Clinical features, immunologic parameter and treatment outcome of Chinese tuberculosis patients with or without DM.

Background: The coexistence of diabetes mellitus (DM) and pulmonary tuberculosis (PTB) poses a significant health concern globally, with their convergence presenting a considerable challenge to healthcare systems. Previous research has highlighted that comorbidities can mutually influence and exacerbate immune disorders. However, there is a paucity of data on the impact of DM on immunological features and treatment responses in the TB population in China. Methods: From January 2020 to June 2022, 264 cases of pulmonary tuberculosis patients (82 DM patients and 182 non-DM patients) hospitalized in our center were selected. 80 patients with TB with DM (TB-DM) and 80 patients with TB without DM (TB-NDM) were enrolled into the final analysis by propensity score matching for age, gender and involved lung field at a ratio of 1:1. The clinical characteristics, immunological features and treatment response were compared between the two groups. Results: After propensity score matching, no differences in the general features such as age gender, involved lung field, the incidence of retreatment and WBC count were found between the two groups. Compared to TB-NDM group, the TB-DM group exhibited a higher positive rate of sputum smear and incidence of cavitary lesions. Immunological features analysis revealed that the TB-DM patients had higher levels of TNF-α [pg/ml; 8.56 (7.08-13.35) vs. 7.64 (6.38-10.14) p = 0.033] and IL-8 [pg/ml; 25.85 (11.63-58.40) vs. 17.56 (6.44-39.08) p = 0.003] but lower CD8+ T lymphocyte count [cells/mm3; 334.02 (249.35-420.71) VS 380.95 (291.73-471.25) p = 0.038]. However, there was no significant difference in serum IL-6 concentration and CD4+ T lymphocyte count between the two groups. After 2 months of anti-tuberculosis treatment, 39 (24.4%) cases had suboptimal treatment response, including 23 (28.7%) TB-DM patients and 16 (20%) TB-NDM patients. There was no difference in suboptimal response rate (SRR) was found between the two groups (p = 0.269). The multivariate logistic regression analysis indicated that retreatment for TB [AOR: 5.68 (95%CI: 2.01-16.08), p = 0.001], sputum smear positivity [AOR: 8.01 (95%CI: 2.62-24.50), p = 0.001] were associated with SRR in all participants, and in TB-DM group, only sputum smear positivity [AOR: 16.47 (1.75-155.12), p = 0.014] was positive with SRR. Conclusion: DM is a risk factor for pulmonary cavity formation and sputum smear positivity in TB population. Additionally, TB-DM patients is characterized by enhanced cytokine responses and decreased CD8+ T lymphocytes. The retreatment for TB and sputum smear positivity were associated with the occurrence of suboptimal treatment response.

The role of immune and inflammatory-related indicators in cognitive dysfunction and disease severity in patients with parkinson's disease.

We aimed to explore the role of immune and inflammatory indicators in cognitive dysfunction and disease severity in patients with Parkinson's disease (PD). A total of 123 patients with Parkinson's disease were enrolled in the PD group and 49 healthy volunteers in the control group. The patients with PD were further divided into 2 subgroups by evaluating cognitive function using the Montreal Cognitive Assessment (MoCA) and Mini-Mental State Examination (MMSE): the normal cognitive function (PD-NCI) group and the mild cognitive impairment (PD-MCI) group. Moreover, the PD patients were also divided into 2 subgroups using the defined scale of the Hoehn and Yahr (H-Y) stage: the early-stage group and the middle- and late-stage group. Immune and inflammatory indicators, including serum Aß1-42, Tau, CD4+, CD8+, CD3+, B lymphocytes cell, NK cell, Th17 cell, Treg cell, IL-6, IL-17, and TNF-α levels, were evaluated and analyzed to explore the potential correlation with the cognitive dysfunction and disease severity of PD. Among the 123 PD patients, 60 (48.8%) were diagnosed with mild cognitive impairment. Aß1-42, CD4+, CD8+, CD3+, and Treg levels observed in the PD-NCI group were lower than the control group (P < 0.001), while higher than the PD-MCI group (P < 0.001). The levels of Tau, Th17, IL-6, IL-17, and TNF-α observed in the PD-NCI group were higher than the control group (P < 0.001), while lower than in the PD-MCI group (P < 0.01). Using the same method, the results of the early-stage group and the middle- and the late-stage group were the same as above. Logistic regression analysis and ROC curve estimation were performed and indicated that the variation of Tau, CD8+, Treg, TNF-α levels was associated with cognitive decline in PD patients, and may serve as markers of PD onset. Furthermore, the variation of Aß1-42, IL-6, and TNF-α levels was found to correlate with the disease severity of PD. The immune and inflammatory-related indicators may represent an important factor in the pathogenesis of PD, cognitive dysfunction, and disease severity. The variation of Tau protein, CD8+, Treg, and TNF-α levels are associated with the cognitive dysfunction of PD, which may be considered as onset markers. Moreover, the variation of Aß1-42, IL-6, and TNF-α levels can predict the progression of PD.

