In-depth human immune cellular profiling from newborn to frail.

Immune functional decline and remodeling accompany aging and frailty. It is still largely unknown how changes in the immune cellular composition differentiate healthy individuals from those become frail at a relatively early age. Our aim in this exploratory study was to investigate immunological changes from newborn to frailty, and the association between health statute and various immune cell subtypes. The participants analyzed in this study covered human cord blood cells and peripheral blood cells collected from young adults, healthy and frail old individuals. A total of 30 immune cell subsets was performed by flow cytometry based on the surface markers of immune cells. Furthermore, frailty was investigated for its relations with various leukocyte subpopulations. Frail individuals exhibited a higher CD4/CD8 ratio, a higher proportion of CD4+ central memory T (TCM) cells, CD8+ effector memory T cells, CD27- switched memory B (CD27-BSM) cells, CD27+ switched memory B cells, age-associated B cells (ABCs) and CD38-CD24- B cells, and a lower proportion of naïve CD8 + T cells and progenitor B cells. The Frailty index score was found to be associated with naïve T cells, CD4/CD8 ratio, ABCs, CD27-BSM cells, and CD4+ TCM cells. Our findings conducted a relatively comprehensive and extensive atlas of age- and frailty-related changes in peripheral leukocyte subpopulations from newborn to frailty. The immune phenotypes identified in this study can contribute to a deeper understanding of immunosenescence in frailty and may provide a rationale for future interventions and diagnosis.

Cellular infiltration, cytokines, and histopathology of skin lesions associated with different clinical forms and stages of naturally occurring lumpy skin disease in cattle.

Lumpy skin disease (LSD) caused by the Capripoxvirus LSD virus which infects cattle, leading to a serious disease characterized by fever and the eruption of skin nodules all over the surface of the body. Our understanding of the pathogenesis of this disease is still incomplete, particularly the immunopathological alterations occurring in the skin nodules of infected animals. Therefore, we collected skin nodules from naturally infected cattle with different forms of the disease, both in the early stage of clinical infection and after disease progression. The skin samples were examined both histopathologically and immunohistochemically using a variety of antibodies targeting immune cellular markers and cytokines. As a result, the dermatohistopathology revealed orthokeratotic hyperkeratosis, vasculitis, epidermal microvesicles, and cellules claveleuses of Borrel in the early stage of infection, with the severity of the lesions correlating with the severity of the clinical disease. Meanwhile, late-stage samples had epidermal hyperkeratosis as well as dermal lymphocytic and histiocytic infiltrations. The predominant cellular infiltrates in the cutaneous lesions of early-stage LSD samples were interferon (IFN)-γ+ cells and CD4+ T lymphocytes with few macrophage lineage cells. However, in the late-stage samples, numerous Iba-1+ macrophages, with few IFN-γ+ cells and CD4+ T lymphocytes, were detected. Our findings indicate that IFN-γ+ cells, CD4+ T lymphocytes, and macrophages play a key role in the immunity against natural LSD virus infection and imply that cutaneous vasculopathy associated with LSD virus infection is an immune-mediated lesion. The current study contributes to our understanding of the pathogenesis of LSD.

Curcumin Hybrid Lipid Polymeric Nanoparticles: Antioxidant Activity, Immune Cellular Response, and Cytotoxicity Evaluation.

Poor solubility and short biological half-life present a challenge that needs to be overcome in order to improve the recognized bioactivities of curcumin (CUR), the main phenolic compounds derived from the roots of Curcuma longa. However, drug delivery systems have proven to be an excellent strategy to improve and obtain greater bioavailability. Our previous studies on curcuminoid hybrid nanoparticles have shown promising results by significantly increasing the solubility of desmethoxycurcumin (DMC) and bisdemethoxycurcumin (BDM). In this contribution, we performed a detailed characterization of a CUR as well as in vitro and in vivo studies. The developed method produced CUR loaded nanoparticles with an average size of 49.46 ± 0.80. Moreover, the FT-IR analysis confirmed the encapsulation, and TEM images showed their spherical shape. The NP achieved an encapsulation efficiency greater than 99%. Further, the release studies found that the NPs obtained a significantly higher release than the pure compounds in water. In vivo delayed-type hypersensitivity (DTH) studies showed promising results by enhancing the immune activity response of CUR in NP compared to bulk CUR. Furthermore, we report a significant increase in antioxidant activity for CUR-NP in aqueous solution compared to free CUR. Finally, an important in vitro cytotoxic effect on gastric AGS and colon SW620 adenocarcinoma cell lines was found for CUR-NP while empty carrier nanoparticles are observed to exhibit low cytotoxicity, indicating the potential of these CUR-PLU NPs for further studies to assess their phytotherapeutic applications.

