Reference values of lymphocyte subsets from healthy Polish adults.

The flow cytometry method could support physicians' decisions in the diagnosis and treatment monitoring of immunodeficient patients. Most clinical recommendations are focused on the search for alterations in T- and B-lymphocyte subsets, less commonly natural killer (NK) cells and granulocytes. While reference values for clinically meaningful lymphocyte subsets have been published ubiquitously among numerous countries, we have not found significant data for a population of adult Polish habitats; thus we determined reference values for T, B, and NK subsets according to sex and age. The female group showed a higher percentage of lymphocytes (CD45++), T helper lymphocytes with a higher absolute count, as well as CD4/CD8 ratio, marginal zone-like B cells, class-switched B cells, and CD21low B cells than the male group. The male group was found to have elevated percentages of naïve B lymphocytes, transitional B cells, and plasmablasts. A weak positive correlation with age was found among double positive T lymphocytes, natural killer T cells (NKT) lymphocytes, and CD21low B cells. A negative correlation with age for double negative T lymphocytes, marginal zone-like B cells, and plasmablasts was noted. The results indicated the importance of creating distinct reference ranges regarding sex and age concerning immunophenotype.

Ruxolitinib inhibits IFNγ-stimulated Sjögren's salivary gland MSC HLA-DR expression and chemokine-dependent T cell migration.

OBJECTIVES: Sjögren's disease (SjD) is a systemic autoimmune disease characterized by focal lymphocytic infiltrate of salivary glands (SGs) and high SG IFNγ, both of which are associated with elevated lymphoma risk. IFNγ is also biologically relevant to mesenchymal stromal cells (MSCs), a SG resident cell with unique niche regenerative and immunoregulatory capacities. In contrast to the role of IFNγ in SjD, IFNγ promotes an anti-inflammatory MSC phenotype in other diseases. The objective of this study was to define the immunobiology of IFNγ-exposed SG-MSCs with and without the JAK1 & 2 inhibitor, ruxolitinib. METHODS: SG-MSCs were isolated from SjD and controls human subjects. SG-MSCs were treated with 10 ng/ml IFNγ +/- 1000 nM ruxolitinib. Experimental methods included flow cytometry, RNA-sequencing, chemokine array, ELISA and transwell chemotaxis experiments. RESULTS: We found that IFNγ promoted expression of SG-MSC immunomodulatory markers, including HLA-DR, and this expression was inhibited by ruxolitinib. We confirmed the differential expression of CXCL9, CXCL10, CXCL11, CCL2 and CCL7, initially identified with RNA sequencing. SG-MSCs promoted CD4+ T cell chemotaxis when pre-stimulated with IFNγ. Ruxolitinib blocks chemotaxis through inhibition of SG-MSC production of CXCL9, CXCL10 and CXCL11. CONCLUSIONS: These findings establish that ruxolitinib inhibits IFNγ-induced expression of SG-MSC immunomodulatory markers and chemokines. Ruxolitinib also reverses IFNγ-induced CD4+ T cell chemotaxis, through inhibition of CXCL9, -10 and -11. Because IFNγ is higher in SjD than control SGs, we have identified SG-MSCs as a plausible pathogenic cell type in SjD. We provide proof of concept supporting further study of ruxolitinib to treat SjD.

The correlation between γδ-T lymphocyte subset in early postnatal peripheral blood and brain injury of preterm infants / 中华实用儿科临床杂志

Objective:To investigate the correlation between preterm infants with brain injury and the proportion of lymphocyte subsets, especially γδ-T cells in the postnatal peripheral blood, and to determine the predictive potential of γδ-T cells in the early peripheral blood in brain injury.Methods:It was a prospective study involving 106 preterm infants with gestational age less than 34 weeks who were delivered in the Department of Neonatology, the Third Affiliated Hospital of Zhengzhou University from January 1, to June 1, 2021.Relative levels of γδ-T , CD4 + T, CD8 + T, CD3 + T and total lymphocyte subsets in peripheral blood collected within the first 24 hours after birth were measured by flow cytometry.Recruited infants were divided into brain injury group (36 cases) and non-brain injury group (70 cases) according to serial cranial ultrasound and magnetic resonance imaging(MRI) at the corrected gestational age of 36-37 weeks.Differences in general conditions and the proportion of lymphocyte subsets between groups were compared by the t-test or Chi- square test.Patients in brain injury group were further divided into intracranial hemorrhage(ICH) group(8 cases), periventricular leukomalacia (PVL) group (6 cases)and diffuse white matter damage (WMD) group(22 cases). The proportion of lymphocyte subsets among the different groups was compared by One- Way ANOVA, followed by the LSD- t test. Results:The proportion of γδ-T cells in postnatal peripheral blood of preterm infants at 24 hours after birth in brain injury group was significantly lower than that of non-brain injury group [(0.09±0.12)% vs.0.15±0.13)%, t=-2.445, P=0.016]. No significant differences in the proportion of the CD4 + and CD8 + T cell subsets were found between them.Both preterm infants in PVL group and WMD group had a significantly lower proportion of γδ-T cells at 24 hours after birth compared to that of the non-brain injury group [(0.03±0.05)%, (0.07±0.09)% and (0.15±0.13)%], respectively, ( t=-2.190, -2.659, all P<0.05). Conclusions:γδ-T cells in early postnatal peripheral blood may be involved in the development of brain injury in preterm infants and they had early predictive value for preterm infants at high risk of brain injury, especially the leukomalacia and diffuse white matter injury.

