria were examined for Plasmodium infections and the degree of parasitaemia (0/+/ ++/ & +++). Positive/(+) and negative/(0) samples were thereafter counted for CD4 using Flow Cytometry. CD4 cell count of Plasmodium-negative specimens were within reference ranges (RR): (464-1308); mean value: 835. Fifteen (+) parasitaemia showed lower ranges of CD4 count (502-1282); mean: 678; with immune falls in one (427). 9/12 (++) parasitaemia showed crash in CD4 counts, range: 301- 415; mean: 399. Seven (+++) parasitaemia showed crashes in CD4 counts, range: 160-357; mean: 225. As parasitaemia rises, neutrophils, Total WBC and lymphocyte significantly rise in numeric. Malaria causes immunosuppression; unlike prevailing reports, should accelerate complications in immunodeficients-patients, Haemolytic-anaemia results only from chronic cases. CD4 counts should be a paradigm in malaria investigation.

The Predictive Value of CD3+/CD8+ Lymphocyte Infiltration and PD-L1 Expression in Colorectal Cancer.

The immune system plays an important role in tumor development and treatment. In this study, we aimed to determine the relationships among the expressions of PD-L1, CD3, CD8, MMR proteins, clinicopathological features, and prognosis of CRC. Immunohistochemistry was used to determine the expression of PD-L1, CD3, and CD8 in 771 patients with CRC. The expression of PD-L1 in TC was related to the right colon, adenocarcinoma, and dMMR, and in IC, it was related to younger CRC patients and the TNM stage. The expression of CD3 and CD8 in tumor-infiltrating lymphocytes was related to lymph node metastasis and the TNM stage. The expression of PD-L1 in TC and IC was correlated with the infiltration of CD3+ and CD8+ lymphocytes. Univariate survival analysis showed that the expression of PD-L1 in TC, IC, and dMMR was related to a better prognosis. Multivariate survival analysis showed that age, TNM stage, and dMMR were independent prognostic factors for CRC. The OS of the chemotherapy was significantly higher than that of the non-chemotherapy in III-IV TNM stage patients; CRC patients with positive PD-L1 expression in TC or IC and dMMR did not benefit from chemotherapy. PD-L1 expression in TC and IC was closely related to the density of CD3 and CD8 infiltration in tumor-infiltrating lymphocytes. The expression of CD3 and CD8 in tumor-infiltrating lymphocytes and the expression of PD-L1 in IC were linked to the TNM stage of CRC patients. PD-L1 expression in TC and IC and MMR status may act as an important biomarker for guiding the postoperative treatment of III-IV TNM stage CRC patients.

Comparative analysis of clinical and immunological profiles across Omicron BA.5.2 subvariants using next-generation sequencing in a Chinese cohort.

The Omicron BA.5.2 variant of SARS-CoV-2 has undergone several evolutionary adaptations, leading to multiple subvariants. Rapid and accurate characterization of these subvariants is essential for effective treatment, particularly in critically ill patients. This study leverages Next-Generation Sequencing (NGS) to elucidate the clinical and immunological features across different Omicron BA.5.2 subvariants. We enrolled 28 patients infected with the Omicron variant, hospitalized in Zhangjiajie People's Hospital, Hunan, China, between January 20, 2023, and March 31, 2023. Throat swabs were collected upon admission for NGS-based identification of Omicron subvariants. Clinical data, including qSOFA scores and key laboratory tests, were collated. A detailed analysis of lymphocyte subsets was conducted to ascertain the extent of immune cell damage and disease severity. Patients were infected with various Omicron subvariants, including BA.5.2.48, BA.5.2.49, BA.5.2.6, BF.7.14, DY.1, DY.2, DY.3, and DY.4. Despite having 43 identical mutation sites, each subvariant exhibited unique marker mutations. Critically ill patients demonstrated significant depletion in total lymphocyte count, T cells, CD4, CD8, B cells, and NK cells (P < 0.05). However, there were no significant differences in clinical and immunological markers across the subvariants. This study reveals that critically ill patients infected with different Omicron BA.5.2 subvariants experience similar levels of cellular immune dysfunction and inflammatory response. Four mutations - ORF1a:K3353R, ORF1a:L3667F, ORF1b:S997P, S:T883I showed correlation with immunological responses although this conclusion suffers from the small sample size. Our findings underscore the utility of NGS in the comprehensive assessment of infectious diseases, contributing to more effective clinical decision-making.