Tumor Microenvironment Characterization in Breast Cancer and an Immune Cell Infiltration Score Development, Validation, and Application.

The tumor microenvironment (TME) refers to the cellular environment in which tumors exist. An increasing number of reports have emphasized its role in tumor progression, prognosis, relapse, metastasis, and therapeutic response with breast cancer (BRCA). Few studies have revealed a systematic landscape of immune cell infiltration (ICI) in BRCA. In this study, we comprehensively analyzed the immune cells infiltrating TME in BRCA. Three ICI patterns were identified through an unsupervised clustering method and an ICI score was developed by a principal component analysis (PCA). A Kaplan-Meier survival with log-rank test revealed a significant overall survival (OS) difference of BRCA patients with these three ICI patterns. We also found that a high ICI score was characterized by an elevated tumor mutation burden (TMB), effector T-cell infiltration, INF-γ-related cytotoxicity, and cytolytic activity score. An independent cohort validated that this ICI score could be a prognostic indicator for BRCA. Two immunotherapeutic cohorts and two chemotherapeutic cohorts confirmed that patients with higher ICI scores showed significant chemotherapeutic and immunotherapeutic advantages. In summary, these results suggest that the ICI patterns could act as a prognostic indicator and that the ICI score could precisely predict the clinical outcome for BRCA patients.

In-depth immune cellular profiling reveals sex-specific associations with frailty.

BACKGROUND: With advancing age, the composition of leukocyte subpopulations in peripheral blood is known to change, but how this change differs between men and women and how it relates to frailty is poorly understood. Our aim in this exploratory study was to investigate whether frailty is associated with changes in immune cell subpopulations and whether this differs between men and women. Therefore, we performed in-depth immune cellular profiling by enumerating a total of 37 subpopulations of T cells, B cells, NK cells, monocytes, and neutrophils in peripheral blood of 289 elderly people between 60-87 years of age. Associations between frailty and each immune cell subpopulation were tested separately in men and women and were adjusted for age and CMV serostatus. In addition, a random forest algorithm was used to predict a participant's frailty score based on enumeration of immune cell subpopulations. RESULTS: In the association study, frailty was found to be associated with increased numbers of neutrophils in both men and in women. Frailer women, but not men, showed higher numbers of total and CD16- monocytes, and lower numbers of both CD56+ T cells and late differentiated CD4+ TemRA cells. The random forest algorithm confirmed all the findings of the association studies in men and women. In men, the predictive accuracy of the algorithm was too low (5.5%) to warrant additional conclusions on top of the ones derived from the association study. In women however, the predictive accuracy was higher (23.1%), additionally revealing that total T cell numbers and total lymphocyte numbers also contribute in predicting frailty. CONCLUSIONS: In-depth immune cellular profiling revealed consistent associations of frailty with elevated numbers of myeloid cell subpopulations in both men and women. Furthermore, additional associations were found between frailty and lower numbers of some T cell subpopulations, in women only. Thus, our study indicates sex-specific associations of immune subpopulations with frailty. We hope that our study will prompt further investigation into the sex-specific immune mechanisms associated with the development of frailty.

Umbilical cord mesenchymal stem cell modulates immune cell subsets in lupus mice / 中华风湿病学杂志