[Clinical study of 19 cases of steroid-refractory gastrointestinal acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation with fecal microbiota transplantation].

Objective: To investigate the clinical efficacy of fecal microbiota transplantation (FMT) for treating steroid-refractory gastrointestinal acute graft-versus-host disease (GI-aGVHD) . Methods: This analysis included 29 patients with hematology who developed steroid-refractory GI-aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Huaian Hospital Affiliated to Xuzhou Medical University from March 2017 to March 2022. Among them, 19 patients underwent FMT treatment (the FMT group) and 10 patients did not (the control group). The efficacy and safety of FMT were assessed, as well as the changes in intestinal microbiota abundance, lymphocyte subpopulation ratio, peripheral blood inflammatory cytokines, and GVHD biomarkers before and after FMT treatment. Results: ① Complete remission of clinical symptoms after FMT was achieved by 13 (68.4%) patients and 2 (20.0%) controls, with a statistically significant difference (P<0.05). Intestinal microbiota diversity increased and gradually recovered to normal levels after FMT and FMT-related infections did not occur. ②The proportion of CD3(+) and CD8(+) cells in the FMT group after treatment decreased compared with the control group, and the ratio of CD4(+), regulatory T cells (Treg), and CD4(+)/CD8(+) cells increased (all P< 0.05). The interleukin (IL) -6 concentration in the FMT group was lower than that in the control group [4.15 (1.91-5.71) ng/L vs 6.82 (2.40-8.91) ng/L, P=0.040], and the IL-10 concentration in the FMT group was higher than that in the control group [12.11 (5.69-20.36) ng/L vs 7.51 (4.10-9.58) ng/L, P=0.024]. Islet-derived protein 3α (REG3α) was significantly increased in patients with GI-aGVHD, and the REG3α level in the FMT group was lower than that in the control group after treatment [30.70 (10.50-105.00) µg/L vs 74.35 (33.50-139.50) µg/L, P=0.021]. Conclusion: FMT is a safe and effective method for the treatment of steroid-refractory GI-aGVHD by restoring intestinal microbiota diversity, regulating inflammatory cytokines, and upregulating Treg cells.

[Analysis on Cellular Immune Function Recovery after HLA Mismatched Allogeneic Hematopoietic Stem Cell Micro-Transp- lantation in Elderly AML Patients].

OBJECTIVE: To investigate the recovery of cellular immunity in elderly patients with acute myeloid leukemia (AML) after micro-transplantation (MST) and the changes of cellular immunity during relapse, as well as their clinical significance. METHODS: A total of 41 elderly AML patients who received MST treatment in a single center and 25 healthy elderly people were included. Immune function among different age groups in normal population was compared. Furthermore, immune fuction was compared between elderly AML patients of different age groups who achieved continuous complete remission (CR) after MST treatment and normal controls, between high risk group and medium-low risk group, as well as among before diagnosis, after CR, and relapse. Peripheral blood of patients and normal controls was collected, and the percentage of lymphocyte subsets was detected by multi-color flow cytometry. RESULTS: Thirty-five patients achieved CR after MST treatment while six patients did not. After MST treatment, CD3+ T cells, CD8+T cells and activated T cells in all age groups were higher than normal. Significant recovery of CD3+ and CD8+T cells was observed in both high risk and medium-low risk groups, and the overall recovery of immune cells in medium-low risk group was better. It was also observed that B lymphocytes and NK cells could not return to normal levels within 1 year after MST treatment. The proportion of CD3+ T cells, CD4+ T cells, and CD4/CD8 ratio were significantly decreased during relapse compared with continuous CR after MST (P<0.05). CONCLUSION: MST treatment can promote the recovery of CD3+T cells, CD8+T cells and other killer cells, so as to improve the cellular immune function of elderly patients, which provides a new immune cell therapy for elderly AML.

Predictors of COVID-19 Severity in Elderly Patients Infected by Omicron in China, 18 December 2022-5 February 2023.

