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Comparison of low density lipoprotein uptake by different human lymphocyte subsets: a new method using double-fluorescence staining.

A method to determine low density lipoprotein (LDL) uptake of distinct lymphocyte subpopulations was developed using fluorescent LDL and subsequent staining of lymphocyte subsets with biotinylated monoclonal antibodies plus streptavidin-CyChrome. LDL uptake was detected on a single cell level and semiquantified by FACS analysis. This method allows comparison of defined lymphocyte subsets from different individuals and excludes the falsifying influence of individual differences in subset distribution, which may occur in studies on total peripheral blood lymphocytes (PBL). Investigation of total PBL and lymphocyte subsets of 20 healthy volunteers (8 male, 12 female) showed the following. i) Different lymphocyte subsets exhibited highly significant differences in LDL uptake, with NK cells (CD16) showing a higher uptake than T (CD3) and B cells (CD19); CD8-positive cells exhibited higher values than CD4-positive cells. ii) These differences are due to specific, LDL-receptor (LDL-R)-mediated LDL uptake. iii) Inter-individual differences in LDL uptake are reflected on all lymphocyte subsets.

Adult↗

Lymphocyte subsets recirculate from blood to lymph at different rates in conscious sheep.

Lymphocytes migrate and recirculate continuously between blood and lymph. The current study compared the migratory properties of lymphocyte subsets by labelling lymphocyte surface antigens and analysing simultaneous samples of mesenteric lymph, systemic arterial blood and mesenteric venous blood from conscious sheep. The lymphocyte output in lymph averaged 5.33 +/- 0.89 x 10(6) min-1 (lymph flow rate of 92 +/- 13 microliters min-1, lymphocyte count 61.9 +/- 10.7 x 10(9) l-1, n = 10) and the proportion of cells which carried the CD4 antigen (CD4+) was considerably higher in lymph (46 +/- 3%) than in arterial blood (23 +/- 2%; p < 0.01, n = 9). This increase was paralleled by a small but statistically significant decrease in the percentage of CD4+ cells in mesenteric venous blood (20 +/- 2%), as compared with systemic arterial blood (26 +/- 2%; p < 0.05, n = 6), which is consistent with the hypothesis that CD4+ cells migrate out of the blood vessels, and recirculate through the lymphatics, more readily than other lymphocyte subsets.

Animals↗

[RBC and T lymphocyte subsets in patients with bladder carcinoma: immunological study].

OBJECTIVE: To study immune function of erythrocytes and the correlation with the change of T lymphocyte subsets in the patients with bladder cancer. METHODS: The red blood cell C(3)b receptor rate (C(3)bRR), immune complex rate (ICR) and four kinds of tumor erythrocyte rosette tests were determined in 34 cases of bladder transitional epithelial carcinoma. The subsets of T lymphocyte were also determined by using flow cytometry (FCM). 30 non-carcinoma patients that well-matched in age and 20 healthy men were determined for comparison. RESULTS: The C(3)bRR, four kinds of tumor erythrocyte rosette rate, CD3, CD4 and CD4/CD8 ratio were lower in the bladder transitional epithelial carcinoma group than those of the control group. Meanwhile, ICR and CD8 were significantly elevated (P < 0.01). There was differentiation between G1 patients and G2 patients (P < 0.05). A positive correlation between C3bRR and the radio of CD4/CD8 was noted in the bladder transitional cell carcinoma group. CONCLUSIONS: Patients with bladder transitional cell carcinoma show lower erythrocyte function and disorder of T lymphocyte subsets. There is a close relationship between red cell immune function and T lymphocyte subsets. The cellular immune function (including the red cell immune function) in patients with poorly differentiated cells is lower.

Adult↗

Role of immunological lymphocyte subset typing as a screening method for lymphoid malignancies in daily routine practice.

