CD4 Values Research Articles

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.

CMV and HIV Coinfection in Women from a Region in Eastern Europe.

(1) Background: Human cytomegalovirus (CMV) infection is one of the most frequent opportunistic infections in immunosuppressed patients. Romania has one of the highest incidences of patients living with human immunodeficiency virus (HIV) which determines an immunosuppressive state. The aim of this study was to establish the prevalence of CMV infection among women living with HIV in Southeastern Romania and also to evaluate and correlate antiretroviral therapy (ART) with CD4 level and CMV disease evolution. (2) Methods: Seventy women living with HIV from Southeastern Romania were screened for CMV infection using antigen quantification. Of these, 50 were included in the study. First, the patients filled out a questionnaire regarding social conditions and other associated diseases. Then, we explored the statistical correlations between the data and HIV status, CD4+ cell counts, viral load, and antiretroviral therapy (ART). (3) Results: Median age of the patients was 33 years. Twenty-nine cases were diagnosed with HIV after sexual life beginning and 21 before. Most of the patients had a CD4 level over 200 cells/µL. ART duration in the CD4 under 200 cells/µL group was a bit longer than that in the CD4 over 200 cells/µL group. Forty-one patients had undetectable viremia. CD4 average value in the lot of patients with undetectable viremia was 704.71 cells/µL and in the lot with detectable viremia was 452.44 cells/µL. Viremia values correlated negatively with CD4 level. A positive correlation between IgG CMV values and ART therapy length was identified. A negative significant correlation between values of IgG CMV and values of CD4 was identified. CD4 value correlated negatively with IgG CMV values and with CMV avidity. (4) Conclusions: IgG CMV values had a weak positive correlation with ART therapy length, and a negative statistically significant correlation with values of CD4. CMV avidity has a negative correlation with CD4 value.

Study of immunophenotypic characteristics, clinicopathological parameters and prognosis in gastric cancer microenvironment

Objective: To explore the immunophenotypic characteristics of gastric cancer microenvironment and analyze its correlation with clinicopathological parameters and prognosis of patients. Methods: The expression levels of leukocyte differentiation antigen (CD) 8, CD4, T lymphocyte immunoglobulin mucoprotein 3 (TIM3), human forkhead box protein P3 (Foxp3) and co-localized tumor infiltrating lymphocytes (TILs) were detected in 92 cases of gastric cancer tissue [58 males and 34 females; aged M(Q1, Q3), 70(59, 77) years ] and 84 cases of paracancer tissue [57 males and 27 females, aged 70(59, 77) years] purchased from Shanghai Xinchao Biotechnology Co., Ltd., and the samples were from 28 hospitals in the sample bank. Gastric cancer and adjacent tissues were divided into high expression group and low expression group according to the optimal cut-off value of positive lymphocytepercentage. The expression of immunophenotypes in gastric cancer and adjacent tissues was analyzed. Kaplan-Meier method was used for survival analysis. Cox proportional hazard model was used to explore the prognostic factors of gastric cancer patients. Results: The optimal cut-off values of CD8, CD4, TIM3 and Foxp3 positive cells in gastric cancer were 12.73%, 1.39%, 10.77% and 2.44%, respectively. The expression of Foxp3 in gastric cancer tissues was higher than that in paracancer tissues [M (Q1, Q3), 0.93 (0.45, 2.16) vs 0.31 (0.09, 0.86), P<0.001], and the expression of CD8 [4.92 (2.34, 8.80) vs 8.81 (6.61, 12.17), P<0.001], CD4 [4.79 (1.77, 11.36) vs 8.40 (4.84, 12.77), P=0.022] and TIM3 [5.68 (2.05, 11.58) vs 7.07 (3.13, 11.43), P=0.338] were lower than that in paracancer tissues. There were significant differences in TIM3 expression in gastric cancer patients with different lymph node metastasis and clinical stage (all P<0.05). The 5-year survival rate of patients with high CD4 expression, low TIM3 expression and low Foxp3 expression in gastric cancer tissues was poor, among which the high CD4 expression and low CD4 expression groups were 29.3% and 64.7%, respectively; The high and low TIM3 expression groups were 60.9% and 30.4%, respectively; The high and low Foxp3 expression groups were 64.3% and 33.3%, respectively (all P<0.05). The optimal cut-off values of CD8+TIM3+TILs, CD4+TIM3+TILs, CD8+Foxp3+TILs and CD4+Foxp3+TILs were 3.86%, 0.23%, 0.08% and 0.76%, respectively. Colocalization analysis showed that the expression of CD8+Foxp3+TILs in gastric cancer tissues was higher than that in adjacent tissues(all P<0.05). Multivariate Cox regression analysis showed that high expression of CD4 (HR=3.079, 95%CI: 1.350-7.024,P=0.008), low expression of TIM3 (HR=0.428, 95%CI: 0.208-0.879, P=0.021) and low expression of Foxp3 (HR=0.288, 95%CI: 0.121-0.687, P=0.005) were the influencing factor for the 5-year survival rate of patients with gastric cancer after operation. Conclusions: Gastric cancer tissues have complex immune microenvironment characteristics. The expression of CD4, TIM3 and Foxp3 is closely related to the prognosis of patients.