Relationship between CD8 +FoxP3 +CD25 + T cell subsets and the therapeutic effect of pembrolizumab in patients with uterine cervical cancer / 肿瘤研究与临床

Objective:To investigate the relationship between CD8 +FoxP3 +CD25 + T cell subsets and the therapeutic effect of programmed death receptor 1 (PD-1) inhibitor pembrolizumab in treatment of uterine cervical cancer. Methods:The data of 105 patients with uterine cervical cancer who received pemblizumab therapy based on chemotherapy in the First Hospital of Qinhuangdao from January 2018 to January 2020 were retrospectively analyzed. Flow cytometry was used to detect the ratio of CD8 +FoxP3 +CD25 + T cell in peripheral blood of patients. The efficacy and safety were analyzed. According to the efficacy, all patients were divided into remission group (complete remission + partial remission) and non-remission group (stable disease + progressive disease). The clinical characteristics and CD8 +FoxP3 +CD25 + T cell ratio of the two groups were compared. Multivariate logistic regression model was used to analyze the influencing factors for the efficacy. The efficacy of CD8 +FoxP3 +CD25 + T cell ratio predicting the therapeutic effect of patients was analyzed by using receiver operating characteristic (ROC) curve. Results:The objective remission rate of all patients was 17.14% (18/105), and the incidence of adverse reaction was 39.05% (41/105). The proportion of patients with a family history of cervical cancer in the remission group was lower than that than in the non-remission group [5.56% (1/18) vs. 34.48% (30/87)], and the difference was statistically significant ( χ2=6.00, P=0.014). The proportion of CD8 +FoxP3 +CD25 + T cell of 105 patients before and after treatment was (0.83±0.21)% and (0.77±0.10)%, respectively; the proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the remission group was (0.55±0.26)%, (0.31±0.12)%, respectively; the proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the non-remission group was (0.89±0.30)%, (0.87±0.28)%, respectively. The proportion of CD8 +FoxP3 +CD25 + T cell after treatment in the remission group was lower than that before treatment ( P < 0.05); there was no statistically significant difference in the proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the non-remission group ( P>0.05). The proportion of CD8 +FoxP3 +CD25 + T cell before and after treatment in the non-remission group was higher than that in the remission group (all P<0.001). The proportion of CD8 +FoxP3 +CD25 + T cell higher than the mean value of both groups before treatment and the proportion of CD8 +FoxP3 +CD25 + T cell higher than the mean value of both groups after treatment were independent risk factor of disease remission ( OR=2.542, 95% CI 1.649-3.918, P<0.001; OR=2.936, 95% CI 2.154-4.002, P<0.001). ROC curve analysis showed that the area under the curve of CD8 +FoxP3 +CD25 + T cell ratio predicting the disease remission before treatment was 0.720, and its best cut-off value was 0.77%, the senfitivity was 77.78%, the specificity was 70.11%. Conclusions:Early detection of CD8 +FoxP3 +CD25 + T cell ratio helps to predict the effect of PD-1 inhibitor pembrolizumab therapy for uterine cervical cancer.

Lymphocytes subsets in correlation with clinical profile in CVID patients without monogenic defects.

Objectives: Common variable immunodeficiency (CVID) patients experience clinical manifestations rather than recurrent respiratory infections including autoimmunity, enteropathy, and lymphoproliferation. We evaluated the correlation of lymphocyte subpopulations with such manifestations.Methods: Twenty-six genetically unsolved CVID patients were subdivided into four phenotypes: infection only (IO), autoimmunity (AI), chronic enteropathy (CE), and lymphoproliferative disorders (LP) and examined for lymphocyte subsets by flow cytometry and TCD4+ proliferation by Carboxyfluorescein succinimidyl ester (CFSE) test.Results: We detected reduced memory B and increased total, effector memory (EM), cytotoxic, and activated TCD8+ in IO, AI and CE, decreased plasmablasts, total and naive TCD4+, Regulatory TCD4+ (Treg) and naive TCD8+ in IO and CE, elevated CD21low B and terminally differentiated effector memory (TEMRA) TCD8+ in IO and AI, increased helper T (Th2) and Th17 in IO, decreased Th1 in AI and defective total and naive B and central memory (CM) TCD4+ in CE. IO showed reduced TCD4+ proliferation response.Conclusions: In genetically unsolved CVID patients, increased Th2 and Th17 and reduced Treg is associated with IO, increased CD21low B and TEMRA TCD8+ and reduced Th1 is contributed to AI and reduced total and naive B, CM TCD4+ and naive TCD8+ and expanded total TCD8+ is correlated with CE.

The Cellular Composition of the Innate and Adaptive Immune System Is Changed in Blood in Response to Long-Term Swimming Training.

Competitive swimming requires high training load cycles including consecutive sessions with little recovery in between which may contribute to the onset of fatigue and eventually illness. We aimed to investigate immune changes over a 7-month swimming season. Fifty-four national and international level swimmers (25 females, 29 males), ranging from 13 to 20 years of age, were evaluated at rest at: M1 (beginning of the season), M2 (after the 1st macrocycle's main competition), M3 (highest training load phase of the 2nd macrocycle) and M4 (after the 2nd macrocycle's main competition) and grouped according to sex, competitive age-groups, or pubertal Tanner stages. Hemogram and the lymphocytes subsets were assessed by automatic cell counting and by flow cytometry, respectively. Self-reported Upper Respiratory Symptoms (URS) and training load were quantified. Although the values remained within the normal range reference, at M2, CD8+ decreased (M1 = 703 ± 245 vs. M2 = 665 ± 278 cell µL-1; p = 0.032) and total lymphocytes (TL, M1 = 2831 ± 734 vs. M2 = 2417 ± 714 cell µL-1; p = 0.007), CD3+ (M1 = 1974 ± 581 vs. M2 = 1672 ± 603 cell µL-1; p = 0.003), and CD4+ (M1 = 1102 ± 353 vs. M2 = 929 ± 329 cell µL-1; p = 0.002) decreased in youth. At M3, CD8+ remained below baseline (M3 = 622 ± 245 cell µL-1; p = 0.008), eosinophils (M1 = 0.30 ± 0.04 vs. M3 = 0.25 ± 0.03 109 L-1; p = 0.003) and CD16+56+ (M1 = 403 ± 184 vs. M3 = 339 ± 135 cell µL-1; p = 0.019) decreased, and TL, CD3+, and CD4+ recovered in youth. At M4, CD19+ were elevated (M1 = 403 ± 170 vs. M4 = 473 ± 151 cell µL-1; p = 0.022), CD16+56+ continued to decrease (M4 = 284 ± 131 cell µL-1; p < 0.001), eosinophils remained below baseline (M4 = 0.29 ± 0.05 109 L-1; p = 0.002) and CD8+ recovered; monocytes were also decreased in male seniors (M1 = 0.77 ± 0.22 vs. M4 = 0.57 ± 0.16 109 L-1; p = 0.031). The heaviest training load and higher frequency of URS episodes happened at M3. The swimming season induced a cumulative effect toward a decrease of the number of innate immune cells, while acquired immunity appeared to be more affected at the most intense period, recovering after tapering. Younger athletes were more susceptible at the beginning of the training season than older ones.

