Percentage Of CD34 Research Articles

Effects of blood components of preoperative autologous blood donation on functions of myelopoietic stem cells

ABSTRACTObjectiveTo explore the effects of preoperative autologous whole blood donation and autologous pure red blood cell (RBC) donation on hematopoietic stem cells (HSC), clarify the effects of transfusion by different blood components on HSC, and improve the treatment effect of autotransfusion.MethodsForty healthy male New Zealand rabbits were divided into five groups (n = 8) at random: control (Group A), surgery alone (Group B), surgery + blood sampling (Group C), surgery + preoperative autologous whole blood autotransfusion (Group D), and surgery + preoperative autologous pure RBC autotransfusion (Group E). The rabbits’ bone marrow was collected before surgery (T1), 6 h after surgery (T2), and 24 h after surgery (T3) to detect the CD34+ cell count, the reactive oxygen species (ROS) concentration, the ratio of senescent cells, and the expression of HSC-related proteins (p53 and p21).ResultsCompared with groups A and B, the percentage of CD34+ cells in groups D and E at each time point was significantly increased, while the proportion of senescent cells, ROS, p53 and p21 were significantly decreased (P<0.05). Compared with Group C, the percentage of CD34+ cells at T2 and T3 rose significantly, while the ratio of senescent cells, the ROS concentration, and the content of p53 and p21 declined significantly in Groups D and E (P < 0.05). Compared with Group D, the ratio of senescent cells at T2 declined significantly, the percentage of CD34+ cells at T3 rose significantly, and the ROS concentration at T2 and T3 declined significantly in Group E (P < 0.05).ConclusionFrom the perspective of HSC, collection and transfusion of pure RBC is more beneficial to postoperative recovery of patients than whole blood transfusion.

Characteristics of peripheral lymphocyte subsets in patients with different stages of schistosomiasis japonica.

Immune cells are important for the development of schistosomiasis japonica and are also critical for the treatment of schistosomiasis. The immune cells in the peripheral blood help assess the immune state. The peripheral lymphocytes in schistosomiasis mansoni were well studied; however, immune cells in patients with different stages of schistosomiasis japonica are not well analysed. Here, we performed a preliminary study to explore characteristics of peripheral lymphocyte subsets in patients with different stages of schistosomiasis japonica. 135 patients with Schistosoma japonicum infection and 25 healthy volunteers were included in this study, including 84 patients with chronic S. japonicum infection and 51 patients with advanced S. japonicum infection. Flow cytometry analysis was performed to evaluate peripheral lymphocytes including T cells, B cells, and natural killer (NK) cells. Blood routine and liver function test data were analysed. Ultrasound examination was used to access liver fibrosis according to the World Health Organization standard about ultrasound in schistosomiasis. Demographic data analysis suggested there was no difference in age and gender in patients with S. japonicum infection and health control group. Liver function tests showed that patients with advanced schistosomiasis had a higher incidence of liver function abnormality and blood lipid than those with chronic schistosomiasis. Blood routine results reflected that haemoglobin, red blood cells, platelets, as well as lymphocytes in the advanced group were significantly less than that in the chronic group. Furthermore, flow cytometry analysis indicated that the percentage of CD4+ T cells was lower in the advanced group, but the percentage of CD19+ B cells was higher in the advanced group. In addition, the number of CD3+ T cells, CD3+ CD4+ T cells, CD3+ CD8+ T cells, and NK cells was less in the advanced group when compared with those in the chronic group. In addition, there was a correlation between the decrease in CD4+ T cells and more severe fibrosis on ultrasound images. Our results indicated that the immune state in the peripheral is different in different stages of S. japonicum infection. Lymphocyte subset analysis has potential to facilitate differential diagnosis of different stages of schistosomiasis japonica and even to be a prognostic factor.

