Peripheral Blood T-lymphocyte Subpopulations Research Articles

Relationship of thymic area with clinical-epidemiological variables and values of T-lymphocyte subpopulations in peripheral blood of children with recurrent infections

BackgroundRecurrent infections in childhood are the main cause of remission to the immunology service. T lymphocytes generated in the thymus are essential for fighting infection, making the thymus area an important predictor of the immune system’s competence. This study aimed to identify the possible relationship of the thymic area with clinical-epidemiological variables and values of subpopulations of T lymphocytes in the peripheral blood of children with recurrent infections.MethodsWe conducted applied research using a transversal analytical design at the National Medical Genetics Center (Havana, Cuba), from January to August 2022. The study covered 73 children of which we analyzed clinical-epidemiological variables and the size of the thymus through ultrasound. Furthermore, we determined the relative and absolute values of the subpopulations of T cells using flow cytometry.ResultsOf the children studied, 65.8% had thymic hypoplasia. The children who breastfed for less than 6 months showed four times the risk of developing moderate-severe thymus hypoplasia (OR = 3.90, 95% CI: 1.21–12.61). A direct relationship was found between the area of the thymus and the child’s size (r = 0.238, p = 0.043) and weight (r = 0.233, p = 0.047). The relative values of CD3+ T lymphocytes decreased in the cases of mild hypoplasia (p = 0.018) and moderate-severe hypoplasia (p = 0.049). The thymus area was associated with the absolute cell count of CD8+ effector memory T cells (rs = −0.263, p = 0.024) and of the central memory T cells (r = −0.283, p = 0.015).ConclusionsBreastfeeding for less than 6 months, as well as the weight and size of the child, are related to their thymus area. The subpopulation values of T lymphocytes detected suggest that patients with thymic hypoplasia develop a contraction of CD3+ T cells, which can make them more vulnerable to infectious processes. This finding was combined with an expansion of the memory compartments of the subpopulations of CD8+ T cells, suggesting a greater susceptibility to intracellular viral and bacterial infections in these cases.

Evaluation of peripheral blood T-lymphocyte subpopulations features in patients with hepatitis B virus-related acute-on-chronic liver failure based on single-cell sequencing technology

Objective: T lymphocyte exhaustion is an important component of immune dysfunction. Therefore, exploring peripheral blood-exhausted T lymphocyte features in patients with hepatitis B virus-related acute-on-chronic liver failure may provide potential therapeutic target molecules for ACLF immune dysfunction. Methods: Six cases with HBV-ACLF and three healthy controls were selected for T-cell heterogeneity detection using the single-cell RNA sequencing method. In addition, exhausted T lymphocyte subpopulations were screened to analyze their gene expression features, and their developmental trajectories quasi-timing. An independent sample t-test was used to compare the samples between the two groups. Results: Peripheral blood T lymphocytes in HBV-ACLF patients had different differentiation trajectories with different features distinct into eight subpopulations. Among them, the CD4(+)TIGIT(+) subsets (P = 0.007) and CD8(+)LAG3(+) (P = 0.010) subsets with highly exhausted genes were significantly higher than those in healthy controls. Quasi-time analysis showed that CD4(+)TIGIT(+) and CD8(+)LAG3(+) subsets appeared in the late stage of T lymphocyte differentiation, suggesting the transition of T lymphocyte from naïve-effector-exhausted during ACLF pathogenesis. Conclusion: There is heterogeneity in peripheral blood T lymphocyte differentiation in patients with HBV-ACLF, and the number of exhausted T cells featured by CD4(+)TIGIT(+)T cell and CD8(+)LAG3(+) T cell subsets increases significantly, suggesting that T lymphocyte immune exhaustion is involved in the immune dysfunction of HBV-ACLF, thereby identifying potential effective target molecules for improving ACLF patients' immune function.

Long-Term GAD-alum Treatment Effect on Different T-Cell Subpopulations in Healthy Children Positive for Multiple Beta Cell Autoantibodies.

