Number Of CD34 Cells Research Articles

Allowable total error in CD34 cell analysis by flow cytometry based on state of the art using Spanish EQAS data.

CD34+ hematopoietic stem cell (HSC) enumeration, crucial for HSC transplantation, is performed by flowcytometry to guide clinical decisions. Variability in enumeration arises from biological factors, assay components, and technology. External quality assurance schemes (EQAS) train participants to minimize inter-laboratory variations. The goal is to estimate total error (TE) values for CD34 cell enumeration using state-of-the-art (SOTA) methods with EQA data and to define quality specifications by comparing TE using different cutoffs. A total of 3,994 results from 40 laboratories were collected over 11 years (2011-2022) as part of the IC-2 Stem Cells Scheme of the GECLID Program that includes absolute numbers of CD34 cells. The data were analyzed in two periods: 2011-2016 and 2017-2022. The TE value achieved by at least 60 %, 70 %, 80 %, and 90 % of laboratories was calculated across the two different periods and at various levels of CD34 cell counts: above 25, 25 to 15, and under 15 cells/μL. A decrease in the SOTA-based TE for CD34 cell enumeration was observed in the most recent period in 2017-2021 compared with 2012-2016. A significant increase of P75 TE values in the low CD34 range (<15 cells/μL) levels was found (p<0.001). Technical advancements contribute to the decrease TE over time. The TE of CD34 cell FC counts is measure-dependent, making it responsive to precision enhancement strategies. The TE measured by EQAS in this study may serve as a quality specification for implementing ISO 15189 standards in clinical laboratories for CD34 cell enumeration.

HIV tropism to chemokine coreceptors. Features of the definition, the current state

HIV infection is a significant cause of death worldwide, the number of people with HIV infection in the Russian Federation as of 2022 amounted to 1163818 people. The determination of HIV tropism is necessary for the appointment of drugs from the group of penetration inhibitors, and also opens up new opportunities in the prediction and analysis of HIV infection in a patient.Aim. To present a description and assess the current state of methods for determining HIV tropism, to summarize known information about the influence of HIV tropism on the course of the disease, to identify topical issues related to HIV tropism and requiring solutions.Materials and methods. A review of domestic and foreign sources devoted to methods for determining the prevalence and clinical significance of HIV tropism was carried out.Results. For the effective administration of CCR5 antagonist drugs, preliminary analysis is necessary to establish the tropism of HIV by genotypic or phenotypic methods. The use of CCR5 antagonists is not possible if HIV can use the CXCR4 coreceptor. CXCR4 – tropism of HIV is associated with the duration of the disease, a decrease in the number of CD4 cells, AIDS, and is a negative prognostic factor. Human mutations affecting coreceptors can affect the course of infection and susceptibility to HIV.Conclusion. The determination of HIV tropism is a useful analysis, the importance of which will increase in connection with the development of new drugs from the group of penetration inhibitors. To increase the availability of HIV tropism analysis in the Russian Federation, the creation of genotypic test systems is required. To create proprietary algorithms used in genotypic analysis, as well as laboratory testing and development of new effective drugs from the group of penetration inhibitors, it is necessary to develop a phenotypic test system. The small study of the influence of other regions of the env gene on HIV tropism, the study of HIV tropism to alternative coreceptors are urgent issues that need to be addressed.

P53 ABNORMAL ENDOMETRIAL CANCERS – ANALYSIS OF PD-1 CO-EXPRESSION BY CD4 HELPER AND CD8 CYTOTOXIC T-CELL SUBTYPES IN THE TUMOR MICROENVIRONMENT

Abstract Introduction/Objective Endometrial cancer is classified into four molecular subtypes of which the p53 abnormal variant is biologically aggressive. Programmed cell death protein 1 (PD-1) is a cell surface receptor and a vital inhibitor of the immune response. Programmed death ligand 1 (PD-L1) is a trans-membrane co-inhibitory factor of the immune response. PD-1 / PDL-1 interaction inhibits the activation and survival of T cells and decreases cytokine secretion. Some cancer cells express PD-L1, which then bind to PD-1 expressed on tumor specific T cells, and escape destruction by the T cells. The CD4/CD8 T cell immune response in the p53 abnormal variant has not been adequately evaluated with respect to the PD-1/PD-L1 pathway. Understanding this immune microenvironment is important to assess the potential use of anti-PD-1 targeted tumor therapy. Methods/Case Report Twenty cases of p53 abnormal endometrial cancers were obtained. All cases were PDL-1 +ve by immunohistochemistry. Two dual stains were performed - PD-1/CD4 and PD-1/CD8 antibodies. Six high-power fields (40x objective) with the greatest concentration of lymphocytes were counted. The total number of CD4 or CD8-only cells was recorded for each of the two stained slides for each case. The total number of PD- 1/CD4 and PD-1/CD8 positive cells were counted and recorded. The results were tabulated, and a paired t-test statistical analysis was performed to compare the total number of CD4 and CD8 cells and the percentage of PD-1/CD4 cells with the percentage of PD-1/CD8 positive cells. Results (if a Case Study enter NA) A statistically significant increase in the number of CD8 (Mean = 383) compared to CD4 cells (Mean = 317) was seen in the tumor microenvironment (p-value=0.0023). Evaluating PD-1 expression, the converse was true, and the percentage of PD-1/CD4 cells (Mean = 13.035%) was increased compared to the percentage of PD-1/CD8 cells (Mean = 9.205%), achieving statistical significance (p-value=0.0116). Conclusion The statistically significant increase in CD8 cells in the tumor and its stroma suggests that a robust cytotoxic response is being mounted to this high-risk molecular variant. The concurrent increase in CD4/PD-1 positive lymphocytes provides an appropriate microenvironment for the use of anti-PD-1 immunotherapy, since blockade of PD-1 on CD4 helper T cells may augment the tumor ablative cytotoxic properties of the CD8 positive lymphocytes. This includes reinvigoration of exhausted CD8+ T cells. Anti-PD-1 may also prevent the inhibition of PD-1 positive CD8 cells.