Elevated circulating CD19+CD24hiCD38hi B cells display pro-inflammatory phenotype in idiopathic membranous nephropathy

CD19+CD24hiCD38hi regulatory B cells exert immunosuppressive functions by producing IL-10, but their role in idiopathic membranous nephropathy (IMN) remains elusive. Here, we investigated the frequency and functional changes of circulating CD19+CD24hiCD38hi B cells and evaluated the correlation of CD19+CD24hiCD38hi B cells with clinical features and T helper cell subsets in IMN patients. Compared with healthy controls (HCs), IMN patients showed an increased frequency of CD19+CD24hiCD38hi B cells, but a significant reduction in the percentage of CD19+CD24hiCD38hi B cells was observed 4 weeks after cyclophosphamide treatment. The frequency of CD19+CD24hiCD38hi B cells was positively correlated with the levels of 24h urinary protein, but negatively correlated with serum total protein and serum albumin, respectively. CD19+CD24hiCD38hi B cells in IMN patients displayed a skewed pro-inflammatory cytokine profile with a higher level of IL-6 and IL-12, but a lower concentration of IL-10 than their healthy counterparts. Accompanied by upregulation of Th2 and Th17 cells in IMN patients, the percentage of CD19+CD24hiCD38hi B cell subset was positively associated with Th17 cell frequency. In conclusion, CD19+CD24hiCD38hi B cells were expanded but functionally impaired in IMN patients. Their altered pro-inflammatory cytokine profile may contribute to the pathogenesis of IMN.

Investigating the relationship between the number and activity of natural killer cells with increased cytomegalovirus and CMV disease after kidney transplantation

BackgroundCytomegalovirus (CMV) infections caused by the cytomegalovirus are one of the most common problems in patients after kidney transplant. We examined the association of the relationship between the number and activity of natural killer cells with increased cytomegalovirus and its related disease after kidney transplantation. Material and methodsIn this analytical study, 58 new transplant patients in the Labbafinejad Hospital, who did not have any evidence of CMV infection, were evaluated based on the number and percentage of CD56+/16+, CD56+/16−, and CD69+ Natural Killer (NK) cells. ResultsThe results of this study showed that CD16+ and CD56+ cells in the group of CMV Ag-positive patients are less than negative patients (p = 0.003) and the difference between the two groups are significant (p = 0.01). However, CD69+ cells did not differ significantly between the two groups (p = 0.1). Moreover, the absolute number of CD16+ and CD56+ cells declined significantly after infection with CMV unlike the CMV Ag - group(p = 0.003). DiscussionThese results indicate that kidney transplant patients suffering from CMV infection after transplantation have a significantly reduced total number of NK cells. On the other hand, a slight decrease in the number of NK subgroups was observed with an increase in the peak serum levels of cyclosporine. As a consequence of these findings, it can be assumed that more dosage and a higher level of the drug will result in more severe immunosuppression and, consequently, increased susceptibility to CMV infections. Thus, taking the right dose of the drug would prevent viral infections and immune system from over-activation.

Macrophages Provide Essential Support for Erythropoiesis, and Extracellular ATP Contributes to a Erythropoiesis-Supportive Microenvironment during Repeated Psychological Stress.

Psychological stress is a significant contributor to various chronic diseases and affects multiple physiological processes including erythropoiesis. This study aimed to examine the tissue-specific contributions of macrophages and extracellular ATP, as a signal of disturbed tissue homeostasis, to erythropoiesis under conditions of repeated psychological stress. Adult male BALB/c mice were subjected to 2 h daily restraint stress for seven consecutive days. Clodronate-liposomes were used to deplete resident macrophages from the bone marrow and spleen two days prior to the first restraint procedure, as well as newly recruited macrophages, every third day for the duration of the experiment. Repeated stress induced a considerable increase in the number of erythroid progenitor cells as well as in the percentage of CD71+/Ter119+ and CD71-/Ter119+ cells in the bone marrow and spleen. Macrophage depletion completely abolished the stimulative effect of repeated stress on immature erythroid cells, and prevented stress-induced increases in ATP levels, P2X7 receptor (P2X7R) expression, and ectonucleotidase CD39 activity and expression in the bone marrow and spleen. The obtained results demonstrate the stimulative effects of repeated stress on erythroid cells, extracellular ATP levels, P2X7R expression, CD39 activity and expression within the bone marrow and spleen, as well as the essential role of macrophages in stress-induced changes.

Liver Cancer Vascularity Driven by Extracellular Matrix Stiffness: Implications for Imaging Research.