Predictive value of CD4+CD8+ double positive T cells for the severity of hemorrhagic fever with renal syndrome

PurposesWe aimed to investigate the levels of CD4+CD8+ double positive (DP) T cells in patients with various severities of hemorrhagic fever with renal syndrome (HFRS), and the predictive capacity of DP T cells for the severity of this disorder. MethodsThe levels of DP T cells in 213 patients and 48 healthy donors were measured by flow cytometry, as were the levels of CD4+ T cells, CD8+ T cells, B lymphocytes, and natural killer (NK) cells. In each type of HFRS patient, we tested the basic clinical reference values for leukocytes, platelets, creatinine (Cr), uric acid (UA), and urea, and the values for activated partial thromboplastin time, prothrombin time, and fibrinogen, using conventional methods. The colloidal gold method was used to measure HFRS antibody levels in the patients. ResultsThe frequency of DP T cells increased with disease severity and peaked in patients with critical disease. Furthermore, the level of DP T cells proportionally correlated with the levels of Cr, UA, and urea in the serum. In contrast, there was an inverse correlation between DP T cells and platelets. Interestingly, the pattern of change in DP T cell frequency was similar to those of CD8+ T cells, B cells, and NK cells, but an inverse tendency was observed for CD4+ T cells. DP T cells demonstrated significant predictive value for the severity of HFRS. ConclusionsThe level of DP T cells is associated with HFRS severity, suggesting that it may be a potent indicator for the course of this disorder.

A major role for CD4+ T cells in driving cytokine release syndrome during CAR T cell therapy

Anti-CD19 chimeric antigen receptor (CAR) Tcell therapy represents a breakthrough for the treatment of B cell malignancies. Yet, it can lead to severe adverse events, including cytokine release syndrome (CRS), which may require urgent clinical management. Whether interpatient variability in CAR Tcell subsets contributes to CRS is unclear. Here, we show that CD4+ CAR Tcells are the main drivers of CRS. Using an immunocompetent model of anti-CD19 CAR Tcell therapy, we report that CD4+, but not CD8+, CAR Tcells elicit physiological CRS-like manifestations associated with the release of inflammatory cytokines. In CAR Tcell-treated patients, CRS occurrence and severity are significantly associated with high absolute values of CD4+ CAR Tcells in the blood. CRS in mice occurs independently of CAR Tcell-derived interferon γ (IFN-γ) but requires elevated tumor burden. Thus, adjusting the CD4:CD8 CAR Tcell ratio to patient tumor load may help mitigate CAR Tcell-associated toxicities.

Prognostic value of CD4&lt;sup&gt;+&lt;/sup&gt; in patients with Т1–2N0M0 breast cancer

Aim. To evaluate the prognostic value of CD4+ in patients with T1–2N0M0 breast cancer.Materials and methods. We performed a quantitative assessment of CD4+ T-lymphocytes in tumor stromal cells and determined the prognostic parameter using the method developed by us (patent No. RU2697709C1 dated 19.08.2019) in 394 patients diagnosed with infiltrative breast cancer. Survival of patients in the analyzed group was assessed over a period of 10 years.Archival material from paraffin blocks of the «target group» was used in the work for molecular genetic profiling techniques (T1–2N0M0, 1118 patients, 2000–2009). A histological preparation stained with hematoxylin and eosin was scanned, and the central and peripheral zones of the tumor were isolated. To identify CD4+ T-lymphocytes, the immunohistochemical study of preparations obtained with rabbit anti-CD4 monoclonal antibodies was performed. After rescanning histological and immunohistochemical preparations, combining different immunohistochemical stains, the cell density in the tumor and stromal components was calculated in preparations stained with hematoxylin and eosin and CD4. For further calculations, we used the data obtained as a result of the number of stromal cells reduced to 1 mm2 (according to the preparation stained with hematoxylin and eosin), the so-called cell density per calculated number of CD4+-stained cells (cell area × proportion of CD4+ cells in percentages). The resulting ratio is used to predict the outcome of breast cancer: if its value is ≤ 50 %, the prognosis is assessed as favorable (in which the patient’s survival is 10 years or more).Results. In 296 (75 %) of 394 observed patients, the proportion of CD4+ T-lymphocytes in tumor stromal cells was ≤ 50 %. Over 10 years, the overall survival of patients in this group was 93 %, which is considered a statistically favorable group for breast cancer (stages T1–2, N0). In 98 (25 %) of 394 observed patients, the proportion of CD4+ T-lymphocytes in tumor stromal cells was ˃50 %. Over 10 years, the overall survival of patients in this group was 82 %, which is considered a statistically unfavorable group for breast cancer (stages T1–2, N0).Conclusion. Our findings suggest high accuracy of our method of breast cancer prognosis based on the quantitative assessment of CD4+ T-lymphocytes in tumor stromal cells. It can be used for breast cancer prognosis in early stages (T1–2, N0).