Purpose: To analyze the clinical characteristics and prognosis of patients hospitalized with non-severe, severe pneumonia and death in Omicron COVID-19. Patients and Methods: We collected clinical data from 118 patients with COVID-19 in China from 18 December, 2022 and 5 February, 2023. According to the outcome, the patients were divided into non-severe group, severe group and death group. Subsequently, we statistically analyzed the general condition, clinical manifestations, laboratory parameters, NLR, MLR, PLR and HALP of these groups. We also retrospectively analyzed the possible factors affecting the prognostic regression of patients with COVID-19. Results: A total of 118 COVID-19 patients were enrolled in this study, including 64 non-severe patients, 38 severe patients and 16 death patients. Compared with the non-severe group, T lymphocytes, B lymphocytes, Th1, Th2, Th17, Treg cells, IgA, IgG, IgM in the severe and death groups decreased more significantly (P<0.05). The levels of myocardial markers, ALT, AST, BUN, Cr, D-dimer, fibrinogen, NLR, MLR and PLR in the severe and death groups were significantly higher than those in the non-severe group (P<0.05). The level of HALP was significantly lower than that of non-severe group (P<0.05). MLR is not only an independent risk factor for the transition from non-severe to severe disease, but also an independent risk factor for predicting the possibility of death in COVID-19 patients. Conclusion: The analysis of COVID-19 patients in China showed that severe patients were older, more likely to have related complications, lower lymphocyte count, liver and kidney function disorder, glucose and lipid metabolism disorders, myocardial injury, and abnormal coagulation function, suggesting the need for early anticoagulant therapy. In addition, NLR, MLR, PLR and HALP can be used as biomarkers to evaluate the severity and prognosis of COVID-19 patients.

Predictive value of circulating tumor cell counts during the treatment of cancer: interactions with the blood microenvironment.

OBJECTIVE: This study aimed to evaluate the prognostic value of circulating tumor cell (CTC) in tumor patients during treatment. METHODS: This study retrospectively analyzed clinical data obtained from 174 cancer patients during treatment. The relationship between the CTC counts and clinicopathological variables was analyzed. A ROC curve was applied to determine the optimal cut-off values and assess the predictive ability of the prognostic indicators. The overall survival (OS) for different prognostic factors was calculated using the Kaplan-Meier method, and the difference between the survival curves was then compared using the log-rank test. Cox regression model was used to investigate the effect of independent factors on patients' survival. RESULTS: The CTC-positive rate was positively correlated with the clinicopathological variables of TNM stage, tumor differentiation, serum CEA level, and ki-67%. In the differential analysis of hematological microenvironment parameters in CTC-positive and CTC-negative samples, the complete blood count, blood biological chemistry, tumor markers (CEA, CA19-9, CA72-4), and lymphocyte subpopulation were statistically significant. The results of the ROC curve analysis indicated that the serum CEA level was the best diagnostic indicator to discriminate the CTC count in tumor patients. Additionally, the results of the univariate and multivariate analyses of OS in relation to clinical variables revealed that the CTC counts were an independent prognostic factor for unfavorable OS. CONCLUSION: The CTC counts in patients with tumors undergoing treatment were significantly correlated with hematological microenvironment parameters. The detection of CTCs may therefore be used as an indicator of tumor prognosis.

Clinical study of 19 cases of steroid-refractory gastrointestinal acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation with fecal microbiota transplantation / 中华血液学杂志

Objective: To investigate the clinical efficacy of fecal microbiota transplantation (FMT) for treating steroid-refractory gastrointestinal acute graft-versus-host disease (GI-aGVHD) . Methods: This analysis included 29 patients with hematology who developed steroid-refractory GI-aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Huaian Hospital Affiliated to Xuzhou Medical University from March 2017 to March 2022. Among them, 19 patients underwent FMT treatment (the FMT group) and 10 patients did not (the control group). The efficacy and safety of FMT were assessed, as well as the changes in intestinal microbiota abundance, lymphocyte subpopulation ratio, peripheral blood inflammatory cytokines, and GVHD biomarkers before and after FMT treatment. Results: ① Complete remission of clinical symptoms after FMT was achieved by 13 (68.4%) patients and 2 (20.0%) controls, with a statistically significant difference (P<0.05). Intestinal microbiota diversity increased and gradually recovered to normal levels after FMT and FMT-related infections did not occur. ②The proportion of CD3(+) and CD8(+) cells in the FMT group after treatment decreased compared with the control group, and the ratio of CD4(+), regulatory T cells (Treg), and CD4(+)/CD8(+) cells increased (all P< 0.05). The interleukin (IL) -6 concentration in the FMT group was lower than that in the control group [4.15 (1.91-5.71) ng/L vs 6.82 (2.40-8.91) ng/L, P=0.040], and the IL-10 concentration in the FMT group was higher than that in the control group [12.11 (5.69-20.36) ng/L vs 7.51 (4.10-9.58) ng/L, P=0.024]. Islet-derived protein 3α (REG3α) was significantly increased in patients with GI-aGVHD, and the REG3α level in the FMT group was lower than that in the control group after treatment [30.70 (10.50-105.00) μg/L vs 74.35 (33.50-139.50) μg/L, P=0.021]. Conclusion: FMT is a safe and effective method for the treatment of steroid-refractory GI-aGVHD by restoring intestinal microbiota diversity, regulating inflammatory cytokines, and upregulating Treg cells.