Objective The purpose of this study is to observe the changes of immune cell subsets in lupus mice after umbilical cord mesenchymal stem cells (UC-MSCs) transplantation. Methods B6.MRL-Faslpr lupus mice were randomly divided into the following three groups: the UC-MSCs treated group, the fibroblast like synoviocytes (FLS) treated group and the untreated group. MSC (1×106) or FLS (1×106) were injected into the tail vein of lupus mice respectively. Four weeks after treatment, the spleen index was calculated. The pathological changes of kidney were assessed by HE staining. The frequencies of immune cell subsets in spleen and macrophage in kidney as well as abdominal cavity were analyzed by flow cytometry. Data were analyzed with t test. Results The spleen index of UC-MSCs treated lupus mice [(79 ±9) mg/10 g] and IgG level [(7.5±1.5) mg/ml] were significantly decreased when compared with FLS treated group [(147±23) mg/10 g, t=2.78, P<0.01] [(17.0 ±2.8) mg/ml, t=3.00, P<0.01] and the untreated group [(156 ±16) mg/10 g, t=4.29, P<0.01] [(16.7 ±1.6) mg/ml,t=4.01, P<0.01]. HE staining also showed that the pathological changes of kidney were alleviated after MSCs transplantation. In addition, the frequency of plasma cells in the untreated group [(2.61 2± 0.318)% vs (0.306±0.017)%, t=7.22, P<0.01] and the FLS treated group [(2.412±0.297)% vs (0.306±0.017)%, t=7.07, P<0.01] were markedly higher than MSCs treatment [(0.306 ±0.017)%]. Moreover, the frequency of CD25+Foxp3+/CD4+Treg in the MSCs treated group [(15.08±0.81)%] was significantly increased compared with the untreated group [(8.02 ±0.47)%, t=7.45, P<0.01] and FLS treated group [(8.80 ±0.23)%, t=7.39, P<0.01]. MSCs treatment resulted in a decrease in CXCR5+PD1+/CD4+Tfh and IFNγ+/CD4+Th1 subset, compared with the untreated group [(14.3±1.5)%vs (31.5±3.3)%, t=5.25, P<0.01] [(1.78±0.27)% vs (5.93±1.56)%, t=2.60, P<0.05] and the FLS treated group [(14.3±1.5)%vs (28.8±2.2)%, t=5.49, P<0.01] [(1.78±0.27)%vs (4.88±0.81)%, t=3.61, P<0.01]. The frequency of macrophage in kidney of the MSCs treated group [(3.52 ±0.37)%] was markedly increased compared with the untreated group[(1.58±0.29)%, t=3.25, P<0.01], while neither the IL4+/CD4+Th2 subset nor the IL17+/CD4+Th17subset and the frequency of macrophage in abdominal cavity showed significant changes in the three groups. Conclusion These findings suggest that the therapeutic effects of MSCs on lupus mice may mediate through increasing the frequency of spleen Treg and renal macrophage and decreasing the frequency of Tfh, Th1 and plasma cells.

Interferon-beta affects mitochondrial activity in CD4+ lymphocytes: Implications for mechanism of action in multiple sclerosis.

BACKGROUND: Whereas cellular immune function depends on energy supply and mitochondrial function, little is known on the impact of immunotherapies on cellular energy metabolism. OBJECTIVE: The objective of this paper is to assess the effects of interferon-beta (IFN-ß) on mitochondrial function of CD4(+) T cells. METHODS: Intracellular adenosine triphosphate (iATP) in phytohemagglutinin (PHA)-stimulated CD4(+) cells of multiple sclerosis (MS) patients treated with IFN-ß and controls were analyzed in a luciferase-based assay. Mitochondrial-transmembrane potential (ΔΨm) in IFN-ß-treated peripheral blood mononuclear cells (PBMCs) was investigated by flow cytometry. Expression of genes involved in mitochondrial oxidative phosphorylation (OXPHOS) in CD4(+) cells of IFN-ß-treated individuals and correlations between genetic variants in the key metabolism regulator PGC-1α and IFN-ß response in MS were analyzed. RESULTS: IFN-ß-treated MS patients exhibited a dose-dependent reduction of iATP levels in CD4(+) T cells compared to controls (p < 0.001). Mitochondrial effects were reflected by depolarization of ΔΨm. Expression data revealed changes in the transcription of OXPHOS-genes. iATP levels in IFN-ß-responders were reduced compared to non-responders (p < 0.05), and the major T allele of the SNP rs7665116 of PGC-1α correlated with iATP-levels. CONCLUSION: Reduced iATP-synthesis ex vivo and differential expression of OXPHOS-genes in CD4(+) T cells point to unknown IFN-ß effects on mitochondrial energy metabolism, adding to potential pleiotropic mechanisms of action.

Cellular immune function changes and effect of thymosin alpha-1 on the changes in elderly patients with severe pneumonia / 中华老年医学杂志