Psychiatric aspects of healthcare workers during COVID-19 pandemic in Surabaya, Indonesia: prevalence of anxiety, resilience, burnout, and their correlations to stress markers

Link of Video Abstract: https://youtu.be/nI_6kXZ0cSw Introduction: The coronavirus disease (COVID-19) pandemic severely affected healthcare services, including causing negative impacts on healthcare workers’ mental health. An observational study was conducted at Dr. Soetomo General Hospital aimed to measure the prevalence of anxiety, resilience, burnout and their correlations to stress biomarkers amongst healthcare workers in Surabaya, Indonesia, during the COVID-19 pandemic. Methods: Participants were residents who managed COVID-19 patients, chosen using random sampling. All participants had a 1-week shift working with COVID-19 patients in an isolation ward. The study used online questionnaires and laboratory data at the shift’s beginning and end. Along with collecting sociodemographic characteristics, State-Trait Anxiety Inventory (STAI), Brief Resiliency Scale (BRS), and Maslach Burnout Inventory (MBI) scales were administered online. At the same time, laboratory markers such as Neutrophil to lymphocyte ratio (NLR), Interleukin-6 (IL-6), cortisol, and CD4 values were studied. Results: 61 participants were included in the study. At the beginning of the shift, 59% of participants had moderate to severe anxiety, and 83.62% had normal to high resilience. The early proportion of the participants having burnout from combined MBI measures was 88.52%. MBI-EE median scores were lower in the married group (6.50 vs. 9.50, p-value=0.030). NLR value was significantly related to STAI-S and BRS (p-value=0.007 and 0.049, respectively). Conclusions: Most healthcare workers had anxiety and burnout, with normal levels of resilience. Our study found that NLR might be useful as a marker for anxiety and resilience.

Complete blood and absolute CD4+ve T lymphocyte counts of whole blood donors at a hospital blood center in Nigeria

Background: Comprehensive routine screening and adequate selection of the prospective whole blood donor protects his health and safety as well as that of the recipient. Objectives: The main objective of this study was to determine the complete blood and absolute CD4+ T lymphocyte cell counts of apparently healthy prospective whole blood donors. Participants and Methods: This was a hospital based prospective study. A form was designed by the researchers for data collection. The socio demographic status, complete blood cells and absolute CD4+vel counts of apparently healthy whole blood donors who had tested seronegative for HIV, hepatitis B and C markers was captured . Obtained data was analysed with the statistical package for the social scientist software version 20 Results: One hundred male (97.1%) and three female (2.9%) apparently healthy prospective whole blood donors were studied. The median age of study subjects was 30 years. Obtained median blood cell values were 13g/dl, 40%, 4.9/nl and 203.9/nl for haemoglobin concentration, haematocrit, total white cell and platelet counts respectively. The median values for the mean corpuscular haemoglobin concentration (MCHC), mean corpuscular haemoglobin (MCH) and mean corpuscular volume (MCV) of studied blood donors were 32.6g/dl,27.7pg and 85.7fl respectively. Observed prevalence of subnormal blood cell counts for haemoglobin concentration, total white cells and platelets were 12.6%, 25.2%, 13.6% respectively. Also subnormal values for MCHC, MCH, and MCV were 11.7%, 26.2%, 16.5% respectively among studied whole blood donors. No higher than normal blood cell count values were observed. Median values for erythrocyte sedimentation rate and CD4+ T lymphocytes were 8.4mm/hr and 876.2cells/µl respectively. The percentage of subnormal CD4+T lymphocyte count was18.4% Conclusion: A significant percentage of apparently healthy prospective whole blood donors had subnormal blood cell and CD+ve T lymphocyte values. Obtained normal values were comparable with local reference range reports from previous studies in Nigeria and other parts of Africa.