BACKGROUND: The major diagnostic role of peripheral lymphocyte subset typing is to distinguish between malignant and reactive conditions. METHODS: The present study evaluates the screening efficacy of flow cytometric lymphocyte subset typing for the presence of a lymphoid malignancy. Four hundred samples were analyzed with a combination of anti-T-, B-, and natural killer (NK)-cell monoclonal antibodies. RESULTS: Two hundred and twenty (55%) samples showed a normal distribution of lymphocyte subsets, 73 (18%) samples exhibited unspecific alterations of lymphocyte subsets, 19 (5%) samples exhibited a reactive phenotype typical of Epstein-Barr virus/cytomegalovirus (EBV/CMV) infection, and 88 (22%) samples expressed a phenotype suggestive of lymphoma. The most predictive independent factor of a lymphoma-specific phenotype was the absolute lymphocyte count (P = 0.0001, odds ratio 73.225). Seventy-eight percent of samples containing >/=4 x 10(9)/l lymphocytes and 2% of samples with lymphocyte counts <4 x 10(9)/l exhibited a lymphoma-specific phenotype. The specificity of the referring clinical comment was the second best predictor of a lymphoma-specific typing outcome (P = 0.0001, odds ratio 19.589). The independent predictive values of lymphocyte morphology and of relative lymphocyte counts were of borderline significance. CONCLUSIONS: The use of flow cytometric lymphocyte subset typing as a diagnostic screening method for lymphoma should be restricted to cases of unexplained elevation of absolute lymphocyte counts with or without morphological atypias and to cases with definite clinical symptoms of lymphoma.

Cytomegalovirus Infections↗

Chemoattractant lymphokines specific for the helper/inducer T-lymphocyte subset.

The cellular content of T-lymphocyte-rich inflammatory sites is dependent in part on the in situ elaboration of chemoattractant factors. We have previously described three T-lymphocyte-specific chemoattractant lymphokines; a chemokinetic factor, lymphocyte chemoattractant factor (LCF, MW 56,000), and two distinct lymphocyte migration inhibitory factors (LyMIF75K, MW 75,000; and LyMIF35K, MW 35,000). These factors are produced by human T cells in response to antigen, concanavalin A, or histamine stimulation. In this communication, we report that LCF and LyMIF35K are produced by OKT8+ (suppressor/cytotoxic) and OKT4+ (helper/inducer) lymphocytes, respectively, and are selectively chemoattractant for the OKT4+ lymphocyte subset. LyMIF75K is produced by OKT4+ cells and inhibits both OKT4+ and OKT8+ lymphocyte migration. Production of LCF and LyMIF35K by infiltrating lymphocyte subsets may be one mechanism whereby unactivated helper/inducer T lymphocytes are selectively recruited to sites of inflammation.

Antibodies, Monoclonal↗

Bone marrow lymphocyte subsets in myelodysplastic syndromes.

AIM: To examine lymphocyte subsets in patients with myelodysplastic syndromes (MDS); and to correlate immunohistological variables with prognosis. METHODS: Bone marrow trephine biopsy specimens from 65 patients with MDS were immunophenotyped using a panel of antibodies. A minimum of 1000 cells from representative areas of marrow sections were counted at light microscopy. The association between immunohistological variables and prognosis was assessed. RESULTS: Compared with normal control marrows (n = 23) no major abnormalities of T cells (CD3), T cell subsets (CD4, CD8, CD25, TCR gamma/delta) or natural killer cells (CD56, CD57) were seen in the 65 patients. In high risk MDS (RAEB, RAEB-t) 19% of the cases showed increased numbers of B lymphocytes compared with none in the low risk group (RA, RARS) (p < 0.0090). Only percentages of B cells above 3% significantly correlated with poor survival (p = 0.0121 for CD19, p = 0.046 for CD22). CONCLUSIONS: The deviations in T lymphocyte counts seen in peripheral blood and in bone marrow aspirates could not be verified in bone marrow biopsy specimens.

Adolescent↗

Analysis of lymphocyte subsets in patients with aplastic anaemia.