Two 5-year studies of bictegravir/emtricitabine/tenofovir alafenamide in people with HIV: a plain-language summary.

This is a plain-language summary of an article that reported on two studies of the medication bictegravir/emtricitabine/tenofovir alafenamide (shortened to B/F/TAF). B/F/TAF is a single pill containing three different drugs used to treat human immunodeficiency virus (known as HIV). The drugs work together to lower the levels of HIV (called viral load) in the body and make the virus undetectable in the blood. Researchers measured whether B/F/TAF was safe and effective when taken over 5years in over 400 people in 10 countries who had never taken HIV medication before. After 5years, almost all (99%) of the people who took B/F/TAF had an undetectable viral load. This does not mean that they were cured, but that the levels of HIV were so low that the tests used by researchers could not detect the virus in the blood. CD4 is a type of immune system cell. HIV causes CD4 cell numbers to decrease. On average, the number of CD4 cells increased by more than 300 cells per microliter (cells/μL) of blood over 5years. This means that the immune system was improving. HIV is able to change its genes to escape the effects of the drugs. This is known as HIV resistance to treatment. Nine people had a viral load high enough to suggest that the drugs might not be working, but no resistance to B/F/TAF was seen. Some people (less than one in three) experienced medical problems thought to be linked to B/F/TAF treatment, known as side effects. The most common side effects were headache, diarrhea, nausea, tiredness (fatigue), dizziness, and difficulty falling or staying asleep (insomnia). On average, people's body weight increased by 3kg in the first year of taking B/F/TAF. This might be because their general health improved after starting HIV treatment. Weight gained after that time was similar to the level of weight gain expected in the general population. Very few people (less than 1 in 100) stopped taking B/F/TAF because of side effects thought to have been caused by B/F/TAF. B/F/TAF was effective at treating HIV in people who had never taken HIV medication before. Most (70%) people were still taking B/F/TAF after 5years.Clinical Trial Registration: NCT02607930 (Study 1489); NCT02607956 (Study 1490) (ClinicalTrials.gov).

Burn Wound Healing Activity of Hydroxyethylcellulose Gels with Different Water Extracts Obtained from Various Medicinal Plants in Pseudomonas aeruginosa-Infected Rabbits.

Burn injuries represent a significant problem in clinical practice due to the high risk of infection and the prolonged healing process. Recently, more attention has been given to natural remedies such as water extracts of various medicinal plants, which possess anti-inflammatory and wound healing properties. The aim of this study is to evaluate the efficacy and safety of Satureja montana L. and other water extracts in a burn wound model. The study involved male Californian rabbits (n = 52) divided into eight groups. Burn wounds were modeled on the animals and subsequently treated with gels based on Satureja montana L. and other water extracts. The reparative potential of the epidermis (assessed by Ki-67 expression), the state of local immunity (measured by the number of CD-45 cells), and the anti-inflammatory role of mast cells (measured by tryptase levels) were evaluated. Bacteriological and morphological studies were conducted. The most pronounced bactericidal, reparative, and immunostimulatory effects were observed after the treatment using a gel mixture of water extracts from Satureja montana L., Salvia sclarea, Coriandrum sativum L., and Lavandula angustifolia in equal proportions (1:1:1:1). The other gels also demonstrated high efficacy in treating burn wounds, especially when using a strain of Pseudomonas aeruginosa resistant to several antibiotics. Immunohistochemical studies showed a significant increase in the number of Ki-67-positive cells in the basal layer of the epidermis and a decrease in the number of CD-45-positive cells, indicating improved proliferative activity and reduced inflammation. This study confirms the hypothesis that the use of water extract mixtures significantly enhances the reparative potential, improves the immune response in the treatment of burns, and promotes wound healing. These findings pave the way for further research and the application of complex phytotherapeutic agents, specifically water extracts of medicinal plants containing phenols and antioxidants in burn wound therapy.