The underlying changes and predicting role of peripheral blood inflammatory cells in severe COVID-19 patients: A sentinel?

BACKGROUND: The underlying changes of peripheral blood inflammatory cells (PBICs) in COVID-19 patients are little known. Moreover, the risk factors for the underlying changes of PBICs and their predicting role in severe COVID-19 patients remain uncertain. MATERIAL AND METHODS: This retrospective study including two cohorts: the main cohort enrolling 45 patients of severe type serving as study group, and the secondary cohort enrolling 12 patients of no-severe type serving as control group. The PBICs analysis was based on blood routine and lymphocyte subsets. The inflammatory cell levels were compared among patients according to clinical classifications, disease-associated phases, as well as one-month outcomes. RESULTS: Compared with patients of non-severe type, the patients of severe type suffered from significantly decreased counts of lymphocytes, eosinophils, basophils, but increased counts of neutrophils. These PBICs alterations got improved in recovery phase, but persisted or got worse in aggravated phase. Compared with patients in discharged group, the patients in un-discharged/died group suffered from decreased counts of total T lymphocytes, CD4 + T lymphocytes, CD8 + T lymphocytes, as well as NK cells at 2 weeks after treatment. Clinical classification-critically severe was the independently risk factor for lymphopenia (OR = 7.701, 95%CI:1.265-46.893, P = 0.027), eosinopenia (OR = 5.595, 95%CI:1.008-31.054, P = 0.049), and worse one-month outcome (OR = 8.984; 95%CI:1.021-79.061, P = 0.048). CONCLUSION: Lymphopenia and eosinopenia may serve as predictors of disease severity and disease progression in COVID-19 patients, and enhancing the cellular immunity may contribute to COVID-19 treatment. Thus, PBICs might become a sentinel of COVID-19, and it deserves attention during COVID-19 treatment.

[Clinical characteristics and T-lymphocyte subsets in 48 acquired immune deficiency syndrome patients with cytomegalovirus infections].

Objective: To investigate the clinical features and T lymphocytes subsets in patients with acquired immune deficiency syndrome (AIDS) and cytomegalovirus (CMV) infection. Methods: A total of 48 hospitalized patients with human immunodeficiency virus (HIV)-1/AIDS and CMV infections were recruited at Peking Union Medical College Hospital from Jan 2010 to Aug 2017. Their clinical features and immune function were retrospectively analyzed. Patients with only HIV/AIDS in previous study were recruited as controls. Results: All 48 patients were at C3 stage, including 36 men and 12 women. Five of them were younger than 30 years old, 33 cases within 31-50 years old, and 10 cases older than 50 years old. Thirty-five patients had CD(4)(+)T lymphocytes ≤ 50 cells/µl, 7 cases with CD(4)(+)T cells 51-100/µl, 3 cases with 101-200 cells/µl, and 3 cases over 200 cells/µl. As to CMV infections, there were 31 cases of CMV viremia, 1 case of CMV encephalitis, 1 case of CMV enteritis, 5 cases of CMV pneumonia, and 9 cases of CMV retinitis. Other opportunistic infections were also common including 16 cases of pneumocystis pneumonia, 9 cases of tuberculosis, 5 cases of syphilis, 18 cases of digestive tract fungal infections, 8 cases of pulmonary fungal infections, 2 cases of EB virus infections, 2 cases of HIV encephalopathy/progressive multifocal leukoencephalopathy (PML), 3 cases of cryptococcal meningitis, 1 case of toxoplasma infection. In group of both CMV and HIV/AIDS infections, 100% patients had inverted CD(4)(+)/CD(8)(+) ratio. The immune activation marker CD(8)(+)CD(38)(+)/CD(8)(+) was higher (61.6%-98.8%) with a median value of 91.2% in 40 patients. HLA-DR(+)CD(8)(+)/CD(8)(+), another marker for T cell activation, was 25.5%-98.0% in 44 patients with a median value of 60.3%. Thirty-six patients had both immune activation markers positive. There was no significant difference in counts of B cells, natural killer cells, CD(4)(+) T cells, CD(8)(+) T cells and immune activation subsets stratified by gender and age (P>0.05). Meanwhile, neither serum HIV viral load nor serum CMV viral load was correlated with HLA-DR(+)CD(8)(+)/CD(8)(+), CD(8)(+)CD(38)(+)/CD(8)(+), CD(4)(+)T cell counts, and CD(4)(+)/CD(8)(+) ratio in the CMV and HIV/AIDS co-infection group (all P>0.05), while HIV viral load in HIV/AIDS only group was significantly correlated with HLA-DR(+)CD(8)(+)T/CD(8)(+), CD(38)(+)CD(8)(+)/CD(8)(+), CD(4)(+) T cell counts, CD(4)(+)/CD(8)(+) ratio (r=0.473, 0.575, -0.767 and -0.678, respectively, all P<0.05). Conclusions: CMV infections develop in HIV patients with advanced stage. CMV infection can cause life-threatening multiple organ lesions, especially in those with CD(4)(+) T cells less than 100 cells/µl. It is of great importance to screen CMV-IgM, pp65 antigen, CMV DNA to make early diagnosis and treatment.