Objective To investigate the cellular immune function changes and the effect of thymosin alpha-1 on the changes in elderly patients with severe pneumonia. Methods T cell subset and natural killer (NK) cell were detected in 66 elderly patients with severe pneumonia and 34 elderly patients with common pneumonia. The severe pneumonia patients were randomly divided into 2 groups: the treatment groups (34 cases) and the control group (32 cases). All patients received conventional therapy of pneumonia. The treatment group received 1.6 mg of thymosin alpha-1 through subcutaneous injection once a day for a week and twice a week later. Results The levels of CD3, CD4, CD8 and NK cell were lower in elderly patients with severe pneumonia than in patients with common pneumonia [(43.54%±18.97%) vs. (45.46%±10.43%), (25.43%±12.72%) vs. (38.47%±8.20%), (16.68%±9.30%) vs. (22.36%±8.06%), (13.52%±4.66%) vs. (17.87%±7.11%), t=-6.779、-5.85、-3.161、-3.285 respectively all P＜0.05]. The levels of CD3, CD4, CD4/CD8 and NK cell increased significantly after treatment in treatment group [(64.22%±5.53%) vs. (61.53%±13.41%), (31.70%±4.38%) vs. (26.07%±4.31%), (1.27%±0.91%) vs. (0.97%±0.22%), (17.67%±4.56%) vs. (15.44%±3.82%), F=5.591,11.526,8.934,4.564 respectively, all P＜0.05]. The duration of antibiotic injection and length of stay were lower in treatment group than in control group [(14.17±2.51) d vs. (14.42±2.79) d, (12.69±2.80) d vs. (15.04±3.58) d, t=-3.152、-2.690 respectively, all P＜0.05]. Conclusions The immune function of the elderly patients with severe pneumonia is lower. Thymosin alpha-1 can improve the immune function of the elder patients with severe pneumonia and is helpful for controlling an infection.

Effect of immune modulation therapy on heart function and cytokines in elder patients with chronic heart failure / 中华老年医学杂志

Objective To investigate the effect of immune modulation therapy on heart function and cytokines in elder patients with chronic heart failure (CHF). Methods The 96 patients aged 60-78 years with New York Heart Association(NYHA)functional. class Ⅱ-Ⅳ CHF were randomly divided into two groups: CHF treatment group received regular therapy and thymopetidum and CHF control group received regular therapy. Another 45 healthy individuals aged 60-80 years were involved as normal control. The ejection faction of left ventricle (LVEF), inflammatory cytokines including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), anti-inflammatory cytokine interleukin-10 (IL-10), plasma high sensitive C-reactive protein (hsCRP), plasma brain natrium peptide (BNP)and Minnesota Living with Heart Failure Questionnaire (LHFQ) assessment were tested before therapy, 15 days and 75 days after treatment. Results (1) Before therapy, compared with normal control group, the levels of TNF-α, IL-1β, TNF-α/IL-10 ratio, BNP, hsCRP and LHFQ were significantly increased (P ＜ 0. 05 or P ＜ 0. 01 ), and the levels of IL-10, LVEF were markedly decreased (P＜0.01) in the patients of CHF treatment group and CHF control group. While no difference between the two CHF groups was observed. (2) After the first course of treatment,compared with CHF control group, the levels of IL-10 were increased (P＜0. 01), while the levels of TNF-α, IL-1β, BNP and hsCRP were decreased (P＜0.05 or P＜0.01) in CHF treatment group. The level of LVEF was increased, TNF-α/IL-10 ratio (4.84 ±0. 53 vs. 5.28±0. 66) and LHFQ were decreased even though there was no significant difference between the two groups. (3) After the second course of treatment, compared with CHF control group, the levels of IL-10 and LVEF were increased (P＜0. 05 or P＜0.01), while the level of TNF-α, IL-1β, TNF-α/IL-10 ratio (4.55±0. 69 vs. 5.18±0.38), BNP, hsCRP and LHFQ were decreased (P＜0.05 or P＜0.01) in CHF treatment group. Conclusions Thymopetidum, as an immunemodulating agent, might regulate the equilibrium of cytokines and improve the heart function of patients with CHF, indicating that immune modulation therapy might improve the treatment strategy for CHF patients.

Cellular immune responses of recombinant hepatitis B(rHB)vaccine and HBsAg derived from Hansenular polymorpha cells / 中华流行病学杂志