Comparison of Stress Level, Habitual Physical Activity Level and Immune Profile of People Living With HIV

The objective of this study was to compare the level of stress, the habitual level of Physical Activity and the immunological profile of people living with HIV. The present study had as sample 307 individuals of both sexes, X̅ = 46.8 ± 9.2 years, with positive HIV serology. Socioeconomic information was collected using an instrument, CD4+ lymphocyte count and HIV viral load were collected from the patients' medical records. Stress level was assessed using Lipp's Stress Symptom Inventory (ISSL), and Baecke's questionnaire was used to classify physical activity level. The Man-Whitney test was used to compare the variables and the Spearman test to verify the relationship between the variables adopting p &lt; 0.05 as the significance level. As a result, the study showed that 26.9% of the individuals are in the stress exhaustion phase, in relation to the level of physical activity, 28.2% were identified as "active", 23.1% as "very active", 23.1% as "insufficiently active", and 24.1% as "sedentary". The presence of stress was prevalent in sedentary individuals (13.6%) with longer duration of infection (27.3%) and with lymphocyte counts above 500mm (43.3%). The CD4+ count showed a significant result when compared to the level of stress, (U=1848.50; p=0.02). A relationship between depression/anxiety and stress was identified (r = 0.407; p &gt; 0.001). We conclude that the presence of stress has an effect on TCD4+ count and that the higher the depression/anxiety symptoms the higher the stress level in PLHIV. Keywords: Viral Load, CD4 Lymphocyte Count, Psychological Stress, HIV

POSSIBILITIES OF CORRECTING CELLULAR-HUMORAL IMMUNE DISORDERS AND CYTOKINE STATUS IN HIGH-RISK PREGNANT WOMEN FOR THE DEVELOPMENT OF PREECLAMPSIA

POSSIBILITIES OF CORRECTING CELLULAR-HUMORAL IMMUNE DISORDERS AND CYTOKINE STATUS IN HIGH-RISK PREGNANT WOMEN FOR THE DEVELOPMENT OF PREECLAMPSIA

Role and mechanism of IL-6/STAT3/Th17 signaling pathway in the intervention of forsythia in acute lung injury of septic rats

Objective: To investigate the mechanism of interleukin-6 (IL-6) /signal transducers and activators of transcriptional 3 (STAT3) /helper T cell 17 (Th17) signaling pathway in lung injury of rats with sepsis intervened by forsythia, with a view to provide experimental basis for the role and mechanism of forsythia in the treatment of sepsis. Methods: In July 2021, 30 healthy Wistar male rats were selected and randomly divided into sham operation group, model group and treatment group, with 10 rats in each group. The rat model of sepsis was established by cecal ligation and puncture. 2 h after recovery, the rats were given traditional Chinese medicine forsythia orally, twice a day at an interval of 12 h. The wet/dry mass ratio (W/D) of lung tissue was detected 24 h after surgery. The morphological changes of lung tissue were detected by HE staining. Flow cytometry was used to detect Th17 population in peripheral blood. The expression levels of IL-6 and interleukin-17A (IL-17A) in serum were determined by enzyme-linked immunosorbent assay. The mRNA expression levels of IL-17A and IL-6 in lung tissues were detected by reverse transcription-polymerase chain reaction. The expression levels of STAT3 and IL-17A in lung tissue were determined by Western blotting. Results: Compared with model group, the W/D of lung tissue in treatment group was decreased, and the difference was statistically significant (P<0.05). HE staining of lung tissue showed that compared with the model group, the degree of lung lesion and injury was reduced in the treatment group. Compared with sham operation group, the proportion of Th17 cells in CD4 lymphocytes in peripheral blood of rats in model group was significantly increased (P<0.05). Compared with model group, the proportion of Th17 cells in CD4 lymphocytes in peripheral blood of rats in treatment group was significantly decreased (P<0.05). Compared with sham operation group, peripheral blood serum IL-6 and IL-17 of rats in model group were significantly increased (P<0.05). Compared with model group, IL-6 and IL-17 in peripheral blood serum of rats in treatment group were decreased (P<0.05). Compared with sham operation group, the expressions of IL-17A and IL-6 mRNA in lung tissue of model group were significantly increased (P<0.05). Compared with model group, the expressions of IL-17A and IL-6 mRNA in lung tissue of rats in treatment group were significantly decreased (P<0.05). Compared with sham operation group, the protein expressions of STAT3 and IL-17A in lung tissue in model group were significantly increased (P<0.05). Compared with model group, the pritein expressions of STAT3 and IL-17A in lung tissue in treatment group were significantly decreased (P<0.05) . Conclusion: Forsythia plays a role in alleviating lung injury by down-regulating the expressions of the signaling pathway IL-6/STAT3/Th17, which providing a new target for the treatment of sepsis induced lung injury.