Effects of laparoscopic splenectomy and esophagogastric devascularization on stress and immune function in patients with cirrhotic portal hypertension / 中华普通外科杂志

Objective To evaluate the effects of laparoscopic splenectomy and esophagogastric devascularization(LSPD) vs.open procedure(OSPD) on stress and immune function in patients with portal hypertension due to cirrhosis.Methods From June 2015 to June 2017 a total of 66 patients underwent LSPD or OSPD procedures.Results Plasma cortisol concentration in the LSPD group was lower than that in the OSPD group (Fgroup =18.85,P =0.020).Cortisol concentration in the two groups increased firstly and then decreased with time extension (Ftime =532.08,P =0.000).The level of CD3 +,CD4 +,CD4 +/CD8 + in LSPD group was higher than that in OSPD group,and the level of CD8 + was lower than that in OSPD group (Fgroup =3.55,21.47,154.84,64.29,P ＜ 0.05),the levels of CD3 +,CD4 + and CD4 +/CD8 + in the two groups first decreased and then increased with the extension of time,and the levels of CD8 + increased firstly and then decreased (Ftime =199.22,298.48,864.33,510.23,P ＜ 0.05),the increase range of CD3 +,CD4 +,and CD4 +/CD8 + in LSED group was higher than that in OSPD group (Finteraction =19.27,18.21,79.55,35.21,P ＜ 0.05);there was no statistical significance in the complications such as:rebleeding,thrombosis and splenic fever in the two groups (x2 =0.05,0.67,0.07,0.16,P=0.829,0.413,0.789,0.693).But the postoperative chest/cavity effusion and fever (＞ 38.0 ℃,and ＞ 3 days) is OSPD group is higher than in the LSPD group(x2 =5.49,6.68,P =0.019,0.010).Conclusion LSPD effectively reduces postoperative stress,protects immune function,decreases postoperative pleural and abdominal effusion and fever.

Clinical characteristics and T-lymphocyte subsets in 48 acquired immune deficiency syndrome patients with cytomegalovirus infections / 中华内科杂志

Objective To investigate the clinical features and T lymphocytes subsets in patients with acquired immune deficiency syndrome (AIDS) and cytomegalovirus (CMV) infection.Methods A total of 48 hospitalized patients with human immunodeficiency virus (HIV)-1/AIDS and CMV infections were recruited at Peking Union Medical College Hospital from Jan 2010 to Aug 2017.Their clinical features and immune function were retrospectively analyzed.Patients with only HIV/AIDS in previous study were recruited as controls,Results All 48 patients were at C3 stage,including 36 men and 12 women.Five of them were younger than 30 years old,33 cases within 31-50 years old,and 10 cases older than 50 years old.Thirty-five patients had CD4+T lymphocytes ≤ 50 cells/μl,7 cases with CD4+T cells 51-100/μl,3 cases with 101-200 cells/μl,and 3 cases over 200 cells/μl.As to CMV infections,there were 31 cases of CMV viremia,1 case of CMV encephalitis,1 case of CMV enteritis,5 cases of CMV pneumonia,and 9 cases of CMV retinitis.Other opportunistic infections were also common including 16 cases of pneumocystis pneumonia,9 cases of tuberculosis,5 cases of syphilis,18 cases of digestive tract fungal infections,8 cases of pulmonary fungal infections,2 cases of EB virus infections,2 cases of HIV encephalopathy/progressive multifocal leukoencephalopathy (PML),3 cases of cryptococcal meningitis,1 case of toxoplasma infection.In group of both CMV and HIV/AIDS infections,100％ patients had inverted CD4+/CD8+ ratio.The immune activation marker CD8+CD38+/CD8+ was higher (61.6％-98.8％) with a median value of 91.2％ in 40 patients.HLA-DR+ CD8+/CD8+,another marker for T cell activation,was 25.5％-98.0％ in 44 patients with a median value of 60.3％.Thirty-six patients had both immune activation markers positive.There was no significant difference in counts of B cells,natural killer cells,CD4+ T cells,CD8+ T cells and immune activation subsets stratified by gender and age (P＞0.05).Meanwhile,neither serum HIV viral load nor serum CMV viral load was correlated with HLA-DR+CD8+/CD8+,CD8+CD38+/CD8+,CD4+T cell counts,and CD4+/CD8+ ratio in the CMV and HIV/AIDS co-infection group(all P＞0.05),while HIV viral load in HIV/AIDS only group was significantly correlated with HLA-DR+CD8+T/CD8+,CD38+CD8+/CD8+,CD4+ T cell counts,CD4+/CD8+ ratio (r=0.473,0.575,-0.767 and-0.678,respectively,all P＜0.05).Conclusions CMV infections develop in HIV patients with advanced stage.CMV infection can cause life-threatening multiple organ lesions,especially in those with CD4+ T cells less than 100 cells/μl.It is of great importance to screen CMV-IgM,pp65 antigen,CMV DNA to make early diagnosis and treatment.

Impact of Aging on the Frequency, Phenotype, and Function of CD161-Expressing T Cells.