Sleep quality and influencing factors and correlation with T-lymphocyte subpopulation counts in patients with pulmonary tuberculosis: a cross-sectional study.

BACKGROUND: Patients diagnosed with pulmonary tuberculosis (TB) have poor sleep quality due to multiple factors. We aimed to assess the sleep status and related factors of TB patients in Shenzhen, China. METHODS: A questionnaire survey was conducted on 461 TB patients hospitalized at Shenzhen Third People's Hospital from March 2021 to January 2022, and sleep quality was assessed using the Pittsburgh sleep quality index (PSQI). RESULTS: A total of 459 valid questionnaires were collected, and 238 of the 459 TB patients had general or poor sleep quality (PSQI > 5). Patients' gender, marriage, nutritional screening score, family atmosphere, fear of discrimination, fear of interactions, and the impact of the disease on their work life had significant effects on sleep quality (P < 0.05); PSQI scores of TB patients were negatively correlated with lymphocyte counts (r = - 0.296, P < 0.01), T-lymphocyte counts (r = - 0.293, P < 0.01), helper T lymphocyte counts (r = - 0.283, P < 0.01), killer T lymphocyte counts (r = - 0.182, P < 0.05), and were positively correlated with depression scores (r = 0.424, P < 0.01). Multivariable logistic regression analysis showed that male (OR = 1.64,95% CI 1.11-2.42, P < 0.05), unmarried (OR = 1.57, 95% CI 1.02-2.42, P < 0.05), NRS score grade 3(OR = 5.35, 95% CI 2.08-15.73, P < 0.01), general family atmosphere (OR = 2.23, 95% CI 1.07-4.93, P < 0.05), and the disease affecting work (OR = 1.66, 95% CI 1.11-2.50, P < 0.05) were factors influencing poor sleep quality. CONCLUSION: Most TB patients had varying degrees of sleep disturbance, which may be affected by their gender, marriage, family atmosphere, nutritional status, the effect of the disease on work life, and, depression, as well as lower absolute T-lymphocyte subpopulation counts. Appropriate interventions should be implemented to improve their sleep quality, when treating or caring for such patients.

Effect of panel reactive antibodies on T cell immunity reinstatement following renal transplantation.

BACKGROUND: Chronic kidney disease is associated with immunological disorders, presented as phenotypic alterations of T lymphocytes. These changes are expected to be restored after a successful renal transplantation; however, additional parameters may contribute to this process. AIM: To evaluate the impact of positive panel reactive antibodies (PRAs) on the restoration of T cell phenotype, after renal transplantation. METHODS: CD4CD28null, CD8CD28null, natural killer cells (NKs), and regulatory T cells (Tregs) were estimated by flow cytometry at T0, T3, and T6 which were the time of transplantation, and 3- and 6-mo follow-up, respectively. Changes were esti mated regarding the presence or absence of PRAs. RESULTS: Patients were classified in two groups: PRA(-) (n = 43) and PRA(+) (n = 28) groups. Lymphocyte and their subtypes were similar between the two groups at T0, whereas their percentage was increased at T3 in PRA(-) compared to PRA(+) [23 (10.9-47.9) vs 16.4 (7.5-36.8 µ/L, respectively; P = 0.03]. Lymphocyte changes in PRA(-) patients included a significant increase in CD4 cells (P < 0.0001), CD8 cells (P < 0.0001), and Tregs (P < 0.0001), and a reduction of NKs (P < 0.0001). PRA(+) patients showed an increase in CD4 (P = 0.008) and CD8 (P = 0.0001), and a reduction in NKs (P = 0.07). CD4CD28null and CD8CD28null cells, although initially reduced in both groups, were stabilized thereafter. CONCLUSION: Our study described important differences in the immune response between PRA(+) and PRA(-) patients with changes in lymphocytes and lymphocyte subpopulations. PRA(+) patients seemed to have a worse immune profile after 6 mo follow-up, regardless of renal function.

A new indicator: The diagnostic value of CD8+T/B lymphocyte ratio in sepsis progression.

OBJECTIVE: To reveal the value of single lymphocyte subpopulation and their ratios in the progression of sepsis. METHODS: From January 2019 to March 2021, 39 sepsis patients, 16 septic shock patients, and 50 healthy volunteers were recruited in the Second Xiangya Hospital for this cross-sectional study. The absolute quantitation of CD4+T, CD8+T, B lymphocytes, and NK cells in peripheral blood were determined by flow cytometry. SPSS Software was used to analyze the results. RESULTS: On the whole, the numbers of lymphocytes in the sepsis group and in the septic shock group were lower than that in the healthy control group. Surprisingly, the percentage of CD8+T lymphocytes in the septic shock group was slightly higher than that in the sepsis group. The percentage of B lymphocytes in the sepsis group was higher than that in the healthy control group. The AUC of CD8+T/B was 0.724, with the sensitivity and specificity being 75.00% and 71.79%, respectively. CONCLUSION: The immune expression pattern of patients with sepsis was not a simple decrease in the number of lymphocytes. The change in the ratios of lymphocyte subpopulation might be more meaningful along the development and progression of sepsis. The ratio of CD8+T/B could be used to diagnose the progression of sepsis and reduce the misdiagnosis rate to a certain extent.