Lymphocyte subsets have been measured in the blood of 28 patients with aplastic anaemia. The mean helper/inducer:suppressor/cytotoxic T lymphocyte ratio (1.24 +/- 0.74) was significantly decreased in the total population in comparison to mean ratios in a normal population (1.78 +/- 0.57) and in patients with other haematological diseases (1.82 +/- 0.92). A reversed ratio (less than or equal to 1) was present in a large proportion (53%) of aplastic patients, due both to an absolute deficiency of helper/inducer lymphocytes and an increase in suppressor/cytotoxic lymphocytes. Increased HLA-DR expression, evidence of lymphocyte activation, was present in seven of 12 patients evaluated and was confined to the suppressor/cytotoxic lymphocyte population. In the bone marrow, the percentage of lymphocytes was increased two-fold compared to normal bone marrow but the ratio of T cell subsets was not abnormal. Two of four patients evaluated after haematopoietic recovery following ATG treatment showed a return to normal of the T4/T8 ratio; in two others a low T4/T8 ratio persisted despite recovery. These results indicate that lymphocyte subset imbalance and lymphocyte activation are present in many patients with aplastic anaemia, either as a contributing factor or as a result of bone marrow failure.

Adolescent↗

Growth of Lucilia cuprina larvae following treatment of sheep divergently selected for fleece rot and fly strike with monoclonal antibodies to T lymphocyte subsets and interferon gamma.

Intensive lymphocytic infiltration of the underlying dermis occurs during cutaneous myiasis caused by larvae of the blow fly, Lucilia cuprina. To determine the effect of this infiltrate on larval growth, monoclonal antibodies (mAb) to CD4, CD8 or WC1 lymphocyte subset determinants were injected intravenously before and during experimental infection of sheep with larvae. The effect of intravenous injection of mAb to ovine interferon (IFN) gamma was also examined. The experiments were performed in 18-month-old maiden ewes with genetic resistance or susceptibility to the disease complex, bacterial dermatitis/cutaneous myiasis. mAbs induced profound depletion of CD8+ and WC1+ subpopulations from blood and skin at sites of larval growth. mAb to CD4+ gave only a moderate reduction in lymphocytes from blood or skin. mAb treatments did not modify larval growth or survival at 20 or 50 h after infection. Larval growth rates did not differ between resistant and susceptible genotypes. No evidence was found for a role of T lymphocyte subpopulations or the cytokine IFN, in modifying larval growth during the first 50 h of infection. It seems unlikely that T lymphocyte-dependent immunological effector mechanisms contribute to the lower prevalence of fly strike seen in the resistant genotype in the field.

Animals↗

[Relation of T-lymphocyte subsets and symptom complex groups of patients with chronic aplastic anemia].

The T-lymphocyte subsets of 38 patients with chronic aplastic anemia (CAA) were measured by monoclonal antibody (McAb) and ABC method and the relationship of T-lymphocyte subsets and symptom complex groups by traditional Chinese medicine theory were also analysed. It was shown that the T subsets of patients with CAA have remarkable changes: Ts (41.3 +/- 11.2%) increased more than the control (22.9 +/- 4.5%), Th/Ts decreased significantly more than the normal control (0.98 +/- 0.38 to 1.59 +/- 0.38). From the group of deficiency of both vital energy and blood to the group of deficiency of Kidney-yang to the group of deficiency of Kidney-Yin to the group of deficiency of both Kidney-Yang and Kidney-Yin, the Th lymphocyte gradually decreased, Ts lymphocyte successfully increased (from 33.8 +/- 7.9% to 57.7 +/- 7.5%), Th/Ts ratio declined (from 1.29 +/- 0.36 to 0.57 +/- 0.19). The Th/Ts ratio of the latter two groups decreased more than the former two groups (P less than 0.01). These results indicated that the T lymphocyte subsets and symptom complex group of patients with CAA were closely related and when deficient Yang affects Yin the immunologic function of body has a more obvious change. This revealed the connotation of Kidney-Yin and Kidney-Yang on the immuno-regulating cells (T lymphocyte subsets) level.

Adolescent↗

B and T lymphocyte subsets enter peripheral lymph nodes and Peyer's patches without preference in vivo: no correlation occurs between their localization in different types of high endothelial venules and the expression of CD44, VLA-4, LFA-1, ICAM-1, CD2 or L-selectin.