Extracellular matrix stiffness represents a barrier to effective local and systemic drug delivery. Increasing stiffness disrupts newly formed vessel architecture and integrity, leading to tumor-like vasculature. The resulting vascular phenotypes are manifested through different cross-sectional imaging features. Contrast-enhanced studies can help elucidate the interplay between liver tumor stiffness and different vascular phenotypes. This study aims to correlate extracellular matrix stiffness, dynamic contrast-enhanced computed tomography, and dynamic contrast-enhancement ultrasound imaging features of 2 rat hepatocellular carcinoma tumor models. Buffalo-McA-RH7777 and Sprague Dawley (SD)-N1S1 tumor models were used to evaluate tumor stiffness by 2-dimensional shear wave elastography, along with tumor perfusion by dynamic contrast-enhanced ultrasonography and contrast-enhanced computed tomography. Atomic force microscopy was used to calculate tumor stiffness at a submicron scale. Computer-aided image analyses were performed to evaluate tumor necrosis, as well as the percentage, distribution, and thickness of CD34+ blood vessels. Distinct tissue signatures between models were observed according to the distribution of the stiffness values by 2-dimensional shear wave elastography and atomic force microscopy ( P < 0.05). Higher stiffness values were attributed to SD-N1S1 tumors, also associated with a scant microvascular network ( P ≤ 0.001). Opposite results were observed in the Buffalo-McA-RH7777 model, exhibiting lower stiffness values and richer tumor vasculature with predominantly peripheral distribution ( P = 0.03). Consistent with these findings, tumor enhancement was significantly greater in the Buffalo-McA-RH7777 tumor model than in the SD-N1S1 on both dynamic contrast-enhanced ultrasonography and contrast-enhanced computed tomography ( P < 0.005). A statistically significant positive correlation was observed between tumor perfusion on dynamic contrast-enhanced ultrasonography and contrast-enhanced computed tomography in terms of the total area under the curve and % microvessel tumor coverage ( P < 0.05). The stiffness signatures translated into different tumor vascular phenotypes. Two-dimensional shear wave elastography and dynamic contrast-enhanced ultrasonography adequately depicted different stromal patterns, which resulted in unique imaging perfusion parameters with significantly greater contrast enhancement observed in softer tumors.

Tumor immune environment in colitis-associated colorectal cancers: Comparison with sporadic colorectal cancers.

e15531 Background: Persistent inflammatory diseases such as ulcerative colitis (UC) and Crohn’s disease (CD) contribute to the tumor initiation and progression in colitis-associated cancers (CAC). Intestinal damage caused by persistent inflammation promotes clonal expansion of somatic and epithelial cells and alters the immune microenvironment. A comprehensive investigation in comparing the tumor immune environment in CAC with sporadic colorectal cancers (CRC) will provide more clinical usefulness of immunoprofiling in CRC. Methods: Thirteen patients with CAC (6 with UC and 7 with CD) and 10 patients with sporadic colorectal cancer from our hospital were enrolled in the study. FFPE samples from patients were analyzed for the tumor infiltrating lymphocytes [PD-L1+, CD3+, CD4+, CD8+, CD20+, CD68+, CD163+] by multiplex immunofluorescence technique. The core and peritumor areas annotated by pathologists on images. Then artificial intelligence (AI) algorithms delineated the tumor-enriched segments and stroma, and calculated the density and percentage of each marker in two regions. A nonparametric approach (Kruskal–Wallis) was used to compare the density and percentage of immune cells between CAC and sporadic colorectal cancer groups. Results: In the stroma area, the density (counts per mm2) of PD-L1+CD68+ cells in patients with CAC were significantly higher than those in sporadic cancer patients (p &lt; 0.05). However, the density of PD-L1+CD68+ did not show significant differences related to disease types (CAC or sporadic CRC) in the tumor-enriched segments. Significantly, the density of CD3+CD4+ cells and the percentage of CD68+CD163- cells in patients with CAC showed a trend of higher density than those in sporadic cancer patients in the stroma area (all, 0.05 &lt; p &lt; 0.1). The density and the percentage of CD20+ cells in CAC patients had a higher density tendency than those in sporadic cancer patients both in the tumor-enriched segments and stroma (all, 0.05 &lt; p &lt; 0.1). Other indicators in tumor-enriched segments and stroma, such as CD3+, CD4+, CD8+, CD68+CD163+ and PD-L1+, had no significant difference between CAC and sporadic CRC patients, regardless of the percentage or the density. Conclusions: We identified that the tumor immune environment of CAC was different from that of sporadic CRC. Some tumor infiltrating lymphocytes such as PD-L1+CD68+ cells infiltrated more in CAC than those in sporadic cancer, suggesting that CAC may respond better to immunotherapy than sporadic CRC. However, this hypothesis requires a further validation with a larger sample size.