The predictive value of CD4, CD8, and C—reactive protein in the prognosis of schistosomal and non-schistosomal colorectal cancer

BackgroundAlthough schistosomiasis has been basically eliminated, it has not been completely extinction in China and occasional outbreaks occur in Europe in recent years. The relationship between inflammation caused by Schistosoma japonicum and colorectal cancer (CRC) is still obscure, and the inflammation based prognostic systems of schistosomal colorectal (SCRC) has rarely been reported.AimTo explore the different roles of tumor infiltrating lymphocytes (TILs) and C-reactive protein (CRP) in SCRC and in Non-schistosomal CRC (NSCRC), providing a possible predictive system to evaluate outcomes and to improve the risk stratification for CRC patients, especially for CRC patients with schistosomiasis.MethodsThree hundred fifty-one CRC tumors were evaluated for density of CD4 + , CD8 + T cells and CRP in intratumoral and stromal compartments by immunohistochemical using tissue microarray.ResultsThere were no association between TILs and CRP and schistosomiasis. Multivariate analysis identified stromal CD4 (sCD4) (p = 0.038), intratumoral CD8 (iCD8) (p = 0.003), schistosomiasis (p = 0.045) as independent prognostic factors for overall survival (OS) in the whole cohort; and sCD4 (p = 0.006) and iCD8 (p = 0.020) were independent prognostic factors for OS in the NSCRC and SCRC set, respectively. Besides, we found that there were no differences of TILs and CRP, which were distributed in different areas of tumor tissue, between CRC patients with and without schistosomiasis.ConclusionThe results remind us that different subtypes of TILs have distinguished biological behavior and prognosis value in the immune microenvironment of NSCRC and SCRC patients. Meanwhile, the findings require us to stratify patients with schistosomiasis and this might facilitate patient counseling and management.

The Relationship Between Decreased CD-8 T-Cells and Mortality in Patients with COVID-19 Pneumonia in the Intensive Care Unit, A Retrospective Study.

CD-8 T-cells are responsible for the clearance of virally infected cells. In patients with Coronavirus disease-2019 (COVID-19) pneumonia, there are quantitative reductions and functional impairments in T-cells. Low CD-8 T-cell levels cause worse clinical situations. In this study, the relationship between decreased CD-8 T-cells and mortality in patients with COVID-19 pneumonia in the intensive care unit (ICU) was investigated. In this multicenter retrospective study, 277 patients were analyzed. Demographic data, ICU admission scores, blood gas levels, laboratory samples, and outcomes were recorded. Statistical Package for the Social Sciences version 28 was used for statistical analysis. Two hundred forty of 277 patients were included in the study. The mortality rate was 43.3%. In non-survivors, median values of age, Charlson comorbidity index, Acute Physiology and Chronic Health Evaluation II (APACHE-II), procalcitonin, leukocyte count, neutrophil count, neutrophil-lymphocyte count ratio, and duration of invasive mechanical ventilation were significantly higher, whereas median values of PaO2-FiO2 ratio, lymphocyte count, CD-4, and CD-8 T-cells were significantly lower than those in survivors. In the multivariate Cox regression model, the risk of mortality increased 1.04-fold (1.02-1.06) and 1.05-fold (1.01-10.8) by every one unit increase in age and APACHE-II, respectively, whereas it decreased 0.71-fold (0.58-0.87) by every hundred increase in CD-8 T-cells P < 0.001, P=0.007 and P=0.001 respectively. According to our findings, age, APACHE-II, and CD-8 T-cell levels seem to be independent risk factors for mortality in patients with COVID-19 pneumonia in the ICU.

Evaluation of T and B cell immunophenotyping and antibody response post Covid-19 vaccination

PurposeTo evaluate T and B cell subsets and IgG antibodies in response to SARS-CoV-2 post COVID-19 vaccination. MethodsA total of 50 healthy adults (18–60 years) receiving anti-SARS-CoV-2 vaccination (COVISHIELD) were recruited for the study. Blood samples were collected from participants at 3 time points; just before vaccination (Visit 0, V0), just before booster dose (Visit 1, V1) and 6th month after 1st dose (Visit 2, V2). Peripheral blood mononuclear cell isolation was done and evaluated for T and B cell subsets by Flow cytometry. Quantitative determination of IgG antibodies to SARS-CoV-2 was done by Chemiluminescence immunoassay in all samples. Final data for all three visits was available for 37 participants who remained healthy.Ethics approval was obtained from Medanta Institution of Ethics Committee vide MICR No. 1290/2021 dated 24th May 2021. ResultsMean age of the participants was 34.6 ​± ​5.7 years (Range: 24–45 years). Highly significant improvement in SARS-CoV-2 IgG levels was observed after each visit {Mean IgG: (V0 v/s. V1: 133.8 ​± ​339.2AU/ml v/s. 434.5 ​± ​519.2AU/ml; p-value ​= ​0.003) and V0 v/s. V2: 133.8 ​± ​339.2AU/ml v/s. 420.9 ​± ​394.2AU/ml; p-value ​= ​0.002) Between visits 0 and 1, the mean value for CD4 Naïve T cells showed significant increase, while CD4 central memory (CM) T cells showed significant decrease. Between visits 0 and 2 the mean values for CD4 Naïve T cells, CD8 Naïve T cells and Pre germinal centre (Pre GC) B cells showed significant increase. During the same period the mean values for CD4CM, CD8 effector memory (EM) and CD8 CM T cells showed significant decrease. ConclusionIt is concluded that both, humoral and cellular immunity, play an important role in maintaining immunity against COVID-19 infection, following COVISHIELD vaccination. Moreover, in subjects with normalisation of antibody levels post vaccination, persistence of T cell subsets may still offer some immunity.