Objective The objective of this study was to explore whether recombinant GAD65 conjugated hydroxide (GAD-alum) treatment affected peripheral blood T-cell subpopulations in healthy children with multiple beta cell autoantibodies. Method The Diabetes Prevention–Immune Tolerance 2 (DiAPREV-IT 2) clinical trial enrolled 26 children between 4 and 13 years of age, positive for glutamic acid decarboxylase autoantibody (GADA) and at least one other autoantibody (insulin, insulinoma antigen-2, or zinc transporter 8 autoantibody (IAA, IA-2A, or ZnT8A)) at baseline. The children were randomized to two doses of subcutaneously administered GAD-alum treatment or placebo, 30 days apart. Complete blood count (CBC) and immunophenotyping of T-cell subpopulations by flow cytometry were performed regularly during the 24 months of follow-up posttreatment. Cross-sectional analyses were performed comparing lymphocyte and T-cell subpopulations between GAD-alum and placebo-treated subjects. Results GAD-alum-treated children had lower levels of lymphocytes (109 cells/L) (p = 0.006), T-cells (103 cells/μL) (p = 0.008), T-helper cells (103 cells/μL) (p = 0.014), and cytotoxic T-cells (103 cells/μL) (p = 0.023) compared to the placebo-treated children 18 months from first GAD-alum injection. This difference remained 24 months after the first treatment for lymphocytes (p = 0.027), T-cells (p = 0.022), T-helper cells (p = 0.048), and cytotoxic T-cells (p = 0.018). Conclusion Our findings suggest that levels of total T-cells and T-cell subpopulations declined 18 and 24 months after GAD-alum treatment in healthy children with multiple beta-cell autoantibodies including GADA.

Immune profile of peripheral blood T-lymphocyte subpopulations in chronic hepatitis B

Background: The role of immune response to chronic hepatitis B virus (HBV) infection is complex; and the specific T-cell response to this infection can determine the duration and the extent of liver disease. Objective: This study aimed at assessing the profile of Tlymphocyte subpopulations in chronic hepatitis B (CHB) patients and its association with HBV replication. Methodology: The case group included 50 CHB patients with normal liver function tests (LFTs); and the control group included 50 age and sexmatched healthy individuals. The HBV markers, LFTs and serum viral load were measured in cases. Blood CD4 and CD8 T-lymphocyte subpopulations and the CD4/CD8 ratio were assessed in both groups by flow cytometry. Results: Our results showed significantly higher CD8 T-cells; significantly lower CD3 and CD4 T-cells; markedly reduced CD4/CD8 ratio in the cases as compared to the controls (P<0.001, for all). This T-cell impairment was also significantly linked to HBeAg positivity and elevated level of viraemia. The increased level of CD8 T-cells was significantly linked to both the HBeAg positivity (P<0.001) and the elevated level of viraemia (P=0.005), whereas the decreased levels of CD3, CD4 T-cells and CD4/CD8 ratio were significantly linked to both HBeAg positivity (P<0.001, in all) and the elevated level of viraemia (P<0.001, P=0.001& P=0.007, respectively). Conclusion: T-lymphocyte subpopulations imbalance could be expected by measuring the serum HBVeAg and the viraemia level in CHB patients exhibiting normal LFTs. These parameters are recommended to be measured regularly for the cellular immune function assessment.

Імунофенотип лімфоцитів крові у хворих на цукровий діабет 2-го типу з нормальною масою тіла та ожирінням