Changes of T lymphocyte subpopulations and their roles in predicting the risk of Parkinson's disease.

T lymphocytes are involved in the pathogenesis of Parkinson's disease (PD), while the heterogeneity of T-cell subpopulations remains elusive. In this study, we analyzed up to 22 subpopulations of T lymphocytes in 115 PD patients and 60 matched healthy controls (HC) using flow cytometry. We found that PD patients exhibited decreased naïve CD8+ T cells (CD3+ CD8+ CD45RA+ CD45RO-) and increased late-differentiated CD4+ T cells (CD3+ CD4+ CD28- CD27-), compared to HC, which were not affected by anti-parkinsonism medication administration. The proportion of naïve CD8+ T cells in PD patients was positively correlated with their severity of autonomic dysfunction and psychiatric complications, but negatively associated with the severity of rapid eye movement and sleep behavior disorder. The proportion of late-differentiated CD4+ T cells was negatively correlated with the onset age of the disease. We further developed individualized PD risk prediction models with high reliability and accuracy on the base of the T lymphocyte subpopulations. These data suggest that peripheral cellular immunity is disturbed in PD patients, and changes in CD8+ T cells and late-differentiated CD4+ T cells are representative and significant. Therefore, we recommend naïve CD8 + and late-differentiated CD4+ T cells as candidates for multicentric clinical study and pathomechanism study of PD.

Increased circulating innate lymphoid cell (ILC)1 and decreased circulating ILC3 are involved in the pathogenesis of Henoch-Schonlein purpura.

BACKGROUND: Innate lymphoid cell (ILC) dysfunction is involved in numerous immune diseases, but this has not been demonstrated in Henoch-Schonlein purpura (HSP). This study aimed to investigate whether ILC dysfunction or imbalance participate in the pathogenesis of HSP. METHODS: This was a prospective study in patients with HSP who were hospitalized at the Children's Hospital of Soochow University from June to December 2019. Age- and sex-matched controls were also enrolled. ILC subsets and lymphocyte subpopulations were determined by flow cytometry. The transmission immune turbidimetric method also facilitated the exploration of correlations between ILC subset frequency and lymphocyte subpopulation, as well as serum IgA in HSP patients. RESULTS: Fifty-one patients with HSP and 22 control patients were included. There were no differences in age and sex between the two groups. Compared with controls, patients with HSP had higher ILCs in relation to lymphocytes (P = 0.036), higher ILCs in relation to PBMCs (P = 0.026), higher ILC1s (P < 0.001), lower ILC3s (P < 0.05), and higher ILC1/ILC3 ratio (P < 0.001). Sixteen patients underwent routine therapy combined with methylprednisolone for 7-10 days; ILC1s were significantly decreased (P < 0.001) and ILC3s were increased (P = 0.033), and ILC1/ILC3 was significantly decreased (P < 0.001). Compared with the controls, the ratios of ILCs/lymphocytes and ILCs/PBMC were higher in patients in the arthritis and mixed groups (all P < 0.05). ILC1 were elevated in the purpura, arthritis, abdominal, and mixed groups (P = 0.027, P = 0.007, P < 0.001, and P < 0.001, respectively). ILC1/ILCs were positively correlated with CD3 + CD8 + T lymphocytes (r = 0.3701, P = 0.0075). The level of IgA did not correlate with ILCs. CONCLUSIONS: Higher circulating ILC1s and lower circulating ILC3s appear to be involved in the pathogenesis of HSP.

Effect of vitiligo treatment by compound Glycyrrhizin combined with fractional laser and Triamcinolone Acetonide injection on T Lymphocyte subpopulation.