Effect of Nigella sativa on general health and immune system in young healthy volunteers; a randomized, placebo-controlled, double-blinded clinical trial.

Nigella sativa ( N. sativa) is traditionally used as an immune enhancer in different communities. The aim of this study was to evaluate the effect of N. sativa on immunity related parameters in young healthy subjects. This study was a double blind, randomized, placebo controlled clinical trial. Fifty-two healthy subjects (48 male and 4 female) 18-25 years old were enrolled in the study. They were randomly divided into four groups; the first received charcoal capsules and served as controls and the other three received 0.5, 1 g, and 2 g of powdered N. sativa capsules, respectively. Two blood samples were obtained from all participant, before initiation of the trial and at the end of the four weeks intervention. One sample was used for routine health screening by evaluating liver and renal functions as well as complete blood count and differential. The second sample was used to measure certain cytokines including; IL-1, IL-4, IL-6, IL-10, and TNF. A third and fourth samples were obtained from the last cohort of subjects before and after treatment; the third was used for measuring immunoglobulins and CD profile and the fourth for evaluating certain gene expressions (INF-γ, NF-κ-B, TNF-α, IL-1β, IL-13, IL-8, and IL-6). Only 1 g dose of N. sativa produced a significant elevation in total lymphocyte count, CD3+ and CD4+ counts. One gram N. sativa increased the absolute lymphocyte count from 1850±0.24 to 2170±0.26 (p=0.008), CD3+ from 1184.4±75.60 to 1424±114.51 (p=0.009), and CD4+ from 665.6±141.66 to 841±143.36 (p=0.002). This elevation in T cells was lost by increasing the dose of N. sativa to 2g. The rest of the parameters were not changed significantly in all doses. The results show a promising immunopotentiation effect of N. sativa by elevating helper T cells and the optimum dose for young age group seems to be 1 g.

Immunological Responses of Arsenicum album 30CH to Combat COVID-19: Protocol for a Double-Blind, Randomized, Placebo-Controlled Clinical Trial in the Pathanamthitta District of Kerala.