Objective To study the kinetics of immune response in mice and human immunized with rHB vaccine or rHBsAg derived from yeast cells(Hansenula polymorpha).Methods With different doses,the level of IFN-γ secreted by spleen mononuclear cells(MNC)including CD8+T cells by MACs of mice were detected by enzyme-linked immunospot(ELISPOT)methods after stimulation in vitro with HBsAg MHC class Ⅰ peptide S28-39,respectively.At serial time points.the immunized mice were detected for IFN-γ by ELISPOT as above and for the lymphocytotoxicity test(CTL)by specific lysis assay.The levels of IFN-γ,IL-2,IL-5 and anti-HBs in mice induced by rHB vaccine were detected after single or three doses.Four adults were vaccinated with rHB vaccine according to 0,1 and 2 month schedule.The peripheral blood mononuclear cells(PBMCs)were collected at the 3,8,21,34 and 65 days after the first dose.The CD8+T cells with high purity obtained by sorting from PBMcs were stimulated with rHBsAg or HBsAg peptides.The SFC of IFN-γ,IL-2 and IL-4 of CD4+ and CD8+ T cells were determined by ELISPOT.Results The cytokine of IFN-γ became detectable on day 7 and its peak value appeared on day 14 by ELISPOT.The CTL was detected on day 7 and the maximum lysis of CTL appeared on day 28.The cellular immune response of IFN-γ of MNCs were significantly correlated with the doses vaccinated from 1 μg to 8 μg(Υpositive rates=0.951,Ppositive rates=0.049＜0.05;rSFC=0.996,PSFC=0.000＜0.05).IFN-γSFC of CD8+T cells were significantly associated with the doses from 1 μg to 4 μg(Υ=0.999,P=0.025＜0.05).The HBsAg specific cellular immune and humoral responses of mice immunized with three doses were significantly higher than that with a single dose(P＜0.05).The characteristics of IFN-γ,IL-2 and IL-4 of CD4+ and CD8+ T cells were variable between individuals immunized with the same rHB vaccine.The level of IL-2 and IL-4 of responders were significantly related to the titer of anti-HBs.Conclusion Data from this study showed the kinesis of cellular immunity in mice and adults vaccinated with rHBsAg or rHB vaccine respectively.and the characteristics of cellular immune response in adults induced by the vaccine.Our data provided the basis of standardizing the analysis of cellular immune response to rHB vaccine.

Comparative effects of autologous blood transfusion on human immune function and recovery in lung cancer patients / 中国基层医药

Objective To investigate the change in natural kill cell,T-ly mphocyte subset counts and effect of recovery after homologous and autologous blood transfusion.Methods 50 patients undergoing lung cancer operation were randomly divided into two groups:group A received autologous blood transfusion with 400ml and group H received homologous blood transfusion with 400ml.Venous blood samples were taken before surgery,before blood transfusion and on the 1st and 5th postoperative day for determination of T-ly mphocyte subsets and natural kill cell counts by flow cytomet.The existent ratio of 1～3 years were followed up.Results The NK cell,CD+_3 and CD+_4 counts and CD+_4/CD+_8 ratio decreased significantly on the 1st postoperative day in both groups(P

Study of immune function of peripheral blood dendritic cells from chronic hepatitis B patients / 中华传染病杂志

Objective To study immune function of peripheral blood dendritic cells in chronic hepa titis B virus infected patients. Methods Peripheral blood DCs of patients and normal human were isolated and cultured in serum free media. The expression levels of DC surface molecule were analyzed by flow cytometry and the ability of DC to induce T lymphocyte proliferation were evaluated by a liquid scintillation counter and the amounts of cytokines in MLR were measured detected. Results The expression rate of CD86 (70.2?5.2)% on DC in patients was decreased compared with that in controls. (95.3?3.5)%, P

Study of perioperative enteral immunonutrition in patients with gastrointestinal cancer / 中国现代普通外科进展

Objective:To evaluate the effects of perioperatively administrated enteral immunonutrition in gastrointestinal cancer patients on immune and inflammatory responses,nutrition states and postoperative morbidity.Methods:Sixty patients with gastrointestinal cancer were divided randomly into two groups.Two groups perioperatively(from preoperative day 5 to postoperative day 7) received an supplemented diet with arginine,RNA,and ?-3fatty acids(immunonutrition group;n=30) or an isoenergetic and isonitrogenous standard diet (standard-nutrition group;n=30).All variables of immune and inflammatory responses,nutrition states and postoperative morbidity were measured on preoperative day 5 and postoperative day 1,4 and 8.Results:On postoperative day 4 and 8,most immune variables and prealbumin in the immuno-group were significantly higher than those in the standard-group(P0.05),and inflammatory variables as CRP in the immunonutrition group were significantly lower than those in the standard group.In the immunonutrition group,there were significantly fewer patients who experienced postoperative complications and shorter days of hospital stay compared with standard group(P0.05).Conclusion:The perioperative administration of enteral immunonutrition in gastrointestinal cancer patients can significantly modulate the postoperative immunosuppressive and inflammatory responses at the early postoperative day,can significantly decrease the occurrence of infectious and overall postoperative complications as well as the length of hospital stay.