Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Interleukin-7 Receptor alpha Subunit ; immunology ; Liver Cirrhosis, Biliary ; immunology ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo study changes to CD4(+)CD25(high+)CD127(low) regulatory T cells (Treg) in peripheral blood from children with bronchiolitis, and to explore its clinical significance.

METHODSThirty-one children with bronchiolitis and aged under two years were randomly enrolled as the bronchiolitis group, and 25 under two-year-olds with bronchopneumonia were randomly enrolled as the bronchopneumonia group. A further twenty-five children with non-infectious diseases such as hernia and renal calculus served as the control group. The level of CD4(+)CD25(high+)CD127(low) Treg in peripheral blood was measured by multi-color detection and multi-parameter flow cytometry.

RESULTSThe proportion of CD4(+)CD25(high+)CD127(low) Treg in peripheral blood in the bronchiolitis group (8.0%±2.1%) was significantly lower than in the bronchopneumonia (9.6%±2.6%; P<0.05) and control groups (11.3%±2.9%; P<0.05).

CONCLUSIONSCD4(+)CD25(high+)CD127(low) Treg level in peripheral blood may be an index of immunological function in infants. A decreased level of CD4(+)CD25(high+)CD127(low) Treg in peripheral blood suggests that Treg cells may be involved in the pathogenesis and development of bronchiolitis.

Bronchiolitis ; immunology ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Interleukin-2 Receptor alpha Subunit ; blood ; Interleukin-7 Receptor alpha Subunit ; blood ; Male ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo observe the proportion change of immune cells in the peripheral blood of patients with rectal cancer after neoadjuvant therapy and to explore the relationship between tumor regression and CD4⁺CD25(High)CD127(low) regularly T cells(Treg cells).

METHODSPatients with rectal cancer who underwent the neoadjuvant therapy before surgery at the Shanxi Cancer Hospital Colorectal Surgery Department from January to December 2013 were prospectively enrolled. These patients were divided into down-staging group and non-down-staging group according to the change of staging in accordance with TNM classification for rectal cancer after neoadjuvant therapy. Flow cytometry was used to analyze the proportions of Treg cells, CD4+T cells, CD8+T cells, NK cells, B cells, and CD4+/CD8+ ratio in the peripheral blood from these patients before and after neoadjuvant therapy.

RESULTSA total of 108 patients were enrolled, including 76 cases in the down staging group and 32 cases in the non-down-staging group. Differences of immune cells proportions between two groups before neoadjuvant therapy were not statistically significant(all P>0.05). In the down-staging group, the proportions of Treg cells, B cells and CD4+/CD8+ ratio were decreased while the proportion of NK cells did not change obviously after the neoadjuvant therapy. Interestingly, in the non-down-staging group, the proportions of B cells and CD4+/CD8+ ratio were decreased while the proportions of Treg cells and NK cells did not change obviously after the neoadjuvant therapy. In addition, after neoadjunvat therapy, the proportion of Treg cells in down-staging group was significantly lower than that in non-down-staging group [(4.4 ± 1.7)% vs. (6.2 ± 1.9)%, P=0.001].

CONCLUSIONFor patients in the down-staging group after neoadjuvant therapy, the proportion of Treg cells in peripheral blood decreases, suggesting that Treg cells may be a valuable biomarker for assessing tumor regression.

CD4-CD8 Ratio ; Flow Cytometry ; Humans ; Interleukin-2 Receptor alpha Subunit ; Interleukin-7 Receptor alpha Subunit ; Killer Cells, Natural ; Neoadjuvant Therapy ; Rectal Neoplasms ; T-Lymphocytes, Regulatory ; Treatment Outcome