COVID-19 is a recent major public health concern caused by the SARS-CoV-2 virus, with approximately 44.6 million COVID-19-positive cases and 530,000 deaths in India (as of February 1, 2023). The COVID-19 vaccination drive in India was initiated in January 2021; however, an effective preventive strategy with high efficacy and immunological safety remains elusive. The aim of this study is to assess the immunogenic responses of Arsenicum album 30CH (AA30CH) as COVID-19 prophylaxis, including assessment of immunological markers, innate and acquired immune responses, COVID-19 symptoms, and its associated antibody responses. This randomized controlled clinical trial (RCT) will include two parallel comparator groups of AA30CH and placebo with an allocation ratio of 1:1 conducted in the Pathanamthitta district of Kerala, India. The placebo or AA30CH will be administered in three intervention schedules and blood samples will be collected before and after each of the intervention schedules. Based on the inclusion and exclusion criteria, 112 participants per arm (with an expected dropout of 20%) will be screened. Immunogenic responses will be evaluated by determining the antigen density and modulation in immunological markers and lymphocyte subsets CD3, CD4, CD8, CD24, CD27, CD38, CD4 interferon-γ, CD4 CD17, CD4 CD25 (activated T lymphocytes), T cells, B cells, dendritic cells (mature and immature), and natural killer cells on days 1, 5, 23,27, 45, 49, and 66. The innate and acquired immune responses will also be evaluated by a real-time reverse-transcriptase polymerase chain reaction (RT-PCR) array profiler (84-gene set) before and after the study interventions. The toxicity status of AA30CH in study participants will be evaluated through hepatic, renal, and hematological parameters and peripheral smears on days 1, 5, 23, 27, 45, 49, and 66. The number of participants developing COVID-19-like symptoms per National Centre for Disease Control guidelines and the number of participants testing positive for COVID-19 in RT-PCR during follow-ups in any of the three intervention schedules will be identified. Moreover, a subgroup analysis will be used to assess the COVID-19 antibody responses between vaccinated and unvaccinated participants. This RCT protocol has been approved by various committees and funded by the Central Council for Research in Homoeopathy, Ministry of Ayush, Government of India. The project has been implemented in collaboration with the Department of Homoeopathy, Government of Kerala. The RCT was rolled out on January 25, 2023, and enrollment was completed April 3, 2023. The immunological assays will be conducted at the Department of Biotechnology-Translational Health Science and Technology Institute, Faridabad, India. This study will represent the first evaluation of the immunological efficacy and safety of AA30CH in an RCT, which may significantly impact the use of homeopathy as an evidence-based medicine approach. Clinical Trials Registry-India CTRI/2022/08/045089; https://tinyurl.com/mryrpkvk. DERR1-10.2196/48479.

Outcomes of Dolutegravir-Based Antiretroviral Therapy in Thai Youth Aged 18 to 24 Years Living with HIV in Bangkok: A Retrospective Cohort Study

Objective: Dolutegravir (DTG) is recommended globally for people living with HIV. The present study aimed to describe the efficacy and safety of DTG-based regimens among youth aged 18 to 24 years. Materials and Methods: A retrospective cohort study was conducted among youth aged 18 to 24 years living with HIV at Chulalongkorn Hospital. Participants were divided into antiretroviral naïve (AN), antiretroviral experienced with treatment failure (AETF), and antiretroviral experienced and virally suppressed (AEVS). DTG was prescribed as a daily individual tablet or as part of a fixed-dose combination. The primary outcome was virological suppression defined as VL of less than 200 copies/mL within 12 months after DTG initiation. Secondary outcomes included body weight and lipid profile changes. Results: Between 2017 and 2022, 87 youth with a median age of 20.6 (IQR 19.6 to 21.9) years were initiated on DTG. Fifty-six (64.3%) were male. Twelve participants were AN, 18 were AETF, and 57 were AEVS. HIV viral suppression was 100% and 96.2% among those AN and AEVS, but only 54.5% amongst the AETF group (p&lt;0.001). CD4 lymphocyte counts increased in all groups at follow-up. Median body weight change was +0.4 (IQR –1.8 to 3.6) kg in males and +0.5 (IQR –2.8 to 1.7) kg in females. Median total cholesterol declined from 167 (IQR 142 to 186) to 152 (IQR 135 to 170) mg/dL, p=0.003. Median triglycerides declined from 83 (IQR 60 to 129) to 66 (IQR 46 to 78), p&lt;0.001. No hospitalizations or mortalities were observed in the study. Conclusion: DTG was effective and well-tolerated in youth living with HIV, with no significant weight gain and improved lipid profiles. DTG implementation among youth living with HIV should be continued. Keywords: Dolutegravir; Antiretroviral therapy; Adolescents living with HIV

Evaluation of IL-6, FoxP3 Treg lymphocytes, intestinal barrier biomarkers and the use of synbiotics in obese adolescents: a pilot study.