Characterization of data-driven geriatric syndrome clusters in older people with HIV: a Mexican multicenter cross-sectional study

As living with HIV has been proposed as a condition that may accelerate aging, the main objective of this work was to estimate the prevalence of geriatric syndromes (GS) among older Mexicans with HIV dwelling in the community. Secondly, to evaluate whether the accumulation of GS could be associated with an adverse HIV-related clinical profile, independent of chronological age. Multicenter, cross-sectional study including 501 community-dwelling people aged ≥50 years with HIV. The overall prevalence of nine selected GS and their cumulative number were estimated. An Age-Independent Cumulative Geriatric Syndromes scale (AICGSs) was constructed, and correlations between the AICGSs and HIV-related parameters assessed. Finally, k-mean clustering analyses were performed to test the secondary objective. Median age 56 (IQR: 53-61) years, 81.6% of men. Polypharmacy (74.8%), sensorial deficit (71.2%), cognitive impairment (53.6%), physical disability (41.9%), pre-frailty (27.9%), and falls (29.7%), were the more prevalent GS. A significant negative correlation was found between the AICGSs and normalized values of CD4+ nadir cell counts (r=-0.126; 95%: CI:-0.223 to-0.026, p<0.05). Similarly, a significant inverse adjusted association between the CD4+ nadir cells and the AICGSs was observed on linear regression analysis (β-0.058; 95%: CI:-0.109 to-0.007, p=0.03). Cluster analysis identified three differentiated groups varying by age, metabolic comorbidities, AICGSs, and HIV-related parameters. An elevated prevalence of GS was observed in the studied population. Moreover, the accumulation of GS was associated with adverse HIV-related profiles, independent of age. Thus, early detection and management of GS are crucial to promote healthier aging trajectories in people with HIV. This work was funded in part by the National Center for the Prevention and Control of HIV/AIDS in Mexico (CENSIDA)-National Ministry of Health.

Comparison of peripheral blood T, B, and NK lymphocytes between frontline medical workers for treating patients of COVID-19 and normal outpatient and emergency medical workers in China.

The outbreak of the novel coronavirus disease 2019 (COVID-19) has led to significant mental stress for frontline medical workers treating patients with confirmed COVID-19 in China. Psychological stress has an impact on the immune system. The number and percentage of lymphocyte subsets are standard indicators of cellular immune detection. Here, we reported the differences in CD3, CD4, CD8, CD19, and CD56 lymphocytes between 158 frontline medical workers and 24 controls from medical staffs of the outpatient and emergency departments. We found that frontline medical workers had significantly lower absolute values and percentages of CD19+ B cells, especially in the female and the aged ≥40 years subgroup. Stratification analysis showed that the absolute values of CD4+ T cells were significantly lower in the aged <40 years subgroup, while percentages of CD8+ T cells were lower and percentages of CD56+ NK cells were higher in the aged ≥40 years subgroup. In summary, this study suggests paying more attention to frontline medical workers' mental health and immune function, and properly providing them with psychological interventions and measures of care.

Point of Care CD4 Testing in National Household Surveys - Results and Quality Indicators from Eleven Population-Based HIV Impact Assessment (PHIA) Surveys.