OBJECTIVE: To investigate the IL-7R gene mutation and clinical features of adult patients with acute lymphoblastic leukemia (ALL). METHODS: One hundred sixty-four cases of newly treated adults with ALL from May 2016 to December 2018 were selected. Targeted and specific next-generation sequencing technology was used to detected a total of 16 types of Ph-like ALL mutations, which include IL-7R mutation, and the cilinical features, rate, types and sites of IL-7R were analyzed. RESULTS: IL-7R mutation was determined in 10 cases of 164 adult patients with ALL and the total mutation frequency was 13 times (6.1%). Out of 10 cases 5 cases were male (50%), 5 cases were female (50%). 6 cases of B-ALL ( 60% ) and 4 cases of T-ALL (40%). The mutation site of all cases was located at exon 6, among which 6 cases had replacement mutations, 3 cases had deletion mutations and 4 cases had insertion mutations. In addition, 1 triple and 1 double mutation of IL-7R were found. Besides, six mutation sites were newly identified, including: c.720_724del, c.723_726del, c.721_722insAGTG, c.727_728insTAACGGCCCCCTGCT, c.727_728insATGCAGGGAGCGAA and c.728_729insAAGTGTCA. CONCLUSION Six novel mutation sites and a poor manifestation of IL-7R have been explored in this research. Thus more samples are required to study the effects of IL-7R mutation on ALL treatment.
Adult ; Female ; Humans ; Interleukin-7 Receptor alpha Subunit ; genetics ; Male ; Mutation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Signal Transduction

OBJECTIVETo study the levels of CD4+CD25+CD127- and CD3+CD4-CD8- regulatory T (Treg) cells in peripheral blood of children with idiopathic thrombocytopenic purpura (ITP).

METHODSThe flow cytometry was used to detect the expression of CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells in peripheral blood of 33 children with ITP and 21 healthy children.

RESULTSThe expression levels of CD4+CD25+CD127-［(2.7±1.7)% vs (4.8±1.6)%; P<0.01］and CD3+CD4-CD8-［(5.2±3.1)% vs (8.1±3.5)%; P<0.01］Treg cells in children with ITP were significantly lower than in the controls.

CONCLUSIONSThe expression levels of CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells decrease in children with ITP, suggesting that CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells might play a role in the pathogenesis of ITP.

Adolescent ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Interleukin-2 Receptor alpha Subunit ; analysis ; Interleukin-7 Receptor alpha Subunit ; analysis ; Male ; Purpura, Thrombocytopenic, Idiopathic ; etiology ; immunology ; T-Lymphocytes, Regulatory ; immunology

The quantitative identification and enrichment of viable regulatory T cells (Treg) requires reliable surface markers that are selectively expressed on Treg. Foxp3 is the accepted marker of natural Treg, but it cannot be used to isolate cells for functional studies. CD127 is a new surface marker expressed in Treg cells. In this study, two populations of Treg, including CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+)T cells, and profiles of the Foxp3 expression in CD4(+)CD25(+)CD127(low/-) cells were compared to evaluate which population is better. The peripheral blood cells were collected and spleen suspension of BALB/C mice were prepared, and using triple staining CD4, CD25, CD127 and CD4, CD25, Foxp3. The profiles of Treg, including CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+) were detected by flow cytometry. The quadruple staining CD4, CD25, Foxp3 and CD127 were used to determine the CD127 expression in CD4(+)CD25(+)Foxp3(+) cells. The results showed that on T cell subset the median expression levels of CD4(+), CD4(+)CD25(+) were 39.02%, 5.35% in peripheral blood and 23.49%, 3.86% in spleen. On CD4(+) T cell subset, the median expression level of CD4(+)CD25(+)CD127(low/-) and CD4(+)CD25(+)Foxp3(+)T cells were 7.13%, 3.97% in peripheral blood and 12.8%, 8.23% in spleen. The ratio of CD4(+)CD25(+)CD127(low/-) T cells was higher than that of CD4(+)CD25(+)Foxp3(+) cells in both peripheral blood and spleen cells (P < 0.01). The CD4(+)CD25(+)CD127(low/-) cells highly expressed Foxp3, while the CD4(+)CD25(+)Foxp3(+)T cells lowly expressed CD127. It is concluded that compared with the CD4(+)CD25(+)Foxp3(+) populations, CD4(+)CD25(+)CD127(low/-) T cells better fit the definition of naturally occurring regulatory T cells in peripheral blood cells and spleen of BALB/C mice. CD127(low/-) is a characteristic marker on surface of CD4(+)CD25(+) Treg cells, and has been confirmed to be more specific marker for quantitatively sorting Treg cells.
Animals ; Biomarkers ; blood ; Female ; Interleukin-7 Receptor alpha Subunit ; analysis ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; T-Lymphocytes, Regulatory ; metabolism

OBJECTIVETo study the expression of CD4+ CD25int/high CD127low regulatory T cells in peripheral blood (PB) and its relation to the quantity of Hb, WBC and platelet (Plt) in children with aplastic anemia (AA).