This prospective pilot study evaluated inflammatory and intestinal barrier biomarkers and the effects of a synbiotic in obese adolescents. Eighteen obese and 20 eutrophic adolescents were evaluated for body composition using bioimpedance analysis (BIA), body mass index (BMI), IL-6 and lipopolysaccharide (LPS) serum levels, CD4 and FoxP3 Treg lymphocytes and monocytes. Synbiotic supplementation for 60 days was also evaluated for these parameters only in obese adolescents. We observed an increase in CD4 lymphocyte (18.0 ± 12.4 vs. 8.9 ± 7.5; p < 0.01), IL-6 (0.30 ± 0.06 vs. 0.20 ± 0.06; p = 0.02) and LPS (0.18 ± 0.15 vs. 0.08 ± 0.05; p < 0.01) levels in obese compared to eutrophic adolescents. After synbiotic supplementation, FoxP3 Treg lymphocytes increased (14.0 ± 6.7 vs. 9.9 ± 5.4; p = 0.02) in obese adolescents. Obese adolescents presented a state of microinflammation and intestinal barrier breakdown, and synbiotic supplementation increased the expression of FoxP3 Treg lymphocytes, an anti-inflammatory regulator. Whether the increase in FoxP3 Treg lymphocytes may have an impact on inflammation and outcomes in obese adolescents deserves further evaluation.

Effects of a gut-selective integrin-targeted therapy in male mice exposed to early immune activation, a model for the study of autism spectrum disorder

To clarify the role of gut mucosal immunity in ASD, we evaluated, in the early-life immune activation (EIA) mouse model, the effects of administration of a monoclonal antibody directed against the integrin alpha4 beta7 (α4β7 mAb), blocking the leukocyte homing into the gut mucosa. EIA is a double-hit variant of the maternal immune-activation (MIA) model, including both prenatal (Poly I:C) and postnatal (LPS) immune challenges.In C57BL6/J EIA male adult offspring mice, IL-1β and IL-17A mRNA colonic tissue content increased when compared with controls. Cytofluorimetric analyses of lymphocytes isolated from mesenteric lymph-nodes (MLN) and spleens of EIA mice show increased percentage of total and CD4+α4β7+, unstimulated and stimulated IL-17A+ and stimulated IFN-γ+ lymphocytes in MLN and CD4+α4β7+ unstimulated and stimulated IL-17A+ and stimulated IFN-γ+ lymphocytes in the spleen. Treatment with anti-α4β7 mAb in EIA male mice was associated with colonic tissue IL-1β, and IL-17A mRNA content and percentage of CD4+ IL-17A+ and IFN-γ+ lymphocytes in MLN and spleens comparable to control mice. The anti-α4β7 mAb treatment rescue social novelty deficit showed in the three-chamber test by EIA male mice. Increased levels of IL-6 and IL-1β and decreased CD68 and TGF-β mRNAs were also observed in hippocampus and prefrontal cortex of EIA male mice together with a reduction of BDNF mRNA levels in all brain regions examined. Anti-α4β7 mAb treatment restored the expression of BDNF, TGF-β and CD68 in hippocampus and prefrontal cortex.Improvement of the gut inflammatory status, obtained by a pharmacological agent acting exclusively at gut level, ameliorates some ASD behavioral features and the neuroinflammatory status. Data provide the first preclinical indication for a therapeutic strategy against gut-immune activation in ASD subjects with peripheral increase of gut-derived (α4β7+) lymphocytes expressing IL-17A.

Effectiveness of second-generation integrase strand-transfer inhibitor-based regimens for antiretroviral-experienced people with HIV who had viral rebound.