Shiga Toxin, Stx2e, Influences the Activity of Porcine Lymphocytes In Vitro.

Oedema disease (OD) in piglets is one of the most important pathologies, as it causes significant losses due to the high mortality because of the Shiga toxin family, which produces Escherichia coli (STEC) strains. The main toxin responsible for the characteristic pathologies in pigs is Shiga toxin 2 subtype e (Stx2e). Moreover, there is growing evidence that Stx's family of toxins also targets immune cells. Therefore, this study evaluated the effect of different concentrations of Stx2e on porcine immune cells. Porcine peripheral blood mononuclear cells were pre-incubated with Stx2e, at three different concentrations (final concentrations of 10, 500, and 5000 CD50/mL) and with a negative control group. Cells were then stimulated with polyclonal mitogens: concanavalin A, phytohemagglutinin, pokeweed mitogen, or lipopolysaccharides. Cell proliferation was assessed by BrdU (or EdU) incorporation into newly created DNA. The activation of the lymphocyte subsets was assessed by the detection of CD25, using flow cytometry. The toxin significantly decreased mitogen-driven proliferation activity, and the effect was partially dose-dependent, with a significant impact on both T and B populations. The percentage of CD25+ cells was slightly lower in the presence of Stx2e in all the defined T cell subpopulations (CD4+, CD8+, and γδTCR+)-in a dose-dependent manner. B cells seemed to be the most affected populations. The negative effects of different concentrations of Stx2e on the immune cells in this study may explain the negative impact of the subclinical course of OD.

Gingipains are the important virulence factors of Porphyromonas gingivalis downregulating B10 cells.

Porphyromonas gingivalis (P. gingivalis) is a keystone pathogen in periodontitis. Our previous study indicated that periodontitis induced by P. gingivalis increased the percentage of CD19+ B cells, but decreased the ratio of IL-10 producing regulatory B cells (B10) in collagen-induced arthritis (CIA) mice. It's still unclear which virulence factors of P. gingivalis are involved in these processes. Here, we compared the effects of different components of P. gingivalis on the biogenesis of B10 cells, and found that the decreased proportion of B10 cells was mainly resulted from the undenatured proteins other than the DNA, RNA or LPS of P. gingivalis. As gingipains are enzymes and virulence factors which play vital role in the progression in periodontitis through affecting innate and adaptive immune system, we then compared the influence of the wild-type (WT) strain of P. gingivalis (ATCC 33277) and its isogenic gingipain-null mutant (∆K∆RAB) on differentiation of splenic B cells into B10 cells. Interestingly, compared to WT strain, ∆K∆RAB treatment increased the frequency of B10 cells as well as expression of IL-6 in B cells. Furthermore, the acute peritonitis, an ideal model for quick evaluation of immune effects of agents, induced by ∆K∆RAB showed higher IL-6 production and proportion of B10 cells compared with WT. Finally, we performed transcriptomic analysis to better understand the effects and possible mechanism of gingipains on B cells. Compared with WT, ∆K∆RAB upregulated the PI3K-Akt pathway of B cells which is important for IL-10 production and B10 cell biogenesis, and more activated Jak-STAT pathway which is a classical signaling pathway mediated by IL-6. Cumulatively, this study preliminarily revealed that gingipains of P. gingivalis are vital virulence factors downregulating B10 cells and altering immune responses. This article is protected by copyright. All rights reserved.