Population-based HIV Impact Assessments (PHIAs) are national household (HH) surveys that provide HIV diagnosis and CD4 testing with an immediate return of results. Accurate CD4 results improve HIV-positive participants' clinical care and inform the effectiveness of HIV programs. Here, we present CD4 results from the PHIA surveys that were conducted in 11 countries in sub-Saharan Africa between 2015 and 2018. All of the HIV-positive participants and 2 to 5% of the HIV-negative participants were offered Pima CD4 (Abbott, IL, USA) point-of-care (POC) tests. The quality of the CD4 test was ensured by conducting instrument verification, comprehensive training, quality control, a review of testing errors and an analysis of unweighted CD4 data by HIV status, age, gender, and antiretroviral (ARV) treatment status. Overall, CD4 testing was completed for 23,085 (99.5%) of the 23,209 HIV-positive and 7,329 (2.7%) of the 270,741 negative participants in 11 surveys. The instrument error rate was 11.3% (range, 4.4% to 15.7%). The median CD4 values among HIV-positive and HIV-negative participants (aged 15+) were 468 cells/mm3 (interquartile range [IQR], 307 to 654) and 811 cells/mm3 (IQR, 647 to 1,013), respectively. Among the HIV-positive participants (aged 15+), those with detectable ARVs had higher CD4 values (508 cells/mm3) than those with undetectable ARVs (385.5 cells/mm3). Among the HIV-positive participants (aged 15+), 11.4% (2,528/22,253) had a CD4 value of less than 200 cells/mm3, and approximately half of them (1,225/2,528 = 48.5%) had detectable ARVs, whereas 51.5% (1,303/2,528) had no detectable ARVs (P < 0.0001). We successfully implemented high quality POC CD4 testing using Pima instruments. Our data come from nationally representative surveys in 11 countries and provide unique insights regarding the CD4 distribution among HIV-positive individuals as well as the baseline CD4 values among HIV-negative individuals. IMPORTANCE The manuscript describes CD4 levels among HIV-positive individuals and baseline CD4 levels among HIV-negative individuals from 11 sub-Saharan countries, thereby highlighting the importance of CD4 markers in the context of the HIV epidemic. Despite increased ARV access in each country, advanced HIV disease (CD4 < 200 cells/mm3) persists among approximately 11% of HIV-positive individuals. Therefore, it is important that our findings are shared with the scientific community to assist with similar implementations of point-of-care testing and to conduct a review of HIV programmatic gaps.

Mass cytometry for the multiplexed quantification and characterization of target expression on circulating cells in whole blood.

Characterization of target abundance on cells has broad translational applications. Among the approaches for assessing membrane target expression is quantification of the number of target-specific antibody (Ab) bound per cell (ABC). ABC determination on relevant cell subsets in complex and limited biological samples necessitates multidimensional immunophenotyping, for which the high-order multiparameter capabilities of mass cytometry provide considerable advantages. In the present study, we describe the implementation of CyTOF® for the concomitant quantification of membrane markers on diverse types of immune cells in human whole blood. Specifically, our protocol relies on establishing Bmax of Ab saturable binding on cells, then converted into ABC according to a metal's transmission efficiency and number of metal atoms per Ab. Using this method, we calculated ABC values for CD4 and CD8 within the expected range for circulating T cells and in concordance with the ABC obtained in the same samples by flow cytometry. Furthermore, we successfully conducted multiplex measurements of the ABC for CD28, CD16, CD32a, and CD64, on >15 immune cell subsets in human whole blood samples. We developed a high-dimensional data analysis workflow enabling semi-automated Bmax calculation in all examined cell subsets to facilitate ABC reporting across populations. In addition, we investigated impacts of the type of metal isotope and acquisition batch effect on the ABC evaluation with CyTOF®. In summary, our findings demonstrate mass cytometry is a valuable tool for concurrent quantitative analysis of multiple targets in specific and rare cell types, thus increasing the numbers of biomeasures obtained from a single sample.

HIV infection and vaccinations

Introduction: Vaccination is mentioned as a most successful way of intervention in public health. Vaccination against other diseases is usually recommended for people with Human Immunodeficiency Virus (HIV/AIDS) infection. Purpose: To identify the correlation of vaccines and vaccination in patients suffering from HIV/AIDS infection. Methodology: This narrative review was based on the bibliographic search of reviews and research studies drawn from the international databases. The exclusion criterion of the articles was the language other than English and Greek. Results: HIV infection is characterized by severe immunodeficiency, which is the result of a decrease in the number of CD4+ T – lymphocytes. The right time to vaccinate a HIV patient is not precisely determined, but it is recommended for those with low CD4 values. All HIV patients are recommended to be vaccinated the annual vaccination of Influenza virus vaccination. It also is recommended to be vaccinated with the vaccination against human papillomavirus (HPV), the vaccination against hepatitis A virus (HAV) and hepatitis B virus (HBV), the vaccination against herpes zoster or shingles and the vaccination against COVID-19 infection. Conclusion: People with HIV/AIDS are more vulnerable to infections from various diseases. Therefore, it is necessary patients with HIV infection be vaccinated against life-threatening diseases.

Therapeutic outcomes of mycophenolate mofetil and glucocorticoid in thyroid-associated ophthalmopathy patients.