Antiretroviral regimens containing a second-generation integrase strand-transfer inhibitor (INSTI) plus 2 nucleos(t)ide reverse-transcriptase inhibitors (NRTIs) are the recommended therapy for people with HIV (PWH) who are antiretroviral-naïve or on stable antiretroviral therapy (ART) with viral suppression. Real-world data on the virologic effectiveness of co-formulated bictegravir/emtricitabine/tenofovir alafenamide (BIC/FTC/TAF) among PWH with virologic failure while receiving other ART remain sparse. We retrospectively reviewed the medical records of PWH who had viral rebound with plasma HIV RNA >1000 copies/mL and were switched to either dolutegravir combined with 2 NRTIs or BIC/FTC/TAF. The primary end point was re-achieving viral suppression within the first 48 weeks of switch. The association between NRTI-related resistance-associated mutations (RAMs) and virologic effectiveness was examined. Seventy-nine PWH with viral rebound while receiving other antiretroviral regimens were included. Within the first 48 weeks of switch, the overall probability of re-achieving viral suppression was 79.7% (82.5% [33/40] and 76.9% [30/39] for BIC/FTC/TAF and dolutegravir-based regimens, respectively, p=0.78). PWH with a higher CD4 lymphocyte count (adjusted odds ratio, per 100-cell/mm3 increase, 1.41; 95% confidence interval, 1.02-1.95) were more likely to re-achieve viral suppression. Among PWH switching to BIC/FTC/TAF who had pre-existing RAMs to NRTIs before switch, 14 of 15 (93.3%) successfully achieved viral suppression. Switching to BIC/FTC/TAF and dolutegravir-based regimens could re-achieve viral suppression in four-fifth of the PWH who experienced viral rebound during treatment with other antiretroviral regimens. Pre-existing NRTI-related RAMs did not have adverse impact on the effectiveness of dolutegravir combined with 2 NRTIs or BIC/FTC/TAF.

Immunophenotypic Characterization of Citrate-Containing A Concentrates in Maintenance Hemodialysis: A Pre-Post Study.

Due to chronic inflammation, maintenance hemodialysis (MHD) patients continue to show excess mortality. Acetate-free citrate-buffered A concentrates could be a way to improve the biocompatibility of the procedure, reduce chronic inflammation, and thus in the long term improve the prognosis of patients. Using a pre-post design (3 months of acetate followed by 3 months of citrate-acidified A concentrates in standard bicarbonate-based dialysate hemodialysis, CiaHD) and linear mixed model analysis in 61 stable HD patients, we assessed the impact of CiaHD on counts and phenotypes of peripheral T cells and monocytes by flow cytometry. Switching to CiaHD left C-reactive protein (CRP) levels and leucocyte counts unaffected. However, CiaHD increased lymphocyte counts ex vivo. Furthermore, we found a decrease in total CD3+CD4+CD69+ ((109/L), mean ± SD: acetate, 0.04 ± 1.0 versus citrate, 0.02 ± 0.01; P = 0.02) activated cells, while the number of CD28+ T cells remained stable. No differences were noted regarding T-cell exhaustion marker expression, CD14+CD16+ monocyte counts, and PMN-MDSCs. Compared with acetate, CiaHD has a minor impact on lymphocyte counts and CD4+T-cell activation, which was independent of systemic CRP and ionized magnesium, calcium levels, and other dialysis prescription modalities.