Актуальність. Визначення імунофенотипу лімфоцитів крові є одним із ключових показників функції імунітету у хворої людини. Однак дослідження імунофенотипу лімфоцитів у хворих на цукровий діабет 2-го типу (ЦД2) із найчастішим ускладненням при цьому захворюванні— надмірною масою тіла/ожирінням рідкісні та неоднозначні. Мета дослідження: визначення імунофенотипу лімфоцитів (CD3+ Т-, CD4+ Т-, CD8+ Т-, CD20+- і CD56+-клітин) крові у хворих з уперше виявленим ЦД2 із різним індексом маси тіла (ІМТ). Матеріали та методи. Обстежені 78 хворих з уперше виявленим ЦД2 і 40 нормоглікемічних осіб, які залежно від ІМТ були розділені на 4 підгрупи. Визначення імунофенотипу лімфоцитів крові було виконане проточно-цитометричним методом із використанням лазерного цитофлюориметра FACStar plus і панелі моноклональних антитіл до мембранних антигенів лімфоцитів. Результати. Для всієї групи хворих на ЦД2 характерне невелике, але вірогідне (p<0,05) підвищення абсолютної кількості CD4+ Т-клітин порівняно з групою нормоглікемічних людей. При поділі обстежуваних хворих залежно від ІМТ на 4 підгрупи: 1)≤ 25,5кг/м2; 2) 25,9–29,9 кг/м2; 3) 30,0–34,9 кг/м2; 4) > 35,0 кг/м2— виявлено, що у хворих першої підгрупи абсолютна кількість CD3+ Т-, CD4+ Т-, CD8+ Т-, CD20+- і CD56+-клітин була близькою до такої в нормоглікемічних осіб. У хворих другої підгрупи відзначалося вірогідне підвищення абсолютного числа CD4+ Т-клітин на 12,5% (p<0,05). У хворих третьої підгрупи спостерігали підвищення абсолютного числа CD4+ Т-клітин на 29,2% (p<0,001). У хворих четвертої підгрупи з морбідним ожирінням, особливо в жінок, було підвищення абсолютних чисел CD3+ Т- на 12,4% (p<0,01), CD4+ Т-— на 47,7% (p<0,001) і CD8+ Т-клітин— на 26,2% (p<0,001). Подібне підвищення абсолютного числа CD4+ Т-клітин залежно від ІМТ відзначалося також і в нормоглікемічних осіб, але було менш вираженим. Висновки. Для хворих з уперше виявленим ЦД2 характерне підвищення вмісту в периферичній крові субпопуляцій Т-лімфоцитів, особливо CD3+ Т- та CD4+ Т-клітин, найбільш виражене при супутньому ожирінні.

Clinical Significance of Composition Changes in T-cell Subpopulations After Chemotherapy in Patients With Gastric Cancer.

New therapeutic agents and prognostic biomarkers for gastric cancer are needed. We analyzed the composition of peripheral blood T-cell subpopulations in response to chemotherapy in patients with gastric cancer. Peripheral blood samples were collected from patients diagnosed with gastric cancer before and after chemotherapy (FOLFOX; oxaliplatin, 5-fluorouracil, and leucovorin). Peripheral blood mononuclear cells were isolated. Patients were divided into responder (n=5) and non-responder groups (n=2) based on their chemotherapy outcomes. Non-responders showed lower numbers of CD4+/total cells and CD8+/total cells after chemotherapy compared to the responder group, but the difference was not significant (p=0.905, p=0.095). Naïve T, central memory T, effector memory T and effector T-cell counts differed in both groups after chemotherapy. Changes in peripheral T-cell subpopulations after chemotherapy were confirmed in patients with gastric cancer, which may be a prognostic predictor and development of therapeutic agents.

Immunological effects of nivolumab immunotherapy in patients with oral cavity squamous cell carcinoma

BackgroundAlthough checkpoint blockades have become widely used, the immunological impact in cancer patients, especially those with oral cavity squamous cell carcinoma (OCSCC), has not been well studied.MethodsThe present study assessed the immunological impact of anti-PD-1 (nivolumab) treatment in 10 patients with OCSCC. This involved phenotypic analyses of peripheral blood T-cell subpopulations and their expression of immune mediators prior to and following nivolumab treatment. The focus was on immunological effects of treatment without regard to possible clinical responses.ResultsNivolumab caused a decline in the frequency of blood CD4+ cells but did not affect their expression of IFN-γ. However, nivolumab increased the proportion of CD4+ cells expressing the Treg-supporting factor Foxp3. Nivolumab treatment caused an increase in the proportion of CD8+ cells. While their expression of granzyme B increased, it did not attain significance. Analyses of CD8+ cell subpopulations showed nivolumab caused an increase in levels of unconventional CD8dimCD3+ T-cells. It also caused an increase in expression of granzyme B by these unconventional T-cells as well as by the conventional CD8hiCD3+ cells. The CD8hiCD3+ subpopulation also had a near-significant increase in IFN-γ expression. Treatment with nivolumab had no effect on the levels of the NK containing CD8dimCD3− subpopulation of cells or their expression of IFN-γ or granzyme B.ConclusionsThese results show nivolumab causes opposing effects on CD4+ and CD8+ cell populations, with CD4+ cell levels declining but increasing the proportion of Treg cells, and unconventional CD8+ T-cell levels increasing with increased expression of immune mediators by CD8+ T-cell subpopulations.