To analyze the efficacy of mycophenolate mofetil (MMF) and glucocorticoid administration in patients with thyroid-associated ophthalmopathy (TAO). Sixty patients with moderate to severe TAO treated in Jingzhou Central Hospital from January 2022 to June 2022 were selected and enrtolled in this study. The subjects were divided into experimental group (n=30) and control group (n=30) based on the random number table method. Glucocorticoid pulse therapy was provided in the control group, while MMF was given in the experimental group on the basis of Control group. Clinical activity score (CAS), quality of life (QOL), visual acuity, eyelid fissure width, intraocular pressure, and degree of exophthalmos were observed at the time of admission and at the 12th week and 24th post-treatment weeks. We compared the immune function (TRAb, IL-6, and CD4+/CD8+) of the two groups pre-treatment and 24 weeks post-treatment, and evaluated the clinical therapeutic effect. The clinical effective rates at 12 and 24 weeks in the experimental group were higher (73.3% and 83.3%) than those in the control group (46.7% and 60.0%) (P <0.05). After 12 weeks of treatment, patients' CAS scores, and bilateral lid fissure width decreased and right eye visual acuity increased in the control group compared with those before treatment (P < 0.05); further, after 24 weeks of treatment, patients' QOL scores and bilateral visual acuity increased and CAS scores, bilateral lid fissure width and proptosis decreased compared with those before treatment, and patients' QOL scores, CAS scores and bilateral proptosis improved more than those at 12 weeks of treatment (P <0.05). Additionally, greater improvements were observed in the patients' QOL and CAS scores, and proptosis after 24-week treatment than after 12-week treatment (P<0.05). In the experimental group, the QOL score and binocular visual acuity increased, whereas the CAS score, intraocular pressure, lid width, and proptosis decreased after 12 weeks of treatment as compared to the values of these parameters in the pre-treatment period (P < 0.05); after 24 weeks of treatment, greater improvements were established in the ocular-related indexes improved compared to the pre-treatment period and after 12 weeks of treatment (P < 0.05). After 12 weeks of treatment, the patients in the experimental group had more considerable improvements in the right visual acuity, right intraocular pressure, and left lid fissure width than the control group (P < 0.05); at 24 weeks of treatment, patients in the experimental group had greater improvements in the QOL score, bilateral visual acuity, intraocular pressure, bilateral lid fissure width, and bilateral proptosis than the control group (P < 0.05). No significant differences were found in the values of TRAb, IL-6, and CD4+/CD8+ between the two groups before treatment (P>0.05); the values of TRAb, IL-6, and CD4+/CD8+ in the experimental group was significantly lower than those before treatment and in the control group after 24weeks of treatment. (P>0.05). No statistically significant difference was observed in the incidence of liver damage and menstrual disorders between the two groups during the 24 weeks of treatment (P>0.05). The combination of oral MMF and glucocorticoid shock therapy is an effective drug for the treatment of patients with moderately active TAO.

Analysis of the Efficacy of LNSRH Surgery for Early Cervical Cancer Based on T, B, and NK Lymphocytes

Background: The analysis is based on the level of T, B, NK lymphocytes to evaluate the efficacy of laparoscopic nerve-sparing radical hysterectomy (LNSRH) for early cervical cancer. Methods: Selected 30 patients with early cervical cancer who underwent LNSRH surgery in Tianjin Fifth Central Hospital from December 2019 to January 2021 as the cervical cancer group, and 30 healthy female who underwent physical examination in our hospital during the same period were selected as the control group. The expressions of CD4+, CD8+, CD20+, and CD57+ were detected by flow cytometer, female sexual function index (FSFI) and overall assessment scale of cancer treatment function (FACT-G) score were used to evaluate female sexual function and quality of life, and the receiver operating characteristic curve (ROC) was used to analyze the expression predictive value of CD4+, CD8+, CD20+, and CD57+ in LNSRH of early cervical cancer. Results: The expression of CD4+ and CD57+ in the cervical cancer group was lower than that in the healthy group,and the expression of CD8+ and CD20+ was higher than that in the healthy group (p &lt; 0.05). The expression of preoperative CD4+ and CD57+ was higher for postoperative, and the expression of preoperative CD8+ and CD20+ was lower (p &lt; 0.05) for postoperative. The expression of postoperative 1 day CD4+ and CD57+ was higher for at 3 days postoperative, and the expression of postoperative 1 day CD8+ and CD20+ was lower at 3 days postoperative (p &lt; 0.05). CD4+, CD57+ expression was positively correlated with FSFI score, and CD8+, and CD20+ expression was negatively correlated with FSFI score, CD4+, and CD57+ expression was positively correlated with FACT-G score, and CD8+, and CD20+ expression was negatively correlated with FACT-G score (p &lt; 0.05). ROC analysis of CD4+, CD8+, CD20+, CD57+ expression sensitivity was lower than the four-item combination, the specificity was higher than the four-item combination and the results with significant statistical difference (p &lt; 0.05) and the four combined tests have good predictive value for the efficacy of LNSRH for early cervical cancer (p &lt; 0.05). Conclusions: Cervical cancer patients were mainly cellular immunity and performed low expression of CD4+, high expression of CD8+, high expression of CD20+, and low expression of CD57+. T, B, and NK lymphocytes could be used for immunological detection of cervical cancer.