METHODSExpression of CD4+ CD25int/high CD127low in PB was detected by flow cytometry in 22 children with AA before and after treatment and in 15 healthy controls. The relationships between CD4+CD25highCD127low and the quantity of Hb, WBC and Plt were evaluated.

RESULTSCompared to controls, the percentages of CD4+ CD25+/CD4+, CD4+CD25high/CD4+, CD4+ CD25+ CD127low/CD4+ and CD4+CD25highCD127low/CD4+ in PB of AA patients decreased markedly at the active phase (P﹤0.05). By the recovery phase, the percentages of CD4+CD25+/CD4+, CD4+CD25high/CD4+, CD4+ CD25+ CD127low/CD4+ and CD4+CD25highCD127low/CD4+ increased significantly to the levels similar to the controls. There were significant positive relationships between the expression of CD4+CD25highCD127low cells and the quantity of Hb, WBC and Plt (r=0.499, 0.526, 0.540 respectively; P﹤0.05).

CONCLUSIONSThe decrease of the percentage of CD4+CD25int/highCD127low regulatory T cells might be associated with the development of pediatric AA. The CD4+CD25int/highCD127low regulatory T cells can serve as a marker for the evaluation of disease severity as well as a target of further study on immune treatment of AA.

Adolescent ; Anemia, Aplastic ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Interleukin-7 Receptor alpha Subunit ; analysis ; Male ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo quantify the CD4+ CD25+ CD127(low) regulatory T cell (Treg), the expression levels of forkhead/winged helix transcription factor FOXP3 and Notch1 mRNA in aplastic anemia (AA) patients before and after treatment, and explore the significance of Treg in pathogenesis of AA.

METHODCD4+ CD25+ and CD4+ CD25+ CD127(low) T cells in peripheral blood were examined with FACS in 29 AA patients at active phase, 14 at recovery phase, 11 at unrecovery phase, and 15 normal controls. The levels of FOXP3 mRNA and Notch1 mRNA expression were detected with RT-PCR, and the correlations between Treg, FOXP3 mRNA and Notchl mRNA were analyzed.

RESULTSThe percentages of peripheral activated CD4+ CD25+ T cells in AA patients at active phase (4.3 +/- 0.7)% and unrecovery phase (4.2 +/- 0.6)% were significantly higher than those in normal controls (2.4 +/- 0.8)% (P < 0.05). The proportion of these cells in AA patients at recovery phase was reduced to (2.6 +/- 0.7)% (P < 0.05), being no difference from that in control group. The number of CD4+ CD25+ CD127(low) T cells in AA patients at active phase (2.4 +/- 1.2)% and unrecovery phase (2.5 +/- 1.1)% was decreased significantly compared with those in normal controls (7.1 +/- 2.7)% (P < 0.01) and in AA patients at recovery phase (5.3 +/- 1.0)% (P < 0.01), there was no difference between the latter two groups. In active phase AA patients, the levels of FOXP3 mRNA and Notchl mRNA (0.260 +/- 0.011 and 0.018 +/- 0.005, respectively) were lower than that in control group (1.307 +/- 0.011 and 0.308 +/- 0.028, respectively) (P < 0.01 and P < 0.01). After treatment, the levels significantly increased to 1.287 +/- 0.012 and 0.281 +/- 0.013 (P < 0.01 and P < 0.01), but there was no difference with that of normal controls (P > 0.05). CD4+ CD25+ CD2(low) T cells and FOXP3 were positively related with Notchl (P < 0.01) in AA patients.

CONCLUSIONThe decreased number and suppressive activity of CD4 CD25+ CD127(low) Treg cells in the peripheral blood of AA patients cause over-activation of autoreactive T cells and suppression of haematopoiesis. One of the mechanisms maybe the reduced expression of Notch1 in the target cells.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; immunology ; metabolism ; CD4 Antigens ; Case-Control Studies ; Female ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; Interleukin-7 Receptor alpha Subunit ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Receptor, Notch1 ; genetics ; metabolism ; T-Lymphocytes, Regulatory ; immunology ; Young Adult

BACKGROUNDThe frequencies of regulatory T cells (Tregs) increased over the HIV infection but its counts actually decreased. We proposed that the decrease of Treg counts may cause the reduction of inhibitory effect and thereby account for the over-activation of Tregs during HIV infection. However, it remains unknown whether Tregs are also over-activated and thereafter the activation induced death may lead to the decrease of Tregs.