Морфофункциональное состояние органов иммунной системы у поросят-гипотрофиков в ранний неонатальный период

Abstract. The purpose of the study was to study the state of the immune system organs in hypotrophic piglets in the early neonatal period in an industrial pig breeding complex. Methods. The experiment was conducted in 2023 in a large industrial pig farm in the Voronezh region on piglets of the wounded neonatal period received from sows 3-4 farrowing. The sows were kept at optimal microclimate parameters, taking into account their physiological state, and fed with SK-1 nutritionally balanced feed. At the initial stage of the experiment, the piglets obtained during farrowing underwent clinical examination and weighing. Animals under 800 g are counted as hypotrophic piglets (n = 30), animals over 800 g are normotrophic, respectively (n =30). After the groups were formed, animals were forced to be slaughtered before taking colostrum (n =5) and biological material was taken from each group (thymus, lymph nodes (inguinal), spleen for immunohistochemical studies. Results. The level of mitotic activity in the thymus of normotrophic piglets was 9.4 % higher (p &lt; 0.05) than in piglets with body weight deficiency. In the spleen, the level of mitotic activity had no significant differences and was approximately the same in all animals participating in the experiment. In lymph nodes, the mitotic activity of cells in normotrophics was 12.7 % higher (p &lt; 0.05). A study of positively expressed CD-3 cells in the spleen revealed significant differences, so, in normotrophic piglets, the number of cells positively stained with this marker was significantly higher than in piglets with a body weight deficiency by 10.2 % (p &lt; 0.05). The number of CD-3 cells in the thymus differed by 5.6 %, and in the lymph nodes by 2.4 % between the groups, but there were no significant differences. The cellular expression of “immature” forms of B lymphocytes (PAX-5) in the lymph nodes of normotrophics was significantly higher by 12.9 % (p &lt; 0.05) compared with hypotrophic piglets. The scientific novelty lies in the fact that for the first time a comprehensive immunohistochemical study of the organs of the immune system of piglets in the early neonatal period was carried out using monoclanal antibodies CD-3, Ki-67 and PAX-5. As a result of the experiment, it was revealed that piglets with body weight deficiency have a “depression” of the immune system, which manifests itself in hypoplasia of T-lymphocytes in the spleen, “maturing” B-lymphocytes in the lymph nodes and low mitotic potential in the thymus and lymph nodes.

Dynamics of Matricellular Protein Levels in Blood Predict Recovery in Patients with Human Immunodeficiency Virus-Tuberculosis Coinfection.

Chronic immune activation in tuberculosis (TB) associated with human immunodeficiency virus (HIV) infection (HIV/TB) modifies their clinical course. We prospectively measured osteopontin (OPN), full-length galectin-9 (FL-Gal9), and total-Gal9 (T-Gal9) levels in 32 patients with HIV/TB coinfection treated with anti-tuberculosis and antiretroviral therapies over 6-18 months to determine the amelioration of inflammatory conditions in response to the therapies. We observed a significant time-dependent decrease in FL-Gal9 in both pulmonary TB (PTB, n = 20) and extrapulmonary TB (EPTB, n = 12) patients. The levels of T-Gal9, OPN, and CRP decreased significantly after treatment in only PTB patients. We calculated the inflammatory score (INS) indicating immunologic recovery based on the decline in OPN, FL-Gal9, T-Gal9, and CRP levels. Baseline levels of T-Gal9 and OPN positively correlated with INS in all TB and only PTB patients, respectively, indicating that their levels predict better recovery. In contrast, FL-Gal9 levels at the second visit negatively correlated with INS in EPTB patients. The decrease rate in OPN levels at the second visit also correlated positively with INS in PTB patients. Women showed a higher INS and lower levels of FL-Gal9 than men. The patients with moderate grade severity on chest X-ray had higher CD4 cell numbers than those with limited grade severity. Monitoring these markers will help to predict and assess the response to therapy as well as to devise strategies to reduce the complications caused by chronic immune activation in patients with HIV/TB coinfection.

Immunohistochemical characterization of the immune cell response during chlamydial infection in the male and female koala (Phascolarctos cinereus) reproductive tract.

Chlamydiosis is one of the main causes of the progressive decline of koala populations in eastern Australia. While histologic, immunologic, and molecular studies have provided insights into the basic function of the koala immune system, the in situ immune cell signatures during chlamydial infection of the reproductive tract in koalas have not been investigated. Thirty-two female koalas and 47 males presented to wildlife hospitals with clinical signs suggestive of Chlamydia infection were euthanized with the entire reproductive tract collected for histology; immunohistochemistry (IHC) for T-cell (CD3ε, CD4, and CD8α), B-cell (CD79b), and human leukocyte antigen (HLA)-DR markers; and quantitative real-time polymerase chain reaction (rtPCR) for Chlamydia pecorum. T-cells, B-cells, and HLA-DR-positive cells were observed in both the lower and upper reproductive tracts of male and female koalas with a statistically significant associations between the degree of the inflammatory reaction; the number of CD3, CD4, CD79b, and HLA-DR positive cells; and the PCR load. CD4-positive cells were negatively associated with the severity of the gross lesions. The distribution of immune cells was also variable according to the location within the genital tract in both male and female koalas. These preliminary results represent a step forward towards further exploring mechanisms behind chlamydial infection immunopathogenesis, thus providing valuable information about the immune response and infectious diseases in free-ranging koalas.