Characterization of lymphocyte profiles in children with syndromic obesity

BackgroundLittle is known about blood lymphocyte subpopulations in children with common (CO) or syndromic (SO) obesity. We aimed to describe the blood lymphocyte profiles of obese children and to search for associations with clinical phenotypes. MethodsMain blood lymphocyte subpopulations were analyzed in 159 children with CO and 34 with SO in a retrospective cohort. Phenotypes included obesity history, body mass index (BMI) Z score, percentage fat mass, and inflammatory parameters. Correlations were performed between phenotypes and circulating lymphocyte profiles. ResultsChildren with SO had a higher BMI Z score (5.5 ± 1.7 SD) than children with CO (4.7 ± 0.9 SD; p = 0.01). Significant differences were found for lymphocyte counts, including a higher percentage of CD19+ B cells (SO = 20.1 ± 6.7 vs. CO = 17.1 ± 6.1%, p = 0.03), despite lower absolute numbers (SO = 0.57 ± 0.20 vs. CO = 0.63 ± 1.9 g/L, p < 0.01). However, no difference in the lymphocyte profile was found between children with SO and those with the most severe CO (BMI Z score ≥ 4.7 SD). ConclusionChildren with SO have altered blood lymphocyte profiles with increased prevalence of CD19+ B cells, which is closely linked to the degree of obesity severity and inflammatory markers.

Abstract 4449: The expression and immune regulation of thymic stromal lymphopoietin in non-small cell lung cancer development and progression

Abstract Purpose: Previous studies have shown thymic stromal lymphopoietin (TSLP) to be an important driver of type 2 inflammation. A new and unexpected function for TSLP has been found in the induction and progression of a variety of tumors. This study conducted TSLP performance in lung cancer patients and the correlation of the clinical pathological data. Materials and Methods: The patients with pathological diagnosis of lung cancer were enrolled. The percentage of CD19 + B cells was determined in the lymphocyte gate and counted via flow cytometry. Relevant serum cytokines including TSLP were analyzed using LEGENDplex software. In vitro study was conducted by A549 cell line and the mRNA expression change of TSLP was extracted by Reverse Transcriptase PCR (RT-PCR). Results: When comparing the frequencies of B cell subsets between early-stage (N = 31) and late-stage lung cancer group (N = 29), the frequency of memory B cell (17.3±6.1% vs 23.4±15.1%, p=0.042), CD27+IgM+ B cell (1.8±1.3% vs 2.9±2.3%, p=0.030), class-switched B cell (15.5±5.5% vs 22.2±14.9%, p=0.023), class-switched memory B cell (69.9±11.3% vs 59.9±19.7%, p=0.017) and plasmablast (8.5±7.3% vs 18.0±20.8%, p=0.019) were significantly different between groups. The patients with higher frequency of class-switched B cell had significantly worse prognosis than low frequency (HR 3.054, 95% CI 1.007-9.262, p=0.049). The level of IL-1B, IL-6, IL-10, IL-12, IL-13, and TSLP were significantly higher in the late-stage cancer patients. Simulation in vitro test with human lung A549 cell line was performed, the expression of short-form TSLP (sfTSLP) level decreased with the higher anti-cancer drug as Pemetrexed concentration. Conclusions: The increase proportion of CD27+CD38- (class-switched) B cell could be an independent, poor prognostic factor for NSCLC patients. TSLP signaling is addressed to the anti-cancer therapeutic response. Therefore, TSLP and relevant cytokines may be involved directly and indirectly in shaping the inflammatory status of the tumor microenvironment. Citation Format: Chung-Yu Chen, Ying-Yin Chen, Hung-Chueh Peng, Yi-Ling Ye. The expression and immune regulation of thymic stromal lymphopoietin in non-small cell lung cancer development and progression. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4449.

Determination of the relationship between CD34+ stem cell amount and DMSO in hematopoetic stem cell transplantation

It is important to calculate the CD34+ stem cell (SC) count at the right time in patients with hematological malignancies who will undergo Hematopoietic Stem Cell Transplantation (HSCT). The amount of SC infused into the patient affects the engraftment time and healing process of the patient. In this study, we aimed to compare which of the DMSO-not removed and DMSO-removed samples showed the CD34 + SC amount more accurately as the SC amount determination method after the SC was dissolved after cryopreservation in patients who will undergo HSCT. A total of 22 patients were included in the study. All 22 patients were transplanted from frozen samples using DMSO. After the SC products were dissolved in a 37 °C water bath, they were washed 2 times and the amount of CD34+ SC was studied from the samples taken by removing DMSO and without removing DMSO. In the findings, the amounts of CD34+ SC studied with both methods were compared. The increase in the number and percentage of CD34+ SC after DMSO-removed was found to be statistically significant both in terms of difference and proportionally, and the calculated effect sizes also showed that the increase was clinically significant (Cohen's d is between 0.43 and 0.677). After thawing the frozen SCs of the patients who will undergo HSCT, the analysis of CD34+ SCs from which DMSO is removed provides a more accurate calculation of the CD34+ SC amount in the AP.