Many lymphocytes enter tissues such as peripheral lymph nodes, and Peyer's patches through high endothelial venules (HEV). It is known that HEV differ in the expression of adhesion molecules as lymphocyte subsets do. Through the interaction of these molecules B and T lymphocyte subsets are thought to be preferentially directed into lymphoid organs. However, it is unclear which role these mechanisms play in vivo, since there are no studies demonstrating that blood lymphocyte subsets preferentially interact with different types of HEV in vivo. Therefore, in the present study the frequency of B, T, CD4+ and CD8+ lymphocytes in the wall of the HEV of rat peripheral lymph nodes and Peyer's patches was analyzed by immunohistology. In addition, the expression of CD44, VLA-4, LFA-1, ICAM-1, CD2 and L-selectin on B and T lymphocyte subsets of the blood was determined by flow cytometry. Although B and T lymphocytes showed significantly different levels of expression for each adhesion molecule investigated, the relation of B and T lymphocytes within the HEV of peripheral lymph nodes and Peyer's patches was strikingly comparable (38.0 +/- 5.2% vs. 40.6 +/- 5.7% and 62.0 +/- 5.2% vs. 59.4 +/- 5.7%, respectively). The same was true for CD4+ and CD8+ cells. Thus, although HEV and the blood lymphocyte subsets differ markedly in their expression pattern of adhesion molecules, the existing levels are sufficient to mediate comparable entrance of B and T lymphocyte subsets into both types of HEV.

Animals↗

Lymphocyte subset numbers in cerebrospinal fluid: comparison of tick-borne encephalitis and neuroborreliosis.

OBJECTIVE: The aim of this study was to analyze lymphocyte subset numbers in cerebrospinal fluid (CSF) from patients with tick-borne encephalitis (TBE) and acute neuroborreliosis. METHODS: CSF lymphocyte subsets were enumerated in 42 TBE and nine neuroborreliosis patients using flow cytometry. RESULTS: The CSF numbers of CD4+, CD8+, HLA-DR+ and total-T lymphocytes, B lymphocytes, and NK cells were all greater in neuroborreliosis patients than in TBE patients. Neuroborreliosis patients showed positive correlation of CSF protein levels with the numbers of CD4+, HLA-DR+ and total-T lymphocytes. Also, the numbers of CSF B lymphocytes correlated positively with intrathecal Borrelia burgdorferi-specific IgG antibodies. Conversely, TBE patients demonstrated intrathecal protein levels that correlated positively with all investigated CSF lymphocyte subsets. CONCLUSION: These results suggest an intensive recruitment of lymphocyte subsets into the central nervous system (CNS) during acute neuroborreliosis, whereas TBE is characterized by a lower accumulation of lymphocyte subsets in the CSF.

Acute Disease↗

[Lymphocyte subsets in patients with recurrent upper airway infections].

UNLABELLED: The aim of our study was to evaluate the digressions of lymphocyte subsets in patients with recurrent upper airway infectious diseases. METHODS: We studied 35 patients (mean of age 11.1+/-2.1 years) with recurrent upper airway infections. The first group consisted of patients, who had acute upper airway infections: rhinitis, pharyngitis, laryngitis and tracheitis more than 6 times per last year, sinusitis or otitis more than 4 times per last year. The control group comprised of 9 healthy subjects. Subsets of lymphocytes (CD3+, CD4+, CD8+, CD4+/CD8+, CD16+/56+ and CD19+) were detected by FACS Calibur cytometer. RESULTS: We found a significantly lower count of CD4+ lymphocytes in the patients' group compared to the control group (37.5+/-1.2 vs 45.7+/-3.1% of total lymphocytes, p<0.01). We did not find any significant differences of other lymphocyte subsets between patients and control groups. CONCLUSION: We propose that patients with recurrent upper airway infections have alterations of the cellular immunity -- decreased amount of CD4+ lymphocytes.

Acute Disease↗

Recirculation of lymphocyte subsets (CD5+, CD4+, CD8+, SBU-T19+ and B cells) through gut and peripheral lymph nodes.

The surface phenotypes (CD5+, CD4+, CD8+, SBU-T19+, MHC class II+, T80+ and sIg+) of lymphocytes in blood, and in prescapular and ileocaecal efferent lymph of sheep, have been determined. The similarity in the distribution of lymphocyte subsets in both lymph compartments indicated that the non-random migration of lymphocyte populations to peripheral lymph nodes and gut could not be due to preferential migration of a particular lymphocyte subset such as CD4+ or CD8+ cells to one tissue. Marked differences in the percentage of lymphocyte subsets between blood and efferent lymph suggested that some lymphocyte subsets leave the blood with differing efficiencies and that differential extraction of lymphocyte subsets from blood by a lymph node may be due to subset-specific lymphocyte-endothelial interactions.