LncRNA XIST Interacts with Regulatory T Cells within the Tumor Microenvironment in Chronic Hepatitis B-Associated Hepatocellular Carcinoma.

Alterations in the expression of several long non-coding RNAs (lncRNAs) have been shown in chronic hepatitis B-associated hepatocellular carcinoma (CHB-HCC). Here, we aimed to investigate the association between the expression of inflammation-associated lncRNA X-inactive specific transcript (XIST) and the type of inflammatory cells within the tumor microenvironment. Twenty-one consecutive cirrhotic patients with CHB-HCC were included. XIST expression levels were investigated on formalin-fixed paraffin-embedded (FFPE) tumoral and peritumoral tissue samples by real-time polymerase chain reaction (RT-PCR). Immunohistochemical staining for CD3, CD4, CD8, CD25, CD163, CTLA4, and PD-1 were performed. The findings were statistically analyzed. Of the 21 cases, 11 (52.4%) had tumoral and 10 (47.6%) had peritumoral XIST expression. No significant association was found between the degree of inflammation and XIST expression. The number of intratumoral CD3, CD4, CD8 and CD20 positive cells was higher in XIST-expressing tumors, albeit without statistical significance. Tumoral and peritumoral XIST expression tended to be more common in patients with tumoral and peritumoral CD4high inflammation. The number of intratumoral CD25 positive cells was significantly higher in XIST-expressing tumors (p=0.01). Tumoral XIST expression was significantly more common in intratumoral CD25high cases (p=0.04). Peritumoral XIST expression was also more common among patients with CD25high peritumoral inflammation, albeit without statistical significance (p=0.19). lncRNA XIST is expressed in CHB-HCC and its expression is significantly associated with the inflammatory tumor microenvironment, particularly with the presence and number of CD25 (+) regulatory T cells. In vitro studies are needed to explore the detailed mechanism.

Senescent Immune Signature in Frail and Vulnerable Newly Diagnosed Myeloma Patients: A Cohort Analysis from the UK-MRA Myeloma XIV Fitness Study

Introduction Physical frailty is a key factor driving treatment tolerability and outcome in multiple myeloma (MM). There is a close interplay between physiological aging, frailty and immune dysfunction, with the presence of a malignant disease overlay adding an additional dimension to the host response biology. The UKMRA Myeloma XIV FiTNEss study is the first phase 3 randomised clinical trial to explore the utilisation of frailty assessment as a tool to guide and optimise treatment decision making and deliverability. Here we report on the relationship between physical frailty and T cell dysfunction in a cohort of patients within this landmark study. Methods Multiparameter flow cytometry was used to assess the baseline T cell phenotype in the peripheral blood of 49 participants with NDMM receiving treatment within FiTNEss. Patients were assessed for frailty using the IMWG frailty score and additionally assessed for clinical vulnerability using the UKMRA MRP score (Cook et al, Lancet Haematol. 2019). The IMWG frailty score combines activities of daily living assessment and co-morbidity burden with age to generate a validated, prognostic frailty score. The UKMRA MRP is a risk profile which uses routinely collected clinical data including age, WHO performance status, ISS stage and CRP to define prognostic risk categories with the high-risk category encompassing frailer individuals. Normal T cell subsets were assessed based on CD45RA and CD28 phenotype while dysfunctional subsets (exhausted and senescent) were defined using CD28 and KLRG1. Results The median age of participants was 79 (range 68-90) and 55% were male. The distribution by IMWG was fit 18%, unfit 33%, frail 49% and by MRP was low risk 31%, intermediate risk 28%, high risk 38%. Absolute lymphocyte count (ALC) decreased with increasing frailty (p=0.02), with no patients classified as fit by IMWG or low risk by MRP criteria having a lymphocyte count below the lower limit of normal. No relationship was seen between ALC and age. The ratio remained stable across the IMWG cohorts, suggesting a global reduction in lymphocyte subsets. An accumulation of CD8 TEMRA cells was seen in the MRP high risk cohort (p=0.0085). No differences were seen between cohorts based on the IMWG frailty score. The accumulation of CD8 TEMRA cells indicates a loss of T cell differentiation plasticity and is associated with a cytotoxic and pro-inflammatory immune response. CD4 dysfunctional subsets were comparable across the cohorts. CD8 dysfunctional subsets were most marked in the frail and high-risk cohorts. The MRP high-risk cohort had a skewed ratio of exhausted to senescent cells (p=0.02), with a reduction in CD8 exhausted populations compared to the fit and unfit cohorts (p=0.0477) and a trend towards increased senescent cell burden (p=0.07). This was echoed within the IMWG frail cohort where elevated numbers of senescent CD8 cells were identified (p=0.02). No relationship was seen between CD8 senescent burden and age, CRP, ISS or WHO PS when each variable was assessed independently, suggesting that the presence of this immune population is not directly related to disease burden or physiological aging alone, but reflects a more complex interplay between host response biology and disease characteristics. Principal component analysis demonstrated a high degree of immunological heterogeneity within the MRP low and intermediate risks cohorts, while the high-risk cohort had a more uniform immunological profile (Figure 1A). Significant overlap between a proportion of the non-high-risk cohorts and high-risk cohorts was seen and requires further exploration. A multi-variable correlation analysis identified correlations between the CD4:8 ratio and CD8 cell senescent burden, MRP and ISS, highlighting the important interplay between immunological dysfunction, clinical vulnerability and disease burden (Figure 1B). Conclusion The T cell immunological landscape of the most vulnerable patient populations with newly diagnosed MM is characterised by an accumulation of senescent CD8 T cells which cannot be attributed to age or disease burden alone. Immune senescence is known to be associated with a chronic inflammatory state which may drive the acquisition of the physiological changes, such as sarcopenia, which characterise a frailty phenotype. This provides an opportunity for targeted intervention to address the biological drivers of disease related frailty.