DNA Damage Response (DDR) Is Associated With Treatment-free Remission in Chronic Myeloid Leukemia Patients.

DNA Damage Response (DDR) Is Associated With Treatment-free Remission in Chronic Myeloid Leukemia Patients.

COVID-19 vaccine uptake, hesitancy and clinical effects on patients with Takayasu's arteritis: A web-based questionnaire survey from a large cohort.

This study aimed to investigate the Coronavirus disease 2019 (COVID-19) vaccination rate, reasons for vaccine hesitancy and clinical effects on patients with Takayasu's arteritis (TAK). A web-based survey was administered to a TAK cohort established by the Department of Rheumatology, Zhongshan Hospital through WeChat in April, 2022. Responses from a total of 302 patients were received. The Sinovac or Sinopharm inactivated vaccination rate, side effects, and vaccine hesitancy reasons were analyzed. In addition, disease flare, new disease onset, and changes of immune-related parameters after vaccination were analyzed in vaccinated patients. Among 302 patients, 93 (30.79%) received the inactivated COVID-19 vaccination. Among the 209 unvaccinated patients, the most common reason for hesitancy were concern about side effects (136, 65.07%). Vaccinated patients had a longer disease duration (p = 0.08) and lower use of biologic agents (p < 0.001); 16 (17.20%) of the 93 vaccinated patients developed side effects, and most of them were mild; 8 (8.60%) developed disease flares or new-onset disease 12-128 days post-vaccination and 2 (2.15%) developed serious adverse effects (vision defect and cranial infarction). Immune-related parameters of 17 patients indicated decreases in IgA and IgM after vaccination (p < 0.05). Eighteen (19.35%) of the 93 vaccinated patients were diagnosed post-vaccination.These patients had a significantly higher percentage of CD19+ B cells at disease onset (p < 0.05) than the unvaccinated patients diagnosed at the same time. The vaccination rate was low in TAK, which was mainly caused by concerns about negative effects of vaccination on their disease. An acceptable safety profile was observed in vaccinated patients. The risk of disease flare associated with COVID-19 vaccination warrants further investigation.

The Role of Cluster of Differentiation 39 (CD39) and Purinergic Signaling Pathway in Viral Infections.

CD39 is a marker of immune cells such as lymphocytes and monocytes. The CD39/CD73 pathway hydrolyzes ATP into adenosine, which has a potent immunosuppressive effect. CD39 regulates the function of a variety of immunologic cells through the purinergic signaling pathways. CD39+ T cells have been implicated in viral infections, including Human Immunodeficiency Virus (HIV), Cytomegalovirus (CMV), viral hepatitis, and Corona Virus Disease 2019 (COVID-19) infections. The expression of CD39 is an indicator of lymphocyte exhaustion, which develops during chronicity. During RNA viral infections, the CD39 marker can profile the populations of CD4+ T lymphocytes into two populations, T-effector lymphocytes, and T-regulatory lymphocytes, where CD39 is predominantly expressed on the T-regulatory cells. The level of CD39 in T lymphocytes can predict the disease progression, antiviral immune responses, and the response to antiviral drugs. Besides, the percentage of CD39 and CD73 in B lymphocytes and monocytes can affect the status of viral infections. In this review, we investigate the impact of CD39 and CD39-expressing cells on viral infections and how the frequency and percentage of CD39+ immunologic cells determine disease prognosis.