Immune-aging is associated with perturbed immune responses in the elderly. CD161-expressing T cells, i.e., the previously described subsets of CD161+ CD4+ T cells, CD161high CD8+ T cells, and CD161int CD8+ T cells, are highly functional, pro-inflammatory T cells. These CD161-expressing T cells are critical in immunity against microbes, while possibly contributing to autoimmune diseases. So far, little is known about the impact of aging on the frequency, phenotype, and function of these CD161-expressing T cells. In the current study, we investigated the impact of aging on CD161+ CD4+ T cells, CD161high CD8+ T cells, and CD161int CD8+ T cells in peripheral blood samples of 96 healthy subjects (age 20-84). Frequencies of CD161+ CD4+ T cells and CD161int CD8+ T cells were stable with aging, whereas frequencies of CD161high CD8+ T cells declined. Although CD161high CD8+ T cells were mostly T cell receptor-Vα7.2+ mucosal-associated invariant T cells, CD161 expressing CD4+ and CD8+ T cells showed a limited expression of markers for gamma-delta T cells or invariant natural killer (NK) T cells, in both young and old subjects. In essence, CD161-expressing T cells showed a similar memory phenotype in young and old subjects. The expression of the inhibitory NK receptor KLRG1 was decreased on CD161+ CD4+ T cells of old subjects, whereas the expression of other NK receptors by CD161-expressing T cells was unaltered with age. The expression of cytotoxic effector molecules was similar in CD161high and CD161int CD8+ T cells of young and old subjects. The ability to produce pro-inflammatory cytokines was preserved in CD161high and CD161int CD8+ T cells of old subjects. However, the percentages of IFN-γ+ and interleukin-17+ cells were significantly lower in CD161+ CD4+ T cells of old individuals than those of young individuals. In addition, aging was associated with a decrease of nonclassic T helper 1 cells, as indicated by decreased percentages of CD161-expressing cells within the IFN-γ+ CD4+ T cell compartment of old subjects. Taken together, aging is associated with a numerical decline of circulating CD161high CD8+ T cells, as well as a decreased production of pro-inflammatory cytokines by CD161+ CD4+ T cells. These aging-associated changes could contribute to perturbed immunity in the elderly.

Immunophenotyping of circulating mononuclear cells in active pulmonary tuberculosis.

INTRODUCTION: Interpreting the interactions between M. tuberculosis and the host innate and adaptive immune defense mechanisms is mandatory for understanding the pathogenesis of active pulmonary TB (APTB). The aim was to describe the distribution of mononuclear cells in APTB and their relation to disease severity. METHODOLOGY: A case-control study of peripheral blood CD4+ T cells, CD8+ T cells, B-lymphocytes, NK cells, T regulatory lymphocytes (Tregs) and monocytes by flow cytometry. The patients had clinical presentations of APTB, positive tuberculin skin tests, acid-fast bacilli smears and sputum cultures using BACTEC 960. RESULTS: There was a significant decrease in the haemoglobin level and the absolute lymphocytic count (p < 0.01), while both the neutrophil count and erythrocyte sedimentation rate showed significant increase in the APTB patients compared to HC with p-values < 0.001 and < 0.0001 respectively. Both the CD4+/CD8+ ratio and the percentages of CD3-CD19+ cells were significantly lower in APTB patients (p = 0.03 and p = 0.005 respectively). The percentages of CD4+, CD8+, CD3-CD19+, CD14+, and CD3-CD (16+56)+ cells showed no significant differences, when comparing either disease severity groups, or cavitated and non-cavitated groups of APTB patients. There was significant increase in the CD4+25+ lymphocytes in the advanced APTB patients than in the mild disease group (p < 0.05). CONCLUSIONS: B-lymphocytes and CD4/CD8 ratios were significantly lower in the APTB patients than controls with no association with disease severity. CD4+ CD25+hi Tregs were significantly higher in the advanced versus mild groups.

Study on the correlation of CD8 +T cell with CD8low T cell subsets and CD8high T cell subsets / 中华检验医学杂志

Objective To investigate the relationship between the CD 8 +T cells results of clinical automatic analysis platform and the CD8lowT and CD8highT cell subsets.Methods A total of 1316 cases of lymphocyte and flow cytometry data were collected from the First Affiliated Hospital of China Medical University from December 2015 to September 2016 by cross-sectional study. There were 287 cases of malignant tumor , 389 cases of autoimmune disease , 320 healthy people and 320 cases of HIV infection , then to get automatic analysis platform returns result of CD 8+T cell.FlowJo software was used to analyze the CD8low T and CD8high T lymphocyte subsets in the patients , and the results were compared with the results of CD8 +T cells returned by the clinical automatic analysis platform .Results The results of clinical returns CD8 +T cells were consistent with the results of CD 8high T cells in patients with different diseases , and were not exactly the same as the results of CD8lowT cells, and the difference was as follows:the results of CD8low T cells in HIV-infected patients were significantly lower than those of healthy people (56.2 ±42.0, 68.8 ± 45.9, cells/μl P<0.001), which were different from the clinical results of CD 8 +T cells.The results of clinical report of CD8 +T cells were statistically correlated with CD8high T cells and CD8low T cells, and the correlation between CD8 +T cells and CD8highT were higher than that of CD8lowT cells.There was a positive correlation between CD8low T cell count and CD4 +T cell count ( r=0.204, P<0.001) .CD8low T was significantly higher in patients on antiviral treatment than that in untreated group (58.3 ±43.9, 42.9 ± 26.5, cells/μl, P<0.001).After treatment for more than 2 years, the CD8lowT cells in patients with CD4<500 cells/μl were significantly lower than those in patients with CD 4>500 cells/μl (50.1 ±47.0, 66.3 ±46.6, cells/μl, P<0.001).Conclusions The clinical report of CD8 +T cells was consistent with the results of CD8highT cells, and there was a great difference between the results of CD 8lowT cells and the results of CD8 +T cells.CD8low T cells were significantly reduced in HIV infected patients , and CD8low T cells could be effectively reconstructed by antiviral therapy .