Animals↗

Effects of Nocardia rubra cell wall skeleton on lymphocyte subsets in former poison gas factory workers.

Peripheral blood lymphocyte subsets were determined in former workers of the Okunojima Poison Gas Factory (poison gas workers) having a high incidence of lung cancer, and the effect of administration of Nocardia rubra cell wall skeleton (N-CWS) was studied. In poison gas workers not receiving N-CWS, both the percentage and absolute number of Leu-2a+ cells were significantly increased as compared with normal controls, while both the percentage and absolute number of Leu-7+ cells as well as the Leu-3a/Leu-2a ratio were significantly decreased. In poison gas workers receiving N-CWS, the percentage of Leu-2a+ cells was significantly decreased when compared with poison gas workers not receiving N-CWS, while the percentage of Leu-1+ cells and both the percentage and absolute number of Leu-7+ cells as well as the Leu-3a/Leu-2a ratio were significantly increased. When N-CWS was administered to poison gas workers, the percentage of Leu-2a+ cells significantly decreased, with a minimum two weeks after administration, while the percentage of Leu-3a+ and the Leu-3a/Leu-2a ratio significantly increased and the percentages of Leu-1+ and Leu-7+ cells also increased. Furthermore, the absolute number of Leu-3a+ cells increased markedly with a peak two weeks after administration. The absolute numbers of both Leu-1+ cells and Leu-7+ cells as well as total lymphocytes also significantly increased, but that of Leu-2a+ cells showed hardly any change. It is considered that a single administration of N-CWS transiently corrects the abnormality of lymphocyte subsets, and that repeated administration of N-CWS once every three months may maintain the lymphocyte subsets of poison gas workers at normal levels.

Aged↗

Alterations of T-lymphocyte subsets, soluble IL-2 receptor, and IgE in peripheral blood of children with acute asthma attacks.

BACKGROUND: T-cell activation and alteration of cytokines are involved in the pathogenesis of atopic asthma. However, the profile of circulating T-lymphocyte subsets, related cytokines, and plasma IgE during acute asthma attacks is still unclear. OBJECTIVE: In an attempt to illustrate the dynamics of these parameters in asthma attacks, we investigated the changes of T-cell subsets, lymphocyte activation, soluble IL-2R, and IgE in peripheral blood in children during and after acute asthma attacks. METHODS: This study was carried out in a cohort of Chinese children (n = 59) with acute asthma attacks. Immunoassays were performed when the patients had acute attacks before treatment, and the patients were reexamined in the 4 weeks after the resolution of acute attacks with therapy. Paired t tests were used for the statistical analysis of these patients to compare the data obtained during and after the acute attacks. Twenty healthy, age-matched subjects were used as normal control subjects. Nine children with long-term stable asthma were used as control subjects with stable asthma. RESULTS: CD3+, CD4+, CD8+, and IL-2R+ (CD25+) cells; plasma soluble IL-2 receptor; and IgE were significantly higher in patients with acute attacks than in control subjects. (P <.05, P <.05, P <.001, P <.05, P <.0001, and P <.0001, respectively). Immunoelectron microscopy exhibited an increased expression of IL-2R on lymphocytes in acute attacks as compared to control subjects. The abnormalities returned to normal, with the exception of IgE, when clinical remission was achieved after treatment. Correlation analyses revealed a positive relationship between plasma IgE and soluble IL-2R in asthma attacks (r = 0.83, P =.0001). Plasma IgE and soluble IL-2R of those who were in remission positively correlated with their production in acute attacks (r = 0.58, P =.001 and r = 0.71, P =.0001, respectively). CONCLUSION: This study suggests that (1) the percentage of CD4+, CD8+, or IL-2R+ lymphocytes in peripheral blood was significantly elevated during acute attacks and returned to normal ranges after complete remission was achieved; (2) plasma soluble IL-2R is a sensitive marker for asthma activity; and (3) atopic asthmatic children seem to have a hereditary predisposition of having higher levels of soluble IL-2R in asthma attacks, coinherited with the trait of IgE.