Comorbidities and New Onset of Type 2 Diabetes Mellitus among Women Living with HIV Receiving Antiretroviral Therapy in Lagos, Nigeria

Background: There is limited gender stratified data in many type 2 diabetes mellitus (T2DM) studies in Africa. This study aimed to determine the prevalence of Comorbidities, New Onset of T2DM and risk factors influencing predisposition to T2DM among women living with HIV (WLWH) and receiving Antiretroviral Therapies. Methods: This was a cross-sectional study of females who were 15 years and above with HIV and/or without pulmonary TB attending HIV and TB DOTS clinics in Lagos, Nigeria from January 2019 to October 2021. The socio-demographic data of participants were obtained using questionnaire. Participants who were not known diabetic cases were tested for diabetes based on WHO and ADA standards using glycated haemoglobin (HbA1c) test, fasting plasma glucose and 2 hour 75 g oral glucose post prandial test. CD4 and CD8 counts were carried out using flow cytometer and their cytokines’ levels were determine using ELISA technique. HIV positive patients with signs and symptoms of TB (presumptive TB) were tested using sputum smear microscopy method and gene X-pert technique. We measured Body mass index (BMI) using International System of Units (kg/m2). Information on age, gender, Antiretroviral therapy (ART), values of CD4, weight, height, viral load and cholesterol test results less than 6 months were obtained from the patients’ folders. Results: A total of 187 WLWH were studied. Their mean age was 41.93±10.32 years. Fifty-five (27.9%) had BMI values of ≥ 30 kg/m2. Twenty-one (11.2%) had impaired glucose level. Those with T2DM at baseline were 5(2.7%), 17(9.1%) later developed T2DM. There was strong association between newly developed T2DM and obesity odd ratio (OR) 8.21(95% CI 1.30-51.99), interleukin (IL-): IL-6 OR 4.50 (95% CI 0.58-35.15), viral load with OR as 1.34(0.48-3.75), knowledge of diabetes OR 2.24 (95% CI 0.43-11.62), consumption of alcohol OR 2.03(95% CI 0.60-6.90). Twenty (10.7%) WLWH developed pulmonary TB while receiving ART. Co-morbidities recorded were: HIV/T2DM 14(7.5%), HIV/TB 21(11.2%), HIV/TB/T2DM 7(3.7%). A total of 42(22.7%) were on 2nd line ART. Conclusion: Prevalence of HIV/TB co-infection, HIV/T2DM comorbidity and HIV/TB/T2DM multimorbidity were high. Also the prevalence of incident TB and new cases of T2DM among the study population were high. There was positive association between development of T2DM while receiving ART and obesity, IL-6, knowledge of diabetes and alcohol consumption. Protease inhibitors were found to influence the development of T2DM among WLWH while receiving ART. Recommendation: There is need for promoting awareness of T2DM and its risk factors among WLWH who are receiving ART.

Inflammatory status in pediatric sickle cell disease: Unravelling the role of immune cell subsets.

Introduction: The mutation of the beta-globin gene that causes sickle cell disease (SCD) results in pleiotropic effects, such as hemolysis and vaso-occlusive crisis that can induce inflammatory mechanisms with deleterious consequences on the organism. Moreover, SCD patients display an increased susceptibility to infections. Few studies are currently available that evaluate a wide immunological profile in a pediatric population. This study proposes an evaluation of the immune profile in subjects with SCD in a pediatric population through a detailed analysis by flow cytometry. Methods and Materials: Peripheral blood samples from 53 pediatric patients with SCD (mean age 9.8years, interquartile range 9years) were obtained and then analyzed by flow cytometry, in order to evaluate changes in the immune populations compared to 40 healthy donors (mean age 7.3years, interquartile range 9.5years). Results: Our data showed an increase in neutrophils (with a reduction in the CD62L + subpopulation) and monocytes (with a decrease in HLA-DRlow monocytes) with normal values of lymphocytes in SCD patients. In the lymphocyte subpopulations analysis we observed lower values of CD4+ T cells (with higher number of memory and central memory T lymphocytes) with increased frequency of CD8+ T cells (with a predominant naive pattern). Moreover, we observed higher values of CD39+ Tregs and lower HLA-DR+ and CD39- T cells with an increased Th17, Th1-17 and Th2 response. Conclusion: We observed immunological alterations typical of an inflammatory status (increase in activated neutrophils and monocytes) associated with a peculiar Treg pattern (probably linked to a body attempt to minimize inflammation intrinsic to SCD). Furthermore, we highlighted a T helper pathway associated with inflammation in line with other studies. Our data showed that immunological markers may have an important role in the understanding the pathophysiology of SCD and in optimizing targeted therapeutic strategies for each patient.