The Change in Subpopulation and Function of Natural Killer Cells in Adult Patients with Infectious Mononucleosis Related Liver Injury

To investigate the changes of subpopulation and function of natural killer (NK) cells in adult patients with infectious mononucleosis related liver injury. Seventeen healthy controls, twenty-six patients of infectious mononucleosis (IM) without liver injury, and twenty-one IM patients with liver injury were enrolled in this study. Peripheral blood was collected. Plasma and peripheral blood mononuclear cells (PBMC) were purified. Plasma interleukin-15 (IL-15) level was measured by enzyme linked immunosorbent assay. Three NK cell subpopulations, including CD56+CD16-, CD56+CD16+, and CD56-CD16+ NK cells, were assessed by flow cytometry. PBMC were co-cultured with 721.221, K562, and P815-Ab cells, respectively. CD107a expression and CD16 mean fluorescence intensity (MFI) in NK cells was investigated by flow cytometry. PBMC from IM patients with liver injury were stimulated with anti-IL-15 neutralizing antibody, and the change of NK cell function was observed. One-way ANOVA and paired t test were used for statistical analysis. The percentage of CD56+CD16- NK cells in IM patients with liver injury (7.36±0.92)% and IM patients without liver injury (10.67±1.37)% were lower than that of controls (12.02±2.51)% (P<0.05). CD56+CD16- NK cell frequency in IM patients with liver injury also was lower than that in IM patients with liver injury (P<0.0001). The percentage of CD56+CD16+ NK cells in IM patients with liver injury (62.03±8.78)% and IM patients without liver injury (44.74±13.03)% were higher than that in controls (40.74±10.15)% (P<0.05). CD56+CD16+ NK cell frequency in IM patients with liver injury also was higher than that in IM patients with liver injury (P<0.0001). There was no statistical difference of CD56-CD16+ NK cell proportion among three groups (P=0.427). Plasma IL-15 level in IM patients with liver injury (309.1±68.00) pg/ml were higher than that in IM patients without liver injury (269.6±53.57) pg/ml and controls (257.4±73.93) pg/ml (P<0.05). NK cells induced target cell death through different receptors upon activation, the ratio of CD107a-positive cells in NK cells of IM patients with liver injury was higher than that in IM patients without liver injury and controls (P<0.05), while CD16 MFI in NK cells in IM patients with liver injury was lower than that in IM patients without liver injury and controls (P<0.05). The percentage of CD107a positive cells in NK cells was significantly reduced in response to anti-IL-15 neutralizing antibody stimulation in IM patients with liver injury (P<0.05). IM patients with liver injury presented imbalance of NK cell subpopulation and enhanced NK cell function. The elevation of IL-15 might contribute to promote NK cell cytotoxicity in IM patients with liver injury.

The density of bone marrow mononuclear cells and CD34+ cells in patients with three neurologic conditions

BackgroundThis study aimed to identify the density of mononuclear cells (MNCs) and CD34+ cells in the bone marrow of patients with three neurologic conditions.MethodsThe study included 88 patients with three neurologic conditions: 40 with cerebral palsy (CP) due to oxygen deprivation (OD), 23 with CP related to neonatal icterus (NI), and 25 with neurological sequelae after traumatic brain injury. Bone marrow aspiration was conducted from the patients’ bilateral anterior iliac crest under general anesthesia in an operating theater. MNCs were isolated by Ficoll gradient centrifugation and then infused intrathecally.ResultsThere was a significant difference in the average MNC per ml and percentage of CD34+ cells by the type of disease, age group, and infusion time (p value < 0.05). The multivariable regression model showed the percentage of CD34+ association with the outcome (gross motor function 88 items- GMFM-88) in patients with CP.ConclusionsThe density of MNCs was 5.22 million cells per mL and 5.03% CD34+ cells in patients with three neurologic conditions. The highest density of MNCs in each ml of bone marrow was found in patients with CP due to OD, whereas the percentage of CD34+ cells was the highest among patients with CP related to NI.

The quality and quantity media-cultured mononuclear cell transplantation is safe and effective in ischemic hindlimb mouse model