Disrupted balance of CD4+ T-cell subsets in bone marrow of patients with primary immune thrombocytopenia.

Disequilibrium of CD4+ T-cell subpopulations in peripheral blood (PB) of patients with primary immune thrombocytopenia (ITP) has been well established, whereas the profile of CD4+ T-cell subpopulations in bone marrow (BM) remains elusive. In the present study, the frequencies of T helper 22 (Th22), Th17, Th1, Th2, follicular T helper (Tfh) cells and regulatory T cells (Tregs) as well as their effector cytokines in BM and PB from active ITP patients and healthy controls (HCs) were determined. Results showed that the frequencies of Th22, Th17, Th1, and Tfh cells were significantly higher, but Treg number was remarkably lower in BM from ITP patients than from HCs. In the ITP group, it was notable that the numbers of BM Th22, Th17, Th1, Th2, and Tfh cells were significantly elevated compared with the matched PB counterparts, while Treg number in BM was considerably reduced compared with that in PB. In consistence with the BM Th subset pattern, plasma levels of interleukin (IL)-22, IL-17A, and interferon (INF)-γ in BM from ITP patients were significantly increased compared with that from HCs. Therefore, the balance of CD4+ T-cell subsets was disrupted in both BM and PB of ITP patients, suggesting that this might play important roles in the pathophysiological process of ITP.

Peripheral blood CD8+ T-cell profiles in patients with psoriatic arthritis: a cross-sectional case-control study.

While CD4+ T-cells are traditionally regarded as the main pathogenic T-cell subpopulation in psoriatic arthritis (PsA), the role of circulating CD8+ T-cells remains poorly characterized. We evaluated the differential representation of CD8+ T-cell subpopulations in peripheral blood (PB) of PsA patients. CD8+IL-17+, CD8+IFNγ+ and CD8+IL-17-IL-22+ T-cells were evaluated by flow-cytometry in 25 consecutive PsA patients, 7 rheumatoid arthritis (RA) patients, 16 patients with psoriasis, and 26 healthy controls (HC). We observed a significant expansion of circulating IFN-γ producing CD8+ T-cells in PsA when compared to psoriasis [21.2 (6.9-55.8)% vs. 3.8 (0.7-11.8)%, p < 0.0001] and HC samples [21.2 (6.9-55.8)% vs. 4.05 (0.44-19.8)%, p < 0.0001]. A frequency of circulating IFN-γ producing CD8+T-cells ≥ 9% distinguished PsA from psoriasis patients with a specificity of 84% and a sensitivity of 87.5% [AUC = 0.9 (0.80-0.99), p < 0.0001]. In addition, we found a significant expansion of circulating IL-17 producing CD8+ T cells in RA patients when compared to PsA, psoriasis and HC samples. By contrast, there were no significant between-group differences in the prevalence of circulating IL-22 producing CD8+ T-cells. In PsA patients there was a significant correlation between number of swollen joints and frequency of circulating IFN-γ producing CD8+ T-cells, and between extent and severity of psoriasis and frequency of circulating IL-17 producing CD8+ T-cells. Circulating IFNγ-producing CD8+ T-cells are raised in PsA when compared to psoriasis, suggesting a potential pathogenetic involvement of CD8+ T-cells and IFNγ production in chronic joint inflammation and damage. The significant enrichment of circulating IL-17 producing CD8+ T-cells in RA when compared to PsA warrants functional characterization and confirmation in larger studies. We found no significant enrichment of circulating IL-22 producing CD8+ T-cells in PsA, RA and psoriasis.

Increased Numbers of CD4+CD25+ and CD8+CD25+ T-Cells in Peripheral Blood of Patients with Rheumatoid Arthritis with Parvovirus B19 Infection.