Melatonin inhibits the formation of chemically induced experimental encapsulating peritoneal sclerosis through modulation of T cell differentiation by suppressing of NF-κB activation in dendritic cells

Encapsulating peritoneal sclerosis (EPS) is a severe complication of peritoneal dialysis (PD). Surgery is a therapeutic strategy for the treatment of complete intestinal obstruction. However, complete intestinal obstruction in long-term PD results in high mortality and morbidity rates after surgery. Immunopathogenesis participates in EPS formation: CD8, Th1, and Th17 cell numbers increased during the formation of EPS. The anti-inflammatory and immunomodulatory effects of melatonin may have beneficial effects on this EPS. In the present study, we determined that melatonin treatment significantly decreases the Th1 and Th17 cell populations in mice with EPS, decreases the production of IL-1β, TNF-α, IL-6, and IFN-γ, and increases the production of IL-10. The suppression of Th1 and Th17 cell differentiation by melatonin occurs through the inhibition of dendritic cell (DC) activation by affecting the initiation of the NF-κB signaling pathway in DCs. Our study suggests that melatonin has preventive potential against the formation of EPS in patients with PD.

#6158 UREMIC TOXINS ARE ASSOCIATED WITH IMMUNE-SENESCENCE AND IMMUNE-EXHAUSTION IN HEMODIALYSIS PATIENTS

Abstract Background and Aims Immune-senescence and exhaustion have been described in hemodialysis (HD) patients, with changes in lymphocyte phenotype possibly stimulated by the uremic environment, independent of age and comorbid conditions. We evaluated the potential association of uremic toxin accumulation with changes in lymphocyte count and phenotype in HD patients. Method In the peripheral blood of 54 HD patients (23 female, mean age 51.3±16.91 years) and 31 age-matched healthy controls, lymphocytes surface molecules were determined by flow cytometry: T cells (CD4, CD8, CD31, CD45RA, CCR7, CD28, CD57, PD1) and B cells (CD27, IgD). Plasma levels of the protein bound uremic toxins [hippuric acid (HA), indoxyl sulfate (IxS) and p-cresyl sulfate (pCS)] were measured by ultra-performance liquid chromatography. Lymphocyte phenotypes were correlated with uremic toxins levels using Spearman's correlation coefficient test. Patients with recent or active infection, malignancy, autoimmune disease and/or diabetes mellitus were excluded. Results A higher concentration of uremic toxins was observed in HD patients, compared to healthy controls, namely: total HA 3.05 (1.66-5.37) vs 0.102 (0.04-0.2)mg/dl (p&amp;lt;0.001), free HA 1.482 (0.7-2.8) vs 0.029 (0.03-0.04)mg/dl (p&amp;lt;0.001); total IxS 2.207 (1.27-3.34) vs 0.063 (0.04-0.09)mg/dl (p&amp;lt;0.001), free IxS 0.146 (0.09-0.27) vs 0.0004 (0.0004-0.0004)mg/dl (p&amp;lt;0.001); total pCS 1.248 (0.84-1.66) vs 0.066 (0.04-0.13)mg/dl (p&amp;lt;0.001), free pCS 0.089 (0.06-0.13) vs 0.004 (0.004-0.005)mg/dl (p&amp;lt;0.001); respectively. There was a positive correlation between hemodialysis vintage and total as well as free HA (p = 0.005, p = 0.003, respectively). No significant differences in uremic toxin levels were found in HD patients according to sex, age, dialysis prescription and method. Patients with residual urine output had reduced total and free HA levels 1.739 (3.5) vs 3.347 (4.16)mg/dl (p = 0.03), and 0.717 (1.9) vs 1.901 (3.5)mg/dl (p = 0.04), respectively and free IxS levels, 0.117 (0.1) vs 0.177 (0.2)mg/dl (p = 0.02), respectively. Total lymphocyte count and absolute number of CD4 cells were negatively correlated with total (p = 0.018 and p = 0.02 respectively) and free HA levels (p = 0.024 and p = 0.017 respectively). Several naïve and less differentiated CD4 subpopulations also demonstrated a negative correlation with total and free HA levels, most importantly CD4CD31+ (p = 0.037 and p = 0.027 respectively), CD8CD28+CD57- (p = 0.018 and p = 0,014 respectively) CD4CD45+CD57- (p = 0.039 and p = 0.027 respectively) and naïve B cells (CD19+IgD+CD27-) (p = 0.042 and p = 0.032 respectively). Advanced differentiated lymphocyte subtypes, CD4CD28+CD57+ and CD4CD45-CD57+ had a positive correlation with free ΗΑ levels (p = 0.036 and p = 0.045 respectively). Exhausted CD4 cells, defined as CD4+PD1+ had positive correlations with total and free IxS levels (p = 0.004 and p = 0.011 respectively) and total and free pCS (p = 0.027 and p = 0.018 respectively), but not with HA levels. Moreover, further divided exhausted lymphocytes, according to CD45RA expression, CD4CD45RA-PD1+ cells correlated positively with total and free IxS levels (p = 0.004 and p = 0.01 respectively) and CD4CD45RA+PD1+ with pCS total and free levels (p = 0.039 and p = 0.045 respectively). Conclusion Uremic toxins are increased in hemodialysis patients and may influence adaptive immunity. HA is associated with an immune-senescent, while IxS and pCS to an immune-exhausted lymphocyte phenotype.