OBJECTIVES: To discuss the effective mechanism of vitiligo treatment by compound glycyrrhizin combined with fractional laser and triamcinolone acetonide injection. METHODS: Forty-two patients with vitiligo vulgaris in the stable phase were classified into combined group (19 cases) and medicine group (23) admitted in dermatology department, Baoding First Central Hospital from January 2017 to July 2018. Both groups took 50mg compound glycyrrhizin orally three times per day, and applied halometasone cream externally once per day. Based on this treatment method, after the combined group adopted fractional laser, triamcinolone acetonide injection encapsulation was used immediately. After the treatment for six months, the curative effect was judged for both groups. Flow cytometry was used to test the changes of T lymphocyte subpopulation in peripheral blood before and after treatment. Meanwhile, immunohistochemical method was adopted to determine CD4 + and CD8 + T lymphocyte expression level. Besides, the normal control group was set up. RESULTS: The efficacy of combined group and medicine group were 73.68% and 56.52% respectively, P<0.05. The comparison of CD3 +, CD4 +, CD8 + and CD4 +/CD8 + T lymphocyte level in serum and skin damage before and after treatment had no statistical significance (P>0.05). Serum CD4 + T cells of vitiligo patients reduced, compared with the normal control group (P<0.05), and CD4 +/CD8 + declined (P<0.05). CD4 + and CD8 + T lymphocytes at the skin damage of patients increased, compared with normal control group (P<0.05). CONCLUSIONS: Compound glycyrrhizin combined with fractional laser and triamcinolone acetonide injection has good clinical effect in the treatment of vitiligo vulgaris in the stable phase, and its effective mechanism may have nothing to do with T lymphocyte subpopulation.

A low proportion of regulatory T cells before chemoradiotherapy predicts better overall survival in esophageal cancer.

BACKGROUND: Growing evidence indicates that survival is correlated with immune status in certain types of solid tumors. The immune system functions either to eliminate cancer cells or to keep cancer cells in check by maintaining an equilibrium; if there is a malfunction in the immune system, immune escape may happen which then allows cancer cells to grow into clinically apparent tumors. The progression of the tumor leads to a poor prognosis. Regulatory T cells (Tregs), a subset of CD4+ T cells plays a pivotal role in regulating the immune system. Tregs were identified as being CD4+ T cells expressing high levels of CD25. The purpose of this study was to investigate the influence of the proportion of pretreatment regulatory T cells (CD45+CD4+CD25HICD127LOW) on the prognosis of patients with non-operative chemoradiotherapy for esophageal cancer (EC). METHODS: A total of 297 patients with non-operative chemoradiotherapy EC treated at the Shanxi Province Cancer Hospital from January 2015 to January 2020 were included in the study. The correlation between regulatory T cell proportion and clinicopathological characteristics was analyzed by the chi-square test, while univariate survival analysis was carried out using the Kaplan-Meier method. Independent prognostic factors for overall survival (OS) were determined by univariate and multivariate Cox proportional regression models. RESULTS: Of the clinicopathological features analyzed, tumor location and the proportion of CD4+CD8+ double-positive cells were significantly associated (P<0.05) with the proportion of pretreatment regulatory T cells. An assessment of the prognoses showed that patients with a high proportion of regulatory T cells had a significantly worse OS than patients with a low proportion of regulatory T cells. The OS of patients with a low proportion CD4+CD8+ cells was lower than that of patients with a high proportion of CD4+CD8+ cells. A high proportion of CD4+CD8+ (>3.45%) cells and a low proportion of regulatory T cells (≤5.15%) would have better OS before chemoradiotherapy. CONCLUSIONS: Regulatory T cells play a role in predicting the prognosis of patients with EC before chemoradiotherapy.

EBV DNA increase in COVID-19 patients with impaired lymphocyte subpopulation count.

OBJECTIVES: The immunologic profile and opportunistic viral DNA increase were monitored in Italian patients with COVID-19 in order to identify markers of disease severity. METHODS: A total of 104 patients infected with SARS-CoV-2 were evaluated in the study. Of them, 42/104 (40.4%) were hospitalized in an intensive care unit (ICU) and 62/104(59.6%) in a sub-intensive care unit (SICU). Human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV), Parvovirus B19 and Human Herpesvirus 6 virus reactivations were determined by real-time PCR, and lymphocyte subpopulation counts were determined by flow cytometry. RESULTS: Among opportunistic viruses, only EBV was consistently detected. EBV DNA was observed in 40/42 (95.2%) of the ICU patients and in 51/61 (83.6%) of the SICU patients. Comparing the two groups of patients, the EBV DNA median level among ICU patients was significantly higher than that observed in SICU patients. In parallel, a significant reduction of CD8 T cell and NK count in ICU patients as compared with SICU patients was observed (p<0.05). In contrast, B cell count was significantly increased in ICU patients (p=0.0172). CONCLUSIONS: A correlation between reduced CD8+ T cells and NK counts, EBV DNA levels and COVID-19 severity was observed. Other opportunistic viral infections were not observed. The relationship between EBV load and COVID-19 severity should be further evaluated in longitudinal studies.

Changes in the Percentages of B- and T-Lymphocytes and Antibody Titres in Laying Hens Infested with Dermanyssus gallinae-A Preliminary Study.