METHODSTregs were defined as CD4(+)CD25(+)CD127(lo/-) T cells. Eighty-one HIV-1 infected patients were enrolled in our study, and twenty-two HIV-1 seronegative donors were recruited as the control. The levels of HLA-DR on Tregs were determined by FACSAria flow cytometer.

RESULTSCompared to HIV-1 seronegative donors, the levels of HLA-DR on CD4(+)CD25(+)CD127(lo/-) Tregs were significantly increased in HIV-1 infected patients, and its increase was positively associated with viral loads (r = 0.3163, P = 0.004) and negatively with CD4 T-cell counts (r = -0.4153, P < 0.0001). In addition, significant associations between HLA-DR expression on CD4(+)CD25(+)CD127(lo/-) Tregs and the percentages of HLA-DR, CD38, Ki67 expressing CD4(+) and CD8(+) T cells were also identified.

CONCLUSIONHLA-DR on Tregs is a good marker for viral replication and disease progression. The over-activation of Tregs might result in the decrease of Tregs.

ADP-ribosyl Cyclase 1 ; metabolism ; Adult ; Aged ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; Cells, Cultured ; Female ; Flow Cytometry ; HIV Infections ; immunology ; metabolism ; HLA-DR Antigens ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-7 Receptor alpha Subunit ; metabolism ; Lymphocyte Activation ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Young Adult

BACKGROUNDImmunosuppressive regulatory T cells (Tregs) participate in tumor immune evasion and the number and suppressive function of Tregs change with the aging process, but it is not clear whether such change leads to a higher incidence of tumors in the elderly. To this end, we designed experiments to explore the changes of Tregs and the functional gene Forkhead box P3 (FoxP3) in the aging process and its relationship with lung tumors in humans and mice.

METHODSThe percentage of CD4(+)CD25(+)CD127(low) Tregs and expression of FoxP3 mRNA were analyzed using flow cytometry (FCM) and real-time fluorescence-based quantitative polymerase chain reaction (FQ-PCR). Markers were analyzed in the peripheral blood (PB) of 65 elderly patients (age ≥ 65 years) with primary non-small cell lung cancer (NSCLC), 20 younger patients (aged < 55 years) with NSCLC, 30 elderly healthy individuals and 30 young healthy individuals. Furthermore, we set up the Lewis lung cancer model with C57BL/6 female mice. Thirty-six mice were divided into a young healthy group, a middle-aged healthy group, an elderly healthy group, a young tumor group, a middle-aged tumor group, and an elderly tumor group. The percentage of CD4(+)CD25(+)FoxP3(+) Tregs and the expression level of FoxP3 mRNA in splenocytes were determined in the six groups.

RESULTSThe percentage of peripheral CD4(+)CD25(+)CD127(low) Tregs and the expression of FoxP3 mRNA were significantly increased in elderly patients with NSCLC comparing with the other groups and in elderly healthy individuals compared with young healthy individuals. Further analysis showed that the percentage of CD4(+)CD25(+)CD127(low) Tregs and the expression of FoxP3 mRNA were closely associated with tumor node metastasis (TNM) staging in elderly patients with NSCLC. In the mouse model, the percentage of CD4(+)CD25(+)FoxP3(+) Tregs and the expression of FoxP3 mRNA in splenocytes of the tumor groups were significantly higher than in the healthy groups, with the highest expression in the elderly tumor group. In the healthy groups, the elderly healthy mice had the highest percentage of Tregs and expression of FoxP3 mRNA. The elderly mice had larger and heavier tumors than did the young and middle aged mice.

CONCLUSIONSThe up-regulation of Tregs and the FoxP3 gene with aging may play an essential role in oncogenesis and development of lung tumors in an elderly population.

Aging ; genetics ; metabolism ; Animals ; CD4 Antigens ; metabolism ; Female ; Flow Cytometry ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Interleukin-7 Receptor alpha Subunit ; metabolism ; Lung Neoplasms ; immunology ; metabolism ; Mice ; Mice, Inbred C57BL ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes, Regulatory ; immunology ; metabolism