Specific immune response in neonate Holstein heifer calves fed fresh or frozen colostrum / Resposta imune específica em bezerras holandesas recém-nascidas alimentadas com colostro fresco ou congelado

The objective of this study was to evaluate the influence of viable cells from colostrum on immune development in dairy heifer calves during the first 28 days of life. The animals were distributed between 2 groups: COL+ (n=9) receiving fresh whole colostrum from their own damns; and COL- (n=10) receiving pooled frozen colostrum, containing no viable cells, from a pool of donor cows. These calves were assessed before colostrum intake (D0), 48 hours of age (D2), and weekly from D7 to D28. The development of immunity was evaluated by assessment of the phenotype of blood leukocyte subsets, and induced cytokine production after 72 hours of stimulation in culture with concanavalin A (ConA), killed Staphylococcus aureus (S.aureus) and killed Escherichia coli (E. coli) by peripheral blood mononuclear cell (PBMC). The clinical history of these calves was marked by a high frequency of diarrhea in both groups. However, COL- had greater diarrhea intensity scores (fecal score~3 of 4), and rectal temperature on D7 than COL+ calves. Moreover, bronchopneumonia (n=1) and navel inflammation were observed only in COL- calves. COL- had a lower concentration of serum iron, and a higher absolute number of lymphocytes on D7 than COL+. COL- also had a higher percentage of anemic calves than the COL+ calves on D21 and D28. In general, the percent of cells within each subset of leukocytes was similar between the groups over the experiment, except on week 1 when COL- calves had a higher percentage of lymphocytes expressing CD45RO+ (P=0.07). A steady increase in CD45RO+ and concomitant decline in CD45RO- leukocytes was observed over the course of the study, indicating the development of immune memory. The proportion of CD14MHCII+ leukocytes increased with age (P≤0.05). The median background cytokine production by PBMC that were not stimulated was below the level of detection of the assays used for both groups. The PBMC from COL+ calves stimulated with ConA secreted a larger quantity of IL-17 week 2 (COL+=2060.0pg/mL and COL-=0.0pg/mL, P=0.00). PBMC from COL+ calves stimulated with killed S. aureus whole cell antigen (P=0.05) and killed E. coli whole cell antigen (P=0.05) also secreted higher levels of IL17 than COL- calves at week 4. Clear production of IL17 was observed in PBML from COL+ calves at week 2, but the difference was not statistical different between groups. In conclusion, calves fed fresh and frozen colostrum showed no difference in cells subset profile overall. The increased percentage of leukocytes expressing the memory CD45RO+ or CD14MHCII+ over the course of the experiment indicated a maturation of the adaptive immune response after natural exposure to pathogens in the environment of the calf. The enhanced IL17 secretion by COL+ calves indicated that viable maternal cells modulated T-cell Th17 production that was primed by bacterial antigens. This mechanism could be responsible for quick and efficient activation of neutrophils for bacterial clearance. The differences in cytokine production observed between groups may help to explain the different clinical pictures observed for calves COL+ and COL- calves.(AU)

O objetivo desta pesquisa foi avaliar a influência das células do colostro no desenvolvimento imune em bezerras leiteiras durante os primeiros 28 dias de vida. Os animais foram distribuídas em 2 grupos: COL+ (n=9) recebeu colostro fresco de suas próprias mães; e COL- (n=10) recebeu pool de colostro congelado sem células viáveis oriundo de vacas doadoras. Estas bezerras foram avaliadas antes da ingestão do colostro (D0); às 48 horas (D2) e semanalmente entre o D7 e D28. O desenvolvimento da imunidade foi avaliada pela fenotipagem das subpopulações celulares do sangue e produção de citocinas pelas células mononucleares sanguíneas após 72 horas de estimulação com concanavalina A (ConA), Staphylococcus aureus (S. aureus) e Escherichia coli (E. coli) inativadas. O histórico clínico das bezerras foi marcado por elevada frequência de diarreias em ambos os grupos. Entretanto, COL- apresentou maior intensidade de diarreia (escore de fezes ~ 3 de 4) e maior temperatura retal no D7 do que as bezerras COL+. Além disso, broncopneumonia (n=1) e inflamações umbilicais foram diagnosticadas apenas nas bezerras COL-. O grupo COL- apresentou menor concentração de ferro sérico e maior número de linfócitos no D7 do que o grupo COL+. COL- também apresentou maior frequência de anemias que o grupo COL+ no D21 e D28. Em geral, a fração das subpopulações celulares foram semelhantes entre os dois grupos ao longo do tempo estudado, exceto na semana 1, onde as bezerras COL- apresentaram maior proporção de CD45RO+ (P=0.07). Observou-se um constante aumento de CD45RO+ com declínio concomitante de CD45RO- ao longo do estudo, indicando o desenvolvimento da resposta imune. A proporção de células CD14MHCII+ aumentou de acordo com a idade (P≤0.05). As medianas das citocinas produzidas a partir do PBMC não estimuladas apresentaram valores abaixo do nível de detecção em ambos os grupos. O PBMC do COL+ estimulado com ConA secretou elevada concentração de IL17 na semana 2 (COL+=2060.0pg/mL e COL-=0.0pg/ml, P=0.00). PBMC do COL+ estimuladas com S. aureus inativado (P=0.05) e E. coli inativada (P=0.05) secretaram níveis mais elevados de IL17 que as bezerras COL- na semana 4. Outros picos de IL17 foram observados no COL+ na semana 2, porém não foi possível detectar diferenças estatísticas entre os grupos. Conclui-se que as bezerras alimentadas com colostro fresco e congelado apresentaram perfil similar entre as proporções das subpopulações celulares. O aumento de células expressando os marcadores de memória CD45RO+ e CD14MHCII+ demonstram, ao longo do experimento, o amadurecimento do sistema imune específico das bezerras após estimulação natural por patógenos após o nascimento. A maior secreção de IL17 pelas células das bezerras COL+ indica que as células maternas podem modular resposta imune Th17 direcionada aos antígenos bacterianos. Este mecanismo poderia ser responsável pela rápida e eficiente quimiotaxia de neutrófilos e eliminação dos microrganismos bacterianos. Os diferentes perfis de citocinas podem ser responsáveis pelos diferentes históricos clínicos relatados para as bezerras COL+ e COL-.(AU)