(1) Background: Dermanyssus gallinae, a hematophagous ectoparasite, adversely affects the health status of laying hens, leading to reduced egg production and significant economic losses in commercial farms. The aim of this study was to determine the effect of D. gallinae on the development of post-vaccination immune responses in layer hens. (2) Methods: A total of 80 blood samples were collected at four time-points (B1-B4) from 10 Hy-Line Brown hens, randomly selected from a commercial layer farm. The flock was naturally infested with D. gallinae and treated twice with Biobeck PA 910 (AI silicon dioxide). The samples were collected before and after each treatment. The percentages of IgM+ B cells, CD3+/CD4+ T cells and CD3+/CD8a+ T cells were determined by flow cytometry; the titres of antibodies against avian encephalomyelitis, infectious bronchitis virus, Newcastle disease virus, Ornithobacterium rhinotracheale, reticuloendotheliosis virus and avian reovirus were determined by the immunoenzymatic method. (3) Results: The percentage of Th cells and post-vaccination anti-IBV and anti-NDV antibodies decreased significantly at the second infestation peak when the number of parasites was twice higher than at the first infestation peak. Non-significant negative correlations were found between the number of mites and the percentage of B cells (R = -0.845, p > 0.05) and between the number of mites and the percentage of Th cells (R = -0.522, p > 0.05), and a significant positive correlation was noted between the number of mites and the percentage of Tc cells (R = -0.982, p < 0.05). There were non-significant correlations between the number of mites and antibody titres. (4) Conclusion: The present findings suggested that D. gallinae might inhibit immune responses since the percentages of B cells and Th cells were negatively correlated with the number of mites. The percentage of Tc cells was positively correlated with the number of mites, which indicated that D. gallinae could stimulate cellular immune responses in infested laying hens. However, further research is needed to determine whether D. gallinae suppresses the production of vaccine-induced antibodies.

Lactobacillus fermentum Administration Modulates Cytokine Expression and Lymphocyte Subpopulation Levels in Broiler Chickens Challenged with Campylobacter coli.

This investigation was performed to assess the supplementation of probiotics on cytokine expression and lymphocyte subpopulation in Campylobacter coli challenged chickens. Thirty-six individuals were equally separated into four experimental treatments: C = untreated chickens, LB = probiotic control (Lactobacillus fermentum), Cc = Campylobacter-challenged control, LBCc = probiotic + Cc. All chicks were slaughtered and cecum samples were collected on day 4 postinfection. Gene expression analysis, using reverse transcription quantitative PCR (RT-qPCR), revealed significant differences in cytokine transcript expression between untreated and probiotic-treated chickens. In addition, flow cytometry was used to quantitate the levels of lymphocyte subpopulations. Principal component analysis showed that probiotic administration induced an overall downregulation of cytokine expression. C. coli exposure provoked a similar response to that of L. fermentum but to a lesser extent. Colonization of C. coli in the presence of the probiotic evoked a complex response with an upregulation of some type II cytokines, including interleukin IL-4 and IL-13, which could explain the increased presence of antibodies in both lamina propria and epithelium. Moreover, despite that the percentage of CD8 intraepithelial lymphocytes (IELs) was found to be higher, downregulation of proinflammatory cytokines IL-15, IL-16, and interferon γ was observed. This suggests that the detected CD8 are not effector cells but induced IELs, which release antimicrobial peptides, and are ready to be primed upon encountering antigen. These outcomes demonstrate that probiotic administration promotes a humoral response to a C. coli infection while dampening any potential inflammation mediated by effector T cells in 1-week-old chicks.

Manipulation of the immune system by non-small cell lung cancer and possible therapeutic interference.

Pulmonary carcinomas have developed mechanisms by which they escape the attack of immune cells. Immune checkpoint molecules programmed death 1 - programmed death ligand 1 (PD1-PDL1) and the cytotoxic T-lymphocyte antigen 4 system have gained attention. The expression of PDL1 by tumor cells causes immune tolerance, and further influences the microenvironment via orchestration by cytokines. Therapy with PDL1 antibodies could restore the cytotoxicity of T-lymphocytes towards tumor cells. Many patients will respond to this treatment. However, resistance mechanisms will counteract this therapy. New investigations have identified additional immune checkpoint inhibitors such as lymphocyte activation gene 3 and T cell immunoglobulin and mucin-domain containing-3. Tumor cells also induce tolerance by manipulating cells of the innate immune system. Macrophages are polarized to tumor-friendly M2, neutrophils into N2 types, and dendritic cells and myeloid suppressor cells are switched to assist tumor cells. Regulatory T cells enter the tumor microenvironment and signal tolerance to cytotoxic cells, inhibiting the influx of NK cells. Soluble mediators either released by tumor cells or cells of the tumor stroma induce immune tolerance, examples including tryptophan and indolamine dioxygenases, arginine and adenosine. Treatment options to counteract these molecules are currently being tested. The tumor stroma has been classified as immune-inflamed, immune-excluded, and immune-desert types. The latter might be switched to an inflamed type by induction of tertiary lymph follicles. Dendritic cells and macrophages normally phagocytose tumor antigens, but inhibitors of phagocytosis can block this. Interference with these molecules is another option for re-establishing the cytotoxic action of the immune system against tumor cells. In this review we will discuss these aspects with a special emphasis on non-small cell lung cancer.