High density and proximity of CD8+ T cells to tumor cells are correlated with better response to nivolumab treatment in metastatic pleural mesothelioma.

The efficacy of immune checkpoint inhibitors (ICIs) in pleural mesothelioma has recently been established. The response to ICIs can be predicted by quantitative analysis of cells and their spatial distribution in the tumor microenvironment (TME). However, the detailed composition of the TME in pleural mesothelioma has not been reported. We evaluated the association between the TME and response to ICIs in this cancer. A retrospective analysis of 22 pleural mesothelioma patients treated with nivolumab in different centers was performed using surgical specimens. Four patients had a partial response to nivolumab (response group) and 18 patients had stable or progressive disease (nonresponse group). The number of CD4, CD8, FoxP3, CK, and PD-L1 positive cells, cell density, and cell-to-cell distance were analyzed by multiplex immunofluorescence. PD-L1 expression did not differ significantly between the response and nonresponse groups. The density of total T cells and of CD8+ T cells was significantly higher in the response than in the nonresponse group. CD8+ T cells were more clustered and located closer to tumor cells, whereas regulatory T cells were located further from tumor cells in the response than in the nonresponse group. High density and spatial proximity of CD8+ T cells to tumor cells were associated with better response to nivolumab, whereas the proximity of regulatory T cells to tumor cells was associated with worse response, suggesting that the distinct landscape of the TME could be a potential predictor of ICI efficacy in pleural mesothelioma.

Does COVID-19 Vaccination with BNT162b2 Influence HIV-Related Immunological and Virological Markers? Data from 235 Persons Living with HIV at Cotugno Hospital, Naples, Italy: Immune Response After Second and Third Doses, and Influence on Immunovirological Markers.

Few data are available on the impact of COVID-19 vaccination on CD4 counts and HIV-RNA in persons living with HIV (PLWH). We present the data of 235 PLWH who were vaccinated with BNT162b2 in March 2021-February 2022 at the "Cotugno" hospital in Naples. PLWH treated at the "Cotugno" hospital, who were vaccinated at the hospital vaccination center, without prior COVID-19 and for whom immunological/virological data were available in the last 12 months and in the 6 months after vaccination were included. Antispike Ab were available for 187 and 64 PLWH after the second and third doses: PLWH with antispikes >33 binding antibodies units (BAU)/mL increased from 91% to 98%. Antinucleocapsid Ab performed in 147 and 56 patients identified 19 (13%) asymptomatic/paucisymptomatic COVID-19 infections after the second dose and an additional 15 (27%) after the third dose. Immunological/virological data were collected before vaccination (T0), after the second dose (T1), and after the third dose (T2). The absolute number of CD4 increased after the third dose (median 663, 657, and 707 at T0, T1, and T2; p < 0.000 T0 vs. T2). The proportion of patients with HIV-RNA <50 copies/mL increases significantly after the second dose (73%; 85.7%; 87.7%; p < 0.000 T0 vs. T2). The presence of COVID-19 asymptomatic/paucisymptomatic infections (demonstrated by the presence of antinucleocapsid Ab) significantly increases SARS-CoV-2 antispike Ab after second dose, but not after third dose. Asymptomatic/paucisymptomatic COVID-19 infections do not have influence on CD4 cell number and HIV-RNA level. Similarly, the presence of not-controlled HIV-RNA (HIV-RNA >50 copies/mL) does not influence antispike Ab response. According to our data, the response to SARS-CoV2 vaccination is effective in people living with HIV. Vaccination against COVID-19 appears to positively affect immunological and virological levels in people living with HIV.