VIRAL CO-INFECTIONS IN PATIENTS WITH HIV-INFECTION

The paper presents the results of a study of the detection frequency of co-infectionserological markers of viral origin in persons suffering from HIV infection who are onantiretroviral therapy (ART). The relationship between the number of CD4 lymphocytesand the presence of markers of pathogens’ active replication of co-infections, caused byherpesviruses (cytomegalovirus – CMV, herpes simplex viruses of types 1 and 2 – HSV½,Epstein- Barr virus – EBV), and hepatitis B, C viruses in the plasma of HIV-infectedpatients, was considered.The purpose of the study – to determine the spectrum of viral co-infections in patientswith HIV infection and to find out the impact of these infections on the developmentof immunodeficiency based on the number of CD4 lymphocytes. Materials and methods. 146 patients with HIV infection, who were under outpatientobservation during 2020-2022 in the regional center of prevention and fight against AIDS inChernivtsi, were examined. When establishing the diagnosis, clinical and epidemiologicaldata and the results of laboratory tests, in particular: serological and immunological, weretaken into account. The level of CD4+ T-lymphocytes was examined after the disappearanceof the symptoms of the accompanying acute infectious disease (at least after 4 weeks).The results. The significance of the clinical manifestations and the nature of the course ofherpes simplex and shingles against a background of HIV infection depended on the degree ofreduction in the number of CD4+ lymphocytes. In most of the examined (57.0 %), the numberof CD4+ lymphocytes was less than 500 cells in 1 mm3 of blood, which contributed to theactivation of herpes simplex and shingles. A careful analysis of the frequency of detection ofserological markers of viral co-infections, depending on the route of HIV infection, showedthat hepatitis B virus markers were significantly more often detected in persons with a sexualroute of infection, and hepatitis C and mixed hepatitis virus markers – in those personsinfected with HIV, most likely, due to parenteral administration of narcotic substances.Conclusions. 1. Analysis of the spectrum of co-infections of viral genesis in HIV-infectedpersons demonstrated a wide distribution of parenteral viral hepatitis and a high levelof activation of chronic infections caused by HSV½, CMV and EBV. Markers of viralhepatitis B and C were detected in 56.8 % of patients with HIV infection. Activationof chronic infections, caused by CMV, HSV½ and EBV, was observed in 32.2 % of thesubjects. 2. Serological markers of viral hepatitis C and mixed hepatitis (HS + HBV)are most often detected in persons with parenteral HIV infection, and markers of viralhepatitis B – mostly in persons with sexual infection of HIV. 3. Antiretroviral therapyeffectively suppresses the replicative activity of the hepatitis B virus (HBV). 4. Theaverage values of the number of CD4-lymphocytes in HIV-infected persons who hadmarkers of co-infections of viral genesis, almost did not differ from the values of the levelof CD4-lymphocytes of those patients in whom these markers were not observed. Thistestifies to the effectiveness of antiretroviral therapy used in patients with HIV infection.

Flow Cytometric Analysis of Lymphocyte Subsets of Covid-19 Patients from A Single Centre in Turkey

Background:&#x0D; Lymphocyte counts have been shown to negatively correlate with the severity and different outcomes in Covid-19. The aim of this single centre study is to analyse the distribution of lymphocyte subsets in response to SARS-CoV-2 infection and its relation to the severity, course and prognosis of the disease.&#x0D; Methods:&#x0D; Blood samples were obtained from 67 consecutive patients between April 2020 and July 2020. Data on other laboratory parameters, and clinical course were collected retrospectively from patient files and patints were defined to have as severe or non-severe (mild/moderate) disease. Leukocyte subsets to be studied were identified by using flow cytometric analysis. Patients were allocated into 3 groups based on the day of blood sample collection for the flow cytometric analysis: Days 0-7, Days 8-14 and Days &gt;14 as Group I, Group II and Group III, respectively. In 10 available of 67 patients an additional flow cytometric analysis was done 7-10 days after the initial sampling.&#x0D; Results:&#x0D; Lower total lymphocyte, CD3 positive, CD4 positive and B-cell counts were identified in severe infection compared to non-severe infection group which were also correlated with high serum CRP, D-dimer and ferritin levels. NK and monocyte counts were not different between the two groups. Activation markers CD38 and HLA-DR on CD4 and CD8 positive lymphocytes also were not different in either group. &#x0D; Conclusion:&#x0D; CD3 and CD4 lymphopenia were lower in accordance with previous studies and were associated with severe disease. The expectancy of high activation markers was not met. Future studies with detailed subgroup analyses at different time-points, and immune profiling after vaccination as well as during new infection in vaccinated patients will shed more light on our general knowledge of the immune response to COVID-19 and viruses in general.