Percentages of CD4+CD8+ Double-positive T Lymphocytes in the Peripheral Blood of Adults from a Blood Bank in Bogotá, Colombia

Objective: CD4+CD8+ double-positive T-cells (DPTs) have been classified as a separate T-cell subpopulation, with two main phenotypes: CD4high CD8low and CD4low CD8high. In recent years, the relevance of DPTs in the pathogenesis of infections, tumors, and autoimmune diseases has been recognized. Reference values among healthy individuals remain unknown. Therefore, the aim of this study is to provide a reference value for DPTs in peripheral blood from healthy donors in a blood bank in Bogotá, Colombia, and to determine the activation status using a surface marker. Materials and Methods: One hundred healthy donors were enrolled in the study. Peripheral blood cells were stained for CD3, CD4, CD8, and CD154 (CD40L), and cellular viability was assessed with 7-aminoactinomycin D and analyzed by flow cytometry. Results: The median value for DPTs was 2.6% (interquartile range=1.70%-3.67%). Women had higher percentages of DPTs than men (3.3% vs. 2.1%). The subpopulation of CD4low CD8high showed higher expression of CD154 than the other T-cell subpopulations. Conclusion: DPT reference values were obtained from blood bank donors. A sex difference was found, and the CD4low CD8high subpopulation had the highest activation marker expression.

Determinación y cuantificación de subpoblaciones de linfocitos T y células natural killer en sangre periférica de individuos sanos por citometría de flujo / Detection and quantification of T-cell subpopulations and NK cells in peripheral blood from healthy individuals

Resumen Introducción: La determinación de las diferentes subpoblaciones de los linfocitos T en las diversas patologías y el monitoreo postratamiento ayuda a que el médico tome decisiones terapéuticas teniendo como referencia la cinética de los linfocitos T localizados en sangre periférica. Métodos: Se realizó la estandarización de un perfil de moléculas de superficie para la caracterización de subpoblaciones de linfocitos T: naïve, activados y de memoria, así como las células natural killer o asesinas naturales (CD3− CD56+) en sangre periférica de individuos clínicamente sanos. Resultados: Se identificaron las subpoblaciones de linfocitos: naïve (CD3+, CD4+ o CD8+, CD45RA+, CD62L+, CCR7+), activados (CD3+, CD4+ o CD8+, CD45RA+ o CD45RO+, CD69+ y/o CRTAM+), efectores (CD3+, CD4+ o CD8+, CD45RA+, CD62L−, CCR7−), de memoria central (CD3+, CD4+ o CD8+, CD45RO+, CD62L+, CCR7+) y de memoria efectora (CD3+, CD4+ o CD8+, CD45RO+, CD62L−, CCR7−) en las poblaciones de linfocitos T CD4+ y CD8+. Se integraron los datos obtenidos con estadística descriptiva (valores mínimos, valores máximos, media, mediana). Conclusiones: Este panel será de gran utilidad para monitorear pacientes en quienes se requiera valorar el estado inmunológico desde el punto de vista celular. Particularmente, puede apoyar en el seguimiento de los pacientes en los que se requiera evaluar la reconstitución inmunológica (componente celular de estirpe T).

Abstract Background: The knowledge of the participation of different subpopulations of T lymphocytes in various pathologies helps to make therapeutic decisions, having as reference the presence of the different subpopulations of the T lymphocytes associated with the disease. Methods: A profile standardization of surface molecules for the characterization of subpopulations of T cells was conducted: naïve, activated and memory, as well as natural killer (CD3− CD56+) cells in peripheral blood of clinically healthy individuals. Results: Naïve (CD3+, CD4+ or CD8+, CD45RA+, CD62L+, CCR7+), activated (CD3+, CD4+ or CD8+, CD45RA+ or CD45RO+, CD69+ and/or CRTAM+), effectors (CD3+, CD4+ o CD8+, CD45RA+, CD62L−, CCR7−), central memory (CD3+, CD4+ o CD8+, CD45RO+, CD62L+, CCR7+), memory effectors (CD3+, CD4+ or CD8+, CD62RO+, CD62L−, CCR7−) subpopulations were analyzed by flow cytometry. Descriptive statistics parameters were calculated (minimum values, maximum values, mean values, median). Conclusions: This panel can be very useful for monitoring patients in whom the immunological status from a cellular perspective is needed. Particularly, it can support the follow-up of patients who require an immunological reconstitution (T-cell component) evaluation.