Evaluation of CD4 Count, Haemoglobin, and Intestinal Parasites Status of Cancer Patients Receiving Oncology Therapy in Calabar, Nigeria

Cancer is a human cellular error with a high mortality rate, especially in developing regions. Resolving this error with therapy may render patients immunologically depressed and susceptible to opportunistic infections including parasitic. The gut parasites, Cluster of Differentiation four (CD4) cells and haemoglobin (Hb) levels of cancer patients on oncology therapy in Calabar, Nigeria were studied. Stool and blood samples were collected from 317 (186 cancer and 131 apparently healthy) subjects. Parasites were identified using direct smear and formol ether concentration techniques while CD4 and Hb estimation were done using BD-fascount and fluorescent flow cytometric techniques, respectively. Data were analysed using chi-square and independent sample t-test. Overall parasitosis prevalence in this study was (27.44%) with cancer showing an association with intestinal parasitosis (P=0.001) and CD4 level (P=0.002), respectively but none with Hb level. Parasiteinfected cancer subjects had significantly lower mean haemoglobin concentration than their non-infected counterpart (t = - 4.869, P&lt;0.001). Generally, cancer subjects on cancer treatment had an insignificantly lower parasite and significantly lower CD4 levels compared to their non-treated counterpart (P = 0.829) and (P&lt;0.001), respectively but their mean haemoglobin levels were similar (P=0.701). Cancer subjects on chemotherapy alone had significantly higher parasite prevalence, 50.00% compared to 28.95% for those on combined (surgery and chemotherapy) treatment (P=0.014), whereas the type of treatment showed no significant effect on CD4 and Hb levels. Parasite occurrence was only pronounced (71.43%) among subjects with &gt;12 months of treatment duration (P=0.1038). There was a correlation between duration of treatment and CD4 levels (r = - 0.231, P&lt;0.05) but none with haemoglobin levels (r = 0.024, P&gt;0.05) of cancer subjects. In conclusion, although this study has shown that cancer as a condition increases parasite prevalence and reduces mean CD4 counts in subjects, their haemoglobin levels were not significantly affected, except for subjects with intestinal parasite infections. Treatment with cancer therapy did not also affect the haemoglobin concentration, but significantly reduced the CD4 cell number of subjects. Subjects who had majorly, chemotherapy were mostly infected with the parasite but with no noticeable effect on their CD4 and haemoglobin levels. A prolonged duration of treatment increased parasitosis and reduced the CD4 level of subjects but did not affect haemoglobin levels. Apart from the oncological treatments given to cancer subjects, management of cancer subjects should include parasitological examination and treatment, periodic monitoring of their hematologic profile and regular immune revitalization procedures, especially for those placed on prolonged therapy.

Effect of characteristics on the clinical course at the initiation of treatment for human immunodeficiency virus infection using dimensionality reduction

The beginning of human immunodeficiency virus (HIV) infection treatment depends on various factors, which are significantly correlated with the initial CD4 cell number. However, a covariate correlation between these factors may not reflect the correct outcome variable. Thus, we evaluated the effects of a combination of fixed factors (reduced dimensions), which determine when to start treatment for the first time, on short-term outcome, long-term outcome, and survival, considering correlations between factors. Multiple correspondence analysis was performed on variables obtained from 925 patients who participated in a Korean HIV/acquired immunodeficiency syndrome cohort study (2006–2017). Five reduced dimension groups were derived according to clinical data, viral load, CD4 cell count at diagnosis, initial antiretroviral therapy, and others. The dimension group with high initial viral loads (55,000 copies/mL) and low CD4 cell counts (< 200 cells/mm3) should start treatment promptly after diagnosis. Groups with high initial CD4 cell counts (> 350 cells/mm3) that did not require immediate treatment according to previous guidelines had a higher failure rate for long-term relative CD4 recovery. Our results highlight the importance of early diagnosis and treatment to positively influence long-term disease outcomes, even if the initial immune status is poor, given the patient’s combination of early diagnostic symptoms.

Mobilization of Autologous Peripheral Blood Stem Cells (PBSC) in CD20+ Lymphoma Patients Using RICE, Granulocyte-Colony Stimulating Factor (G-CSF) and Plerixafor