Acute Disease↗

Platelet fatty acids and peripheral blood lymphocyte subsets in an institutionalized elderly population.

The associations between platelet fatty acids and peripheral blood lymphocyte subsets were studied in 78 institutionalized elderly individuals (27 men and 51 women), aged 67 to 100. Platelet fatty acids were assessed by gas chromatography, and peripheral blood lymphocyte subsets were quantitated by immunophenotyping using flow cytometry. It was found that women had a higher number of total T-cells (CD3), T-helper (CD3+4+) cells, and B-cells (CD19). However, no gender differences were observed in the percentages of lymphocyte subsets. In elderly men, after adjusting for age and fatty acid intake, the platelet concentration of omega-3 polyunsaturated fatty acids was positively related to the percentage of CD3 and CD3+4+ bearing lymphocytes (rs = 0.59, P < 0.05; and rs = 0.55, P < 0.05, respectively), and the concentration of total saturated fatty acids was also positively associated with the percentage of B (CD19) cells (rs = 0.50, P < 0.05). However, similar relationships were not observed in elderly women. No significant associations were found between trans fatty acids and any of the lymphocyte subsets in the study population. These findings suggest that fatty acids may be related to immune function. Any effects may be important in the host immune defence, especially in elderly individuals.

Aged↗

Changes of peripheral blood lymphocyte subsets before and after operation of patients with endometriosis.

This study was performed to elucidate whether endometriotic lesions can affect peripheral blood lymphocyte-subsets. Changes of lymphocyte-subsets of normal healthy women and patients with uterine myoma or endometriosis before and after operation were also examined by using two-color assay methods. The percentage and absolute number of CD57+CD16+ cells [moderately differentiated natural killer (NK) cells] of NK cell subsets in peripheral blood from patients with endometriosis were significantly lower than values from normal healthy women and patients with uterine myoma, while there was no difference in CD4+ and CD8+ lymphocyte-subsets between normal healthy women and patients with uterine myoma or endometriosis. In patients with endometriosis the percentage and absolute number of CD8+CD11+ cells (suppressor T-cells) was significantly increased after operation, while those in patients with urine myoma did not change. On the other hand, resection of endometriotic lesions resulted in a significant decrease of the percentage of CD57+CD16- cells (immature NK cells) and a significant increase of not only the percentage but also the absolute number of CD57+CD16+ cells (moderately differentiated NK cells), suggesting that existence of endometriotic lesions disturbs differentiation of the NK cells. Furthermore, suppressor inducer T-cells as shown by measuring CD4+2H4+ and CD4+4B4- cells were significantly increased after operation of endometriosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Lymphocyte subsets in the blood. The influence of splenectomy, splenic autotransplantation, ageing, and the site of blood sampling on the number of B, T, CD4+, and CD8+ lymphocytes in the rat.

Removal of the largest single lymphoid organ, the spleen, leads to an increase in severe infections. To prevent this, transplantation of splenic fragments can be performed, which may, however, cause an increase in CD8+ lymphocytes in the blood of these patients. This is controversial since in the clinical situation it is often difficult to account for the different age of the patients, the time point after the operation and many other factors known to influence the number of lymphocyte subsets. Using a well-defined animal model, B, T, CD4+, and CD8+ lymphocytes were determined preoperatively in adult rats. Then, either sham splenectomy, splenectomy, or splenic autotransplantation was performed and the animals were followed up for 15 months after the operation. The surgical procedure itself, the site of blood sampling and ageing all influenced the number of lymphocyte subsets profoundly. Furthermore, giving the data as relative or absolute numbers leads to different results. Splenectomy caused lymphocytosis, due to a significant increase in B and CD8+ lymphocytes, as did splenic autotransplantation, which indicates that the number of lymphocyte subsets in the blood should not be used to argue in favour of or against splenic autotransplantation. This study demonstrates that the number of lymphocyte subsets in the blood is influenced by many factors and therefore should be determined in a highly standardized fashion.

Aging↗