Imunidade celular em vacas Holandesas soropositivas para o Vírus da Leucose Bovina (BLV) durante o período de transição / Cellular immunity in Holstein cows seropositive for Bovine Leukosis Virus (BLV) during the transition period

Esta pesquisa avaliou a dinâmica dos leucócitos e das subpopulações de linfócitos em vacas Holandesas soropositivas para o BLV no período de transição. Amostras de sangue (n=72) provenientes de 12 vacas foram coletadas entre as semanas -2 e +3 para a realização do leucograma, imunofenotipagem, dosagem de cortisol e haptoglobina (Hp). O perfil leucocitário foi caracterizado por leucocitose, neutrofilia, monocitose e eosinopenia próximo ao parto. Linfocitose e elevada proporção de linfócitos B CD21+ foram achados constantes entre as semanas -2 e +3; assim, as vacas foram testadas e confirmadas soropositivas para o BLV. Os valores das subpopulações de linfócitos T apresentaram-se baixos durante o período de transição, observando-se dois picos máximos que coincidiram com as elevações nas concentrações de cortisol no parto (2,11µg/dL) e semana +3 (1,97µg/dL). Hp apresentou aumento crescente de -2 (166µg/mL) a +3 (576µg/mL), provavelmente associada à elevada taxa de infecções uterinas observadas nas semanas +2 e +3. As vacas soropositivas para o BLV apresentaram leucograma de estresse próximo ao parto, exceto para linfócitos. A linfocitose e as elevadas proporções de células B CD21+, associadas com as baixas proporções de células T, podem ser indicativo de imunossupressão e predisposição aos processos inflamatórios no período pós-parto.(AU)

This research evaluated the dynamics of leukocytes and lymphocytes subsets in seropositive Holstein cows for BLV during the transition period. Blood samples (n=72) from 12 cows were harvested from week -2 up to week +3 to perform leukogram, immunophenotyping, cortisol and haptoglobin (Hp). Leukocytes pattern was characterized by leukocytosis, neutrophilia, monocytosis and eosinopenia around calving. Lymphocytosis and high proportions of B cells CD21+ were a constant finding between week -2 and +3, thus cows were tested and confirmed seropositive for BLV. The values of T lymphocytes subsets were low during the transition period, observing two peaks that coincided with high levels of cortisol at delivery (2.11µg/dL) and week +3 (1.97µg/dL). Hp had gradual increase from week -2 (166µg/mL) until week +3 (576g/mL) probably due to high rate of uterine infection detected between week +2 and +3. The seropositive cows for BLV presented stress leukogram around delivery, except for lymphocytes. Lymphocytosis and the high proportions of B cells, associated with the low proportions of T lymphocytes, can be indicative of immunosuppression and predisposition to the inflammatory process observed in the post-partum period.(AU)

Effects of Dijincao Tablets on T-lymphocytes subsets and cytokines in patients with immune thrombocytopenia / 中成药

AIM To observe the effects of Dijincao Tablets (Euphorbia humifusa Willd.) on T-lymphocytes subsets and cytokines in patients with immune thrombocytopenia (ITP).METHODS Fifty-six patients who were first diagnosed ITP at hematology department of our hospital from Jan.2014 to Dec.2015 were divided into observation group (twenty-eight cases) and control group (twenty-eight cases) by Stratified blocked randomization.The patients in the observation group were given Dijincao Tablets (4.8 g,tid) based on the conventional treatment,while the patients in the control group only accepted conventional treatment according to guidelines.Adverse events were collected during the treatment.Fasting venous blood was obtained from all patients in the morning on the second day of admission and the weekend of the forth week (30th day after admission) to detect observation indexes,and then the comprehensive effects were evaluated.The levels of T-lymphocytes subsets of CD3 +,CD4 +,CD8 + and CD4 +/CD8 + were examined by flow cytometry,and serum cytokine levels of IL-2,IFN-γ,IL-4 and IL-10 were determined by ELISA.RESULTS The total effective rate was 92.9％ in the observation group,which was higher than 82.1％ in the control group.The platelet level in the observation group was significantly higher than that in the control group.Compared with the control group,the levels of CD3 +,CD4 + and CD4 +/CD8 + in the observation group were significantly increased,the levels of IL-2 and IFN-γwere significantly decreased,and the ratio of IFN-γ/IL-4 was significantly decreased.CONCLUSION Dijincao Tablets can correct and regulate the disorder and ratio of T-lymphocyte subsets,and decrease the ratio of IFN-γ/IL-4,then achieve therapeutic effect on patients with immune thrombocytopenia.

The changes of immunin function in pediatric patients with severe sepsis / 中国小儿急救医学