RICE is salvage therapy for treating CD20+non-Hodgkin lymphoma (NHL). It is combined with G-CSF to collect autologous peripheral blood stem cells (aPBSC). Little data exists, though, on the combination of G-CSF and Plerixafor after RICE in mobilizing adequate number of CD34 cells and the product’s immune content. We report on the results of twenty CD20+ NHL patients after RICE, G-CSF and Plerixafor were given to collect aPBSC. The median number of cells collected was 12.92 × 10&lt;sup&gt;6&lt;/sup&gt; CD34 cells/kg (range 5.44-83.76). Plerixafor toxicity included diarrhea (n = 5) and injection-site irritation (n = 1). Seventeen patients collected; fifteen patients did so in one session. Two patients with CLL/Richter or transformed follicular had positive-flow products. Addition of Plerixafor to G-CSF increased by 2.6-9 folds the number of blood CD34 cells. Sixteen patients went to ASCT, with a median of 7.29 × 10&lt;sup&gt;6&lt;/sup&gt; CD34 cells/kg infused. The median engraftment time post-ASCT for neutrophils was 12 (range 10-19), for platelets ≥20K 11 (range 0-19) and ≥50K 16.5 (range 11-42) days. There were no graft failures. In APBSC product, there was no evidence of NK or LAK lytic activity (n = 10), only LAK activity (n = 4) and both LAK and NK activity (n = 2). Blood NK activity was common on day +28 post-ASCT. There was no significant correlation between apheresis product and the number of blood immune cells post- ASCT or relapse. Addition of Plerixafor to RICE/G-CSF is well tolerated. The majority of patients collected aPBSC in one session.

Copper Repletion Restores the Number and Function of CD4 Cells in Copper-Deficient Rats

Dietary copper deficiency decreases the number of splenic CD4 cells and mitogen-induced generation of interleukin-2 activity and DNA synthesis in cultures of splenic mononuclear cells. To determine the reversibility of these defects, Cu-deficient rats were fed a Cu-adequate diet for either 4, 7 or 11 d before preparation of cell cultures. Serum and hepatic concentrations of Cu attained 87 and 75%, respectively, of the control level after 4 d of dietary repletion. In contrast, interleukin-2 activity and [3H]thymidine incorporation in splenic cell cultures treated with T-cell mitogens were significantly greater than in cultures from Cu-deficient rats after 7, but not 4, d of dietary Cu repletion. The number of splenic CD4 cells was also greater after 7 d of dietary supplementation with Cu. Changes in the relative percentage and function of T-helper cells were highly correlated with one another and with hepatic Cu concentration. These observations indicate that an inadequate supply of dietary Cu reversibly suppresses the maturation and function of splenic T-helper cells.

Herbal Honey Preparations of Curcuma Xanthorriza and Black Cumin Protect against Carcinogenesis through Antioxidant and Immunomodulatory Activities in Sprague Dawley (SD) Rats Induced with Dimethylbenz(a)anthracene

Traditionally, Curcuma xanthorriza (CX), black cumin seed (BC), and honey have been used by the Indonesian people as medicinal ingredients to treat various health symptoms. CX extracts and BC have been proven in the laboratory as chemopreventive agents, antioxidants, and immunomodulators. In this study, we developed CX extract, BC oil, and honey into herbal honey preparations (CXBCH) and hypothesized that the preparations show chemopreventive activity. The purpose of the study was to determine the CXBCH potential as chemopreventive, antioxidant, and immunomodulatory. In this experimental laboratory research, antioxidant, immunomodulatory, and cytotoxic activities were tested on human mammary cancer cell lines (T47D cells) while the chemopreventive activity of the CXBCH preparations on Sprague Dawley (SD) rats induced with dimethylbenzene(a)anthracene (DMBA). CXBCH preparations demonstrated immunomodulatory, antioxidant, and cytotoxic activities in T47D, Hela, and HTB-183 cells and in DMBA-induced SD rats, as the preparations inhibited tumor nodule formation, increased the number of CD4, CD8 and CD4CD25 cells, and glutathione-S-transferase (GST) activity, and decreased serum NO levels. CXBCH preparations display chemopreventive, antioxidant, and immunomodulatory properties.

Discordant immune response among treatment experienced patients infected with HIV-1: Crosstalk between MiRNAs expression and CD4+ T cells count.

One of the problems with treating HIV-infected patients with ARVs is that the treatment can reduce viral load and does not increase the number of CD4 cells (immunological discordance). There are still challenges to treating HIV-positive patients. This study aimed to investigate the expression level of 18 miRNAs involved in the proliferation and differentiation of CD4+ T cells in a target (discordant immune response) and a control (immune response) group. In this case-control study, 18 miRNAs were selected and synthesized according to the in-silico analysis and published literatures. RNA extraction was performed from PBMC cells of 30 HIV-1 positive patients in the sample bank. The expression level of microRNAs was calculated by the relative q PCR method (2-ΔΔCt method), and data were analyzed using GraphPad Prism software version 8.0.2. The results of fold change calculation and statistical analysis showed that the expression levels of miR-30b (p value: 0.01, fold change: 0.23), miR-155 (p value: 0.04, fold change: 0.44), miR-181a (p value: 0.01, fold change: 0.37), and miR-190b (p value: 0.01, fold change: 0.39) had a significant decrease in the target group compared to the control group. In summary, various studies have shown that miRNAs, including miR-30b, miR-155, miR-181a, and miR-190b, are involved in the proliferation, differentiation, and development of CD4+ T cells. One reason for the lack of increase in CD4+ T cells may be the reduced expression of these miRNAs.