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PubMed1988

Class II (DR) antigen expression on CD8+ lymphocyte subsets in acquired immune deficiency syndrome (AIDS).

In a selected group of human immunodeficiency virus (HIV)-infected patients we confirm the expansion of a CD8+ T-lymphocyte subset, i.e., the CD8+/Leu7+ cells, which account for 30% of the lymphocytes, compared to 3% in the control donors. In addition, a CD8+ T-lymphocyte subset that coexpresses class II (DR) antigens, i.e., CD8+/DR+ cells, is also increased from 1.5% in controls to 27% in the HIV-infected patients. Using three-color immunofluorescence and flow cytometry we can demonstrate that the CD8+/Leu7+ and the CD8+/class II+ cells are not distinct but overlapping subsets. In the HIV-infected patients 42% of the CD8+/Leu7+ cells were strongly positive for class II and these CD8+/Leu7+/class II+ cells accounted for 13% of all lymphocytes. These findings indicate that the expanded CD8+/Leu7+ cells are activated and hence might be actively involved in immune defense in acquired immune deficiency syndrome (AIDS).

AIDS-Related Complex↗

PubMedDate not supplied

Effects of recombinant human erythropoietin on T lymphocyte subsets in hemodialysis patients.

Lymphocyte subpopulations were studied by immunofluorescence staining with monoclonal antibodies and laser flow cytometry in the blood of 23 hemodialysis patients before and after 3 months of treatment with recombinant human erythropoietin (rhEPO). Correction of anemia was accompanied by an increase in natural killer cells and a decrease in B lymphocytes. In the 11 patients (Group 1) with a baseline helper/suppressor (T4/T8) ratio greater than or equal to 2, the latter significantly decreased from 3.0 +/- 0.3 to 2.1 +/- 0.3 through both an increase in T8 cells and a decrease in T4 cells (p less than 0.005). Among the 12 patients with a pre-EPO T4/T8 ratio less than 2 (Group 2), no difference in T cell subsets was observed. The decrease in ferritin levels observed over the study period was not significant. In addition, the mean increase in hemoglobin levels during the first month of rhEPO therapy was greater in Group 2 than in Group 1 (1.1 +/- 0.3 vs. 0.6 +/- 0.3 g/dl, p less than 0.025). No change in any parameter was observed in eight control patients not receiving rhEPO. These results suggest that rhEPO can induce changes in lymphocyte subpopulations of hemodialysis patients through mechanism(s) yet to be clarified; conversely, the T4/T8 ratio might be a predictive factor for the erythropoietic response to rhEPO.

Anemia↗

PubMed1994

Lymphocyte subset abnormalities, autoantibodies and their relationship with HLA DR types in children with type 1 (insulin-dependent) diabetes and their first degree relatives.

Type 1 (insulin-dependent) diabetes mellitus is associated with abnormalities of circulating lymphocyte subsets and autoantibodies. To investigate the prevalence of these in non-diabetic siblings and non-diabetic patients of children with Type 1 diabetes, we analysed T-cell subsets of function and activation in 31 families with an index case of Type 1 diabetes and related these to autoantibodies and HLA DR type. Using two and three colour cytofluorimetry, we studied total and activated (HLA-DR+) CD3+, CD4+, CD8+, lymphocytes and on CD4+ lymphocytes the CD45RA/RO "naive" and "memory" cell phenotypes. Diabetic children (mean duration of disease 3.1 years) had a reduced total lymphocyte count (p < 0.05), their non-diabetic siblings a reduced CD4+ T-helper cell count (p < 0.05), and their parents a reduced percentage and number of CD3+ T cells (p < 0.01 and p < 0.05) compared with age-matched control subjects. Diabetic children, their siblings and parents all had significantly increased levels of activated CD4+ T-helper cells (p < 0.01, p < 0.05 and p < 0.01). In diabetic children and their siblings there was a significant over-expression of the CD45RO "memory" cell marker and significant under-expression of the CD45RA "naive" cell marker, whilst these were normal in the parents. Islet cell antibody positive diabetic children had significantly higher levels of CD45RO-expressing CD4+ lymphocytes than those who were islet cell antibody negative (p < 0.05). Amongst the siblings and parents, possession of HLA-DR4 was associated with lower percentages of CD4+ and higher percentages of CD8+ T cells. These findings extend current knowledge about the role of immunoregulatory CD45RA/RO cells in Type 1 diabetes. In addition, they demonstrate lymphocyte subset abnormalities in unaffected family members, some of which may be influenced by HLA DR alleles.

Adult↗

PubMed1990

Exercise and blood lymphocyte subset responses: intensity, duration, and subject fitness effects.

This study examined the effect of exercise intensity and duration on the percent blood lymphocytes in men of low [LF; maximal O2 uptake (VO2max) less than 50 ml.kg-1.min-1 and sedentary], moderate (MF; VO2max = 50-60 ml.kg-1.min-1 and recreationally active), and high (HF; VO2max greater than 60 ml.kg-1.min-1 and recent training history) fitness. Thirty healthy adult men (aged 20-31 yr) participated in four randomly ordered cycle ergometer rides: ride 1 (65% VO2max, 30 min), ride 2 (30% VO2max, 60 min), ride 3 (75% VO2max, 60 min), and ride 4 (65% VO2max, 120 min). Blood samples were drawn at various times before and after the exercise sessions. Lymphocyte subsets were determined by flow cytometry using monoclonal antibodies for total T (CD3+), T-helper (CD4+), and T-suppressor (CD8+) lymphocytes and for a subset of cells expressing a natural killer (NK) cell antigen (Leu7+). Plasma catecholamines were assayed to determine exercise stress. There were sharp reductions (P less than 0.01) in the percentage of pan-T and T-helper lymphocytes immediately after exercise across all fitness levels; the magnitude of this reduction was greatest after the highest intensity (ride 3) or longest duration (ride 4) work. In contrast, the absolute number of T and T-helper cells tended to increase after exercise and significantly so in the HF subjects (P less than 0.005). There was no significant effect of exercise or subject fitness category on the percentage of T-suppressor lymphocytes, although the absolute numbers of this subset increased significantly after exercise in LF subjects. Marked increases (P less than 0.01) in the percentage of NK cells occurred immediately after exercise