Objective To investigate the association of immunological indicators with the severity and prognosis of pediatric patients with severe sepsis.Methods We enrolled 82 pediatric patients with severe sepsis admitted to pediatric intensive care unit (PICU) at Shanghai Children′s Hospital between March 2013 and February 2017 as septic group.Fifteen healthy children served as control group.The blood samples were collected within 24 hours after admission and on day 7 after treatment.The levels of immunoglobulin (IgG,IgM and IgA) were analyzed by automatic special protein analyzer,and the proportion of T-lymphocyte subgroup (CD3+,CD4+,CD8+ and CD19+) and natural killer (NK) cells (CD16+ and CD56+) in peripheral blood were detected by flow-cytometry.Results The levels of IgG,IgM and IgA had no statistical differences between septic group and control group(P>0.05).Interestingly,the proportion of NK cells in pediatric patients with severe sepsis was significantly lower compared to the control group,and the number of NK cells was significantly increased after 7 days treatment compared with that within 24 hours after admission[(3.7±1.9)% vs.(11.5±1.9)%,P<0.05].In addition,the proportions of T-lymphocyte subgroups including CD3+,CD4+ and CD8+ were significantly decreased in patients of septic group compared with control group[(62.8±8.5)% vs.(70.9±2.3)%,(33.3±7.0)% vs.(39.8±1.8)% and(22.6±2.8)% vs.(34.8±15.6)%,respectively,all P<0.05].Moreover,the proportions of NK cells,CD3+ and CD4+ T lymphocytes subsets in peripheral blood of patients with severe sepsis were positively associated with pediatric critical illness score(P<0.05),and negatively associated with pediatric risk of mortality score Ⅲ and the number of dysfunction organs(all P<0.05).Furthermore,the proportions of NK cells and CD3+ and CD4+ T lymphocytes in peripheral blood of non-survivor with severe sepsis were significantly lower than those in the survivor[(1.5±0.5)% vs.(4.7±1.4)%,(55.1±5.0)% vs.(66.4±7.4)%,(29.7±5.2)% vs.(35.0±7.2),P< 0.05].Conclusion The proportion of NK cells and CD3+ and CD4+ T lymphocytes subsets in peripheral blood decreases in pediatric patients with severe sepsis,which is associated with severity and prognosis of severe sepsis.

Immune cell changes in response to a swimming training session during a 24-h recovery period.

Understanding the impact of training sessions on the immune response is crucial for the adequate periodization of training, to prevent both a negative influence on health and a performance impairment of the athlete. This study evaluated acute systemic immune cell changes in response to an actual swimming session, during a 24-h recovery period, controlling for sex, menstrual cycle phases, maturity, and age group. Competitive swimmers (30 females, 15 ± 1.3 years old; and 35 males, 16.5 ± 2.1 years old) performed a high-intensity training session. Blood samples were collected before, immediately after, 2 h after, and 24 h after exercise. Standard procedures for the assessment of leukogram by automated counting (Coulter LH 750, Beckman) and lymphocytes subsets by flow cytometry (FACS Calibur BD, Biosciences) were used. Subjects were grouped according to competitive age groups and pubertal Tanner stages. Menstrual cycle phase was monitored. The training session induced neutrophilia, lymphopenia, and a low eosinophil count, lasting for at least 2 h, independent of sex and maturity. At 24 h postexercise, the acquired immunity of juniors (15-17 years old), expressed by total lymphocytes and total T lymphocytes (CD3(+)), was not fully recovered. This should be accounted for when planning a weekly training program. The observed lymphopenia suggests a lower immune surveillance at the end of the session that may depress the immunity of athletes, highlighting the need for extra care when athletes are exposed to aggressive environmental agents such as swimming pools.

Efficacy Observation ofFufang Shenlu Granule for Kidney-yang Deficiency Aplastic Anemia and Its Effects on CD4+ T Lymphocyte Subsets / 中国中医药信息杂志

Objective To observe the clinical efficacy ofFufang Shenlu Granule for kidney-yang deficiency aplastic anemia (AA) and its effects on CD4+ T lymphocyte subsets, and explore its mechanism.Methods Forty AA patients were randomly divided into treatment group and control group, 20 cases in each group. The treatment group was treated withFufang Shenlu Granule combined with conventional western medicine therapy, while the control group was only treated with conventional western medicine therapy. The treatment lasted for 6 months, and follow-up visits lasted for at least one year. The clinical efficacy and T lymphocyte subsets ratio changes before and after treatment in the two groups were observed.Results The clinical efficacy of treatment group was better than that of control group (P<0.05). After treatment, the blood cell counts of the two groups were improved than those of before treatment (P<0.05,P<0.01). After treatment, the red blood cell count and hemoglobin of treatment group were higher than those of control group (P<0.05). Compared with the data before treatment in treatment group, the ratio of CD4+CD25+ cells in all CD4+ cells, and the ratio of CD4+CD25+FoxP3+ cells in all CD4+CD25+ cells increased (P<0.05); the ratio of Th1 and Th2 cells in all CD3+CD4+ decreased (P<0.05). The ratio of Th2 cells was lower than that before treatment in the control group (P<0.05).ConclusionFufang Shenlu Granule can enhance the treatment efficiency of AA with kidney-yang deficiency syndrome. The mechanism may be related to the decrease of Th1 cells, the increase of CD4+CD25+Tregs and their FoxP3 expression.

Analysis difference of lymphocyte subgroups between children with severe pneumonia and non-severe pneumonia / 中国免疫学杂志

Objective:To observe the levels of lymphocyte subgroups in children with severe pneumonia and non -severe pneumonia , and to investigate the change of celluer immune funciton of patients , moreover provide available reference for treatment.Methods:Flow cytometry instrument was used to detect peripheral blood CD 3+T, CD3+CD4+T, CD3+CD8+T, CD19+B lymphocytes and NK cells in 27 children with severe pneumonia and 28 children with non-severe pneumonia.In addition 20 heathy children were selected as the controls .Results:Compared with the heathy control group ,the levels of CD3+T,CD3+CD4+T,CD3+CD8+T lymphocytes and NK cells in children with severe pneumonia decreased significantly (P<0.01,P<0.05,P<0.01,P<0.05),while CD4/CD8 ratio,CD19+B cells were significantly increased (P<0.05,P<0.01).The level of CD3+CD8+T lymphocytes in children with non-severe pneumonia significantly lower than the heathy control group (P<0.01),but the level of CD19+B cells in children with non-severe pneumonia significantly higher than the heathy control group (P<0.01).Compared with non-severe pneumonia group.The levels of CD3+and CD3+CD8+T lymphocytes in children with severe pneumonia decreased significantly (P<0.01),however CD19+B cells were significantly increased ( P<0.01 ) .Conclusion: Immune dysfunction exists in both of children with severe pneumonia and non-severe pneumonia.Children with severe pneumonia has more serious damage in inmmunity .This provides certain theoretical basis for clinical e-valuation and treatment .