ObjectiveThis study was conducted to investigate in vitro proangiogenic and anti-inflammatory phenotypes and functions and the in vivo efficacy and safety of quality and quantity (QQ) media-cultured mononuclear cells (MNCs) compared with standard cultured MNCs from the peripheral blood of patients with chronic limb-threatening ischemia (CLTI) with atherosclerotic risk factors. MethodsPeripheral blood MNCs (PBMNCs) from patients with CLTI were cultured in QQ culture media or standard culture media. Phenotypic analysis of progenitor cells (CD34+CD133+), M2 macrophages (CD206+), and inactivated T regulatory cells (CD4+CD25+CD127+), colony-forming assay, and tube formation assay of QQ media-cultured MNCs (QQMNCs) and PBMNCs, were conducted. Intramuscular transplantation of QQMNCs or PBMNCs was performed in the ischemic hindlimb model. The clinical appearance of ischemic limbs was observed, and blood flow in ischemic limbs was measured using a laser Doppler perfusion imager. Outcomes were compared between the QQMNC and PBMNC groups. ResultsTwenty patients with CLTI were included. The mean percentages of CD34+ cells, CD133+ cells, CD34+CD133+ progenitor cells, CD206+ cells, colony-forming cells, and tube formation were significantly higher in the QQMNCs. The mean percentage of CD4+CD25+CD127+ cells was significantly lower in QQMNC. The colony-forming unit count and Dil-acetylated low-density lipoprotein uptake were significantly greater in QQMNCs. The clinical appearance of post-QQMNC-injected limbs was less severe than the appearance of post-PBMNC-injected limbs. Limb perfusion was significantly better in the QQMNCs. ConclusionsProangiogenic and anti-inflammatory phenotypes of MNCs cultured in QQ culture media were reproducible. Intramuscular QQMNC transplantation was safe and resulted in better reperfusion of ischemic hindlimbs compared with PBMNCs.

Efficacy and safety of belimumab for the treatment of refractory childhood-onset systemic lupus erythematosus: A single-center, real-world, retrospective study.

This study aimed to investigate the efficacy and safety of belimumab for treating children with refractory childhood-onset systemic lupus erythematosus (cSLE). Twenty-six cSLE patients who received belimumab treatment in our hospital from January 2020 to September 2021 (23 of them for more than 52 weeks) were enrolled in this study. Their clinical and laboratory data, assessment of disease activity, glucocorticoid dosage, and treatment-emergent adverse events (TEAEs) were retrieved for analysis. The paired samples t-test and the nonparametric test were used to compare the baseline and post-treatment data. The mean age of onset was 10.3 ± 2.4 years old; the mean disease duration was 41.6 ± 37.4 months; the median Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K) score was 10 (P 25, P 75: 3, 17); and the mean Physician's Global Assessment (PGA) score at baseline was 1.9 ± 1.0. Compared with the baseline values, there was a significant decrease in the 24-h urine protein quantifications at 24 and 52 weeks of treatment (P<0.05) as well as an elevated complement (C) 3 and C4 levels at 4, 12, 24, and 52 weeks of treatment. In addition, the SLEDAI-2K and PGA scores as well as the percentage of CD19+ B cells were significantly decreased at 12, 24, and 52 weeks of treatment compared with the baseline values (P<0.05). The dosage of glucocorticoid at 4, 12, 24, and 52 weeks of treatment was significantly less than that at baseline or the previous follow-up (P<0.05). At 52 weeks, 14 subjects (53.8%) achieved Lupus Low Disease Activity State (LLDAS), and 4 subjects (15.4%) reached clinical remission (CR). At the last follow-up, 16 subjects (61.5%) achieved LLDAS, and 10 subjects (38.5%) reached CR. Belimumab treatment can significantly improve laboratory indicators, reduce disease activity, and decrease the dosage of glucocorticoid required in children with cSLE. Moreover, it has a good safety profile.

Reduced immune system responsiveness in fibromyalgia - A pilot study

Fibromyalgia is characterized by widespread musculoskeletal and joint pain, stiffness, fatigue, and sleep and mood disorders. However, the involvement of the immune system in the pathways of fibromyalgia is unclear. The aim of this study was to explore the role of the immune system in comparison with healthy controls and in association with clinical symptoms. Thirteen women with fibromyalgia and 14 controls were included. Peripheral blood mononuclear cells were stimulated and analysed by flow cytometry, and interferon gamma (IFN-γ) and interleukins were measured. Among clinical symptoms, the fibromyalgia group showed decreased cold pain threshold. Immunologically, they had a higher percentage of monocytes, a lower percentage of CD19 + B - cells, and lower secretion of IFN-γ after stimulation. Decreased capacity to secrete IFN-γ was significantly correlated with decreased cold pain threshold in the fibromyalgia group. These results confirm the presence of immune aberrations in fibromyalgia, at least partially responsible for the associated pain.