To investigate T-cell subpopulations in peripheral blood of human parvovirus B19 DNA-positive (B19+) and -negative (B19-) patients with rheumatoid arthritis (RA) and healthy persons. Blood samples were collected from 115 patients with RA and 47 healthy volunteers; 27 patients with RA and nine controls were B19+ Cluster of differentiation (CD) 4, 8, 25 and 45RA were analyzed on blood cells. CD25 expression on CD4+CD45RA+, CD4+CD45RA-, CD8+CD45RA+, CD8+CD45RA- subsets were analyzed by flow cytometry. The percentage of CD25low and CD25hi cells was increased on CD4+CD45RA+, CD4+CD45RA- T-cells and the percentage of CD25+ cells was increased on CD8+CD45RA+, CD8+CD45RA- T-cells of B19+ patients with RA in comparison with B19- patients and controls. Raised levels of CD4 and CD8 regulatory T-cells in B19+ RA patients could cause down-regulation of antiviral clearance mechanisms and lead to activation of persistent human parvovirus B19 infection in patients with RA.

Effect of T-lymphocyte subpopulations in peripheral blood on disease-free survival of patients with different molecular subtypes of breast cancer

Effect of T-lymphocyte subpopulations in peripheral blood on disease-free survival of patients with different molecular subtypes of breast cancer

Severe impairment of regulatory T-cells and Th1-lymphocyte polarization in patients with Gaucher disease.

We investigated peripheral blood T-lymphocyte subpopulations and intracellular expression of IFN-γ, IL-4, IL-10, and IL-13, by whole blood flow cytometry, in 22 type I Gaucher disease (GD) patients. Results were compared with those of 19 sex- and age-matched controls. Patients with GD exhibited decreased frequencies and absolute numbers of CD3+/CD4+ helper T lymphocytes (40.8 ± 9.8% vs. 49.4 ± 5.7%, p = 0.002, and 0.77 ± 0.33 vs. 1.04 ± 0.28 × 10(9)/μL, p = 0.011), as well as increased frequencies of CD3+CD8+ suppressor T lymphocytes (23.8 ± 8.0% vs. 18.4 ± 3.8%, p = 0.010), resulting in a significantly decreased CD4/CD8 cell ratio (p < 0.001). Moreover, they had significantly increased percentages of IFNγ-producing both CD4+ and CD8+ T cells (p = 0.0003 and p = 0.023, respectively), implying a TH-1 polarization pattern. Finally, patients with GD had decreased percentages and absolute numbers of CD4+CD25(dim) T lymphocytes (p = 0.033 and p = 0.007, respectively), of CD4+CD25(high) T lymphocytes (p = 0.039 and p = 0.016, respectively), and of CD4+CD25(high)FOXP3+ regulatory T cells (p = 0.036 and p = 0.019, respectively). Our results demonstrate that patients with GD have a significant numerical impairment of T-helper lymphocytes and a constitutive TH1 direction pattern of activation of both CD4+ and CD8+ cells, associated with a significant decrease of T-regs. Ineffective T-cell control may explain the chronic inflammatory reaction and the increased incidence of lymphoid malignancies, which have been repeatedly reported among patients with GD.

Different pattern of T-cell subpopulations in peripheral blood of patients with rheumatoid arthritis at various stages of disease development

The comparison of changes in peripheral T-cell subpopulations at different stages of rheumatoid arthritis (RA) development may be important to understand the pathomechanism and to elucidate the course of RA. So far, there have been no comprehensive studies regarding the proportions of T cells in early and long‑lasting RA. The aim of this study was to assess the proportion of the main peripheral T-cell subpopulations in patients at various stages of RA development. We enrolled 75 patients who were divided into 4 subgroups depending on the diagnosis: undifferentiated arthritis (UA), which later developed into RA (UA‑RA) and other diseases (UA‑non‑RA); clinically confirmed untreated RA; long-term treated RA; and the control group. Flow cytometry was used to assess T-cell subpopulations. Patients with clinically confirmed untreated RA differed (P <0.05) in the proportion of CD4+ T-cell subpopulations expressing activation markers compared with controls (CD69, CD25, HLA‑DR, CD95) and UA patients (CD95). Untreated RA patients had the highest proportion of regulatory CD4+ T cells compared with control and other groups. The percentage of CD28- T cells was higher only in the group with clinically confirmed RA but not in those with early RA (at the UA stage). The peripheral T lymphocyte phenotype in very early RA is not similar to that observed in clinically‑confirmed RA. Patients with a confirmed diagnosis of RA can be easily differentiated based on the absolute numbers of the main T-cell subpopulations; however, the percentage of the main T-cell subpopulations do not discriminate those patients in the UA cohort who will develop RA.

SAT0123 Reduction Of T-Cell Clonal Expansion in Rheumatoid Arthritis Patients After 12 Months of Abatacept Treatment.

BackgroundThe T-cell compartment of patients with rheumatoid arthritis (RA) is characterized by profound changes, including an acceleration of immune-senescence, reduced diversity of the T-cell repertoire, and expansion of clonal populations....

Genome-wide association study for T lymphocyte subpopulations in swine

BackgroundLymphocytes act as a major component of the adaptive immune system, taking very crucial responsibility for immunity. Differences in proportions of T-cell subpopulations in peripheral blood among individuals under same conditions provide evidence of genetic control on these traits, but little is known about the genetic mechanism of them, especially in swine. Identification of the genetic control on these variants may help the genetic improvement of immune capacity through selection.ResultsTo identify genomic regions responsible for these immune traits in swine, a genome-wide association study was conducted. A total of 675 pigs of three breeds were involved in the study. At 21 days of age, all individuals were vaccinated with modified live classical swine fever vaccine. Blood samples were collected when the piglets were 20 and 35 days of age, respectively. Seven traits, including the proportions of CD4+, CD8+, CD4+CD8+, CD4+CD8−, CD4−CD8+, CD4−CD8− and the ratio of CD4+ to CD8+ T cells were measured at the two ages. All the samples were genotyped for 62,163 single nucleotide polymorphisms (SNP) using the Illumina porcineSNP60k BeadChip. 40833 SNPs were selected after quality control for association tests between SNPs and each immune trait considered based on a single-locus regression model. To tackle the issue of multiple testing in GWAS, 10,000 permutations were performed to determine the chromosome-wise and genome-wise significance levels of association tests. In total, 61 SNPs with chromosome-wise significance level and 3 SNPs with genome-wise significance level were identified. 27 significant SNPs were located within the immune-related QTL regions reported in previous studies. Furthermore, several significant SNPs fell into the regions harboring known immunity-related genes, 14 of them fell into the regions which harbor some known T cell-related genes.ConclusionsOur study demonstrated that genome-wide association studies would be a feasible way for revealing the potential genetics variants affecting T-cell subpopulations. Results herein lay a preliminary foundation for further identifying the causal mutations underlying swine immune capacity in follow-up studies.

Antitumor and immunomodulating effects of polysaccharides isolated from Solanum nigrum Linne

We have examined the effects of the crude polysaccharides isolated from Solanum nigrum Linne (SNL-P) in vitro and in vivo against U14 cervical cancer. SNL-P showed no antiproliferative effects in vitro at a dose up to 1 mg/ml. In vivo administration with SNL-P (90, 180, 360 mg/kg b.w., p.o.) decreased the number of ascites tumor cells and prolonged the survival time of U14 cervical-cancer-bearing mice. FACScan flow cytometer analysis showed that most of the ascites tumor cells were arrested in G2/M phase of cell cycle and the ratio of CD4+/CD8+ peripheral blood T-lymphocyte subpopulations were restored following treatment of SNL-P. Furthermore, the treatment with SNL-P also caused a significant increment in IFN-gamma (p < 0.01, 90, 180 and 360 mg/kg b.w.) and a remarkable decrease in Il-4 (p < 0.01, 90, 180 mg/kg b.w.; p < 0.05, 360 mg/kg b.w.) by the method of ELISA. These data showed that SNL-P possess potent antitumor activity and SNL-P might exert antitumor activity via activation of different immune responses in the host rather than by directly attacking cancer cells on the U14 cervical cancer bearing mice. Thus, SNL-P could be used as an immunomodulator and an anticancer agent.

Impaired T-Lymphocyte Subpopulations and Deranged Profile of Immunologic Activation in Patients with Gaucher Disease Type I

Patients with Gaucher disease exhibit substantial evidence of impairment of their immune system, namely, increased serum levels of proinflammatory cytokines and immunoglobulins, and increased incidence of B-cell malignancies, such as non-Hodgkin's lymphoma, MGUS and multiple myeloma. We investigated peripheral blood T-lymphocyte subpopulations with dual color flow cytometry, as well as the direction of T-lymphocyte activation, by using intracytoplasmic immunostaining for IL-2, IL-4, IL-10 and IFN-gamma, on resting CD4+ and CD8+ T-lymphocytes and following activation with PMA- 1 with the presence of Brefeldin-A. Evaluations were performed on 16 patients with type I Gaucher disease and on 17 healthy controls. Patients had significantly decreased absolute lymphocyte count (1621±684 vs 2148±566/mm3, p=0.013), CD3+ (1197±478 vs 1508±431/mm3, p=0.045) and CD4+ T-lymphocytes (658±245 vs 945±253/mm3, p=0.021), but not CD8+ T-lymphocytes (491±331 vs 486±189/mm3, p: n.s.), resulting in a significant reduction of the CD4/CD8 ratio (1.59±0.68 vs 2.16±0.83, p=0.041). The populations of naive CD4+CD45RA+ and of memory CD4+CD45RO+ T-lymphocytes were also significantly decreased (218±128 vs 432±179/mm3, p=0.0005 and 484±185 vs 631±231/mm3, p=0.056 respectively), however, CD8+CD45RA+ and CD8+CD45RO+ subpopulations did nor differ significantly, when compared to controls. CD3−CD56+, but not CD3+CD56+ lymphocytes were also decreased (131±82 vs 199±97/mm3, p=0.037). Patients had higher percentages of CD8+ (29.2±9.7 vs 23.5±6.8%, p=0.042), CD8+CD45RA+ (22.1±6.2 vs 18.3±5.0%, p=0.046) and CD8+CD45RO+ T-lymphocytes (13.2±6.2 vs 9.6±3.7%, p=0.027), as well as of activated CD8+HLA-DR+ (0.93±0.68 vs 0.48±0.21%, p=0.008) and CD4+HLA-DR+ T-lymphocytes (1.77±0.93 vs 1.09±0.48%, p=0.008). Moreover, although both, the absolute number and the percentage of CD20+ B-lymphocytes were similar, patients exhibited significantly increased absolute number and percentage of CD5+CD20+ B-lymphocytes (1.63±0.55 vs 0.64±0.37% p=0.00002 and 29±20 vs 13±8/mm3, p=0.011, respectively). Finally, patients with Gaucher disease had significantly increased resting TH2-polarized CD4+T-lymphocytes (CD4+IL-10+: 0.41±0.29 vs 0.24±0.11%, p=0.045) and TH1-polarized CD8+ T-lymphocytes (CD8+IFNγ+: 0.15±0.07 vs 0.08±0.04%, p=0.005, CD8+IL10+: 0.22±0.08 vs 0.32±0.014, p=0.052, and IFNγ+/IL4+ ratio among the CD8+ population 2.54±2.1 vs 1.08±0.91, p=0.018). Following mitogenic activation a very significant impairment of obtaining the TH1 phenotype was observed (CD4+IL2+ lymphocytes 33.7±17.1 vs 65.4±6.1%, p<0.00001). The above findings suggest that in patients with Gaucher disease there is a significant numerical impairment of T-helper lymphocytes and a shift towards TH-2 direction of lymphocyte activation. These findings may explain the rarity of autoimmune manifestations despite the chronic inflammatory reaction, as well as the increased incidence of lymphoid malignancies, which has been reported among patients suffering from this disease.

Hepatitis B virus DNA is more powerful than HBeAg in predicting peripheral T-lymphocyte subpopulations in chronic HBV-infected individuals with normal liver function tests

To investigate the peripheral T-lymphocyte subpopulation profile, and its correlations with hepatitis B virus (HBV) replication level in chronic HBV-infected (CHI) individuals with normal liver function tests (LFTs). Frequencies of T-lymphocyte subpopulations in peripheral blood were measured by flow cytometry in 216 CHI individuals. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time PCR. Information of age at HBV infection, and maternal HBV infection status was collected. ANOVA linear trend test and linear regression were used in statistical analysis. CHI individuals had significantly decreased relative frequencies of CD3(+), CD4(+) subpopulations and CD4(+)/CD8(+) ratio, and increased CD8(+) subset percentage compared with uninfected individuals (all P < 0.001). There was a significant linear relationship between the load of HBV DNA and the parameters of T-lymphocyte subpopulations (ANOVA linear trend test P < 0.01). The parameters were also significantly worse among individuals whose mothers were known to be HBV carriers, and those having gained infection before the age of 8 years. In multiple regressions, after adjustment for age at HBV infection and status of maternal HBV infection, log copies of HBV DNA maintained its highly significant predictive coefficient on T-lymphocyte subpopulations, whereas the effect of HBeAg was not significant. HBV DNA correlates with modification in the relative T-lymphocyte subpopulation frequencies. High viral load is more powerful than HBeAg in predicting the impaired balance of T-cell subsets.

Effect of viral load on T-lymphocyte failure in patients with chronic hepatitis B.

To investigate peripheral T-lymphocyte sub-population profile and its correlation with hepatitis B virus (HBV) replication in patients with chronic hepatitis B (CHB). Distribution of T-lymphocyte subpopulations in peripheral blood was measured by flow cytometry in 206 CHB patients. HBV markers were detected with ELISA. Serum HBV DNA load was assessed with quantitative real-time polymerase chain reaction (PCR). The relationship between HBV replication and variation in peripheral T-cell subsets was analyzed. CHB patients had significantly decreased CD3+ and CD4+ cells and CD4+/CD8+ ratio, and increased CD8+ cells compared with uninfected controls (55.44 +/- 12.39 vs 71.07 +/- 4.76, 30.92 +/- 7.48 vs 38.94 +/- 3.39, 1.01 +/- 0.49 vs 1.67 +/- 0.33, and 34.39 +/- 9.22 vs 24.02 +/- 4.35; P < 0.001, respectively). Univariate analysis showed a similar pattern of these parameters was significantly associated with high viral load, presence of serum hepatitis B e antigen (HBeAg) expression, liver disease severity, history of maternal HBV infection, and young age at HBV infection, all with P < 0.01. There was a significant linear relationship between viral load and these parameters of T-lymphocyte subpopulations (linear trend test P < 0.001). There was a negative correlation between the levels of CD3+ and CD4+ cells and CD4+/CD8+ ratio and serum level of viral load in CHB patients (r = -0.68, -0.65 and -0.75, all P < 0.0001), and a positive correlation between CD8+ cells and viral load (r = 0.70, P < 0.0001). There was a significant decreasing trend in CD3+ and CD4+ cells and CD4+/CD8+ ratio with increasing severity of hepatocyte damage and decreasing age at HBV infection (linear trend test P < 0.01). In multiple regression (after adjustment for age at HBV infection, maternal HBV infection status and hepatocyte damage severity) log copies of HBV DNA maintained a highly significant predictive coefficient on T-lymphocyte subpopulations, and was the strongest predictor of variation in CD3+, CD4+, CD8+ cells and CD4+/CD8+ ratio. However, the effect of HBeAg was not significant. T-lymphocyte failure was significantly associated with viral replication level. The substantial linear dose-response relationship and strong independent predictive effect of viral load on T-lymphocyte subpopulations suggests the possibility of a causal relationship between them, and indicates the importance of viral load in the pathogenesis of T cell hyporesponsiveness in these patients.

Acute Influence of FK506 on T-Lymphocyte Populations of Peripheral Blood and Spleen in Rats

Backgrounds Tacrolimus (FK506) is an immunosuppressive agent used in clinical transplantation. We studied the acute impact of FK506 on T-lymphocyte populations of peripheral blood and spleen in rats. Methods The animals injected with FK506 (2 mg/kg) subcutaneously were sacrificed at 12, 24, or 48 hours after administration. CD3 +, CD4 + and CD8 + T-cell populations in peripheral blood and spleen were analyzed by FACS. Results After treatment with FK506, the proportion of CD4 + cells significantly increased in the spleen at 12 hours, decreasing to control levels at 24 and 48 hours. There was a slight increase in the CD4 + subpopulation in the peripheral blood at 12 and 24 hours and a significant increase at 48 hours. The CD8 + T-cell subpopulations in peripheral blood and spleen were stable during FK506 administration. Conclusions These results indicated that FK506 increased CD4 + spleen lymphocyte activation in a short time, inducing a subsequent increase in CD4 + cells subsets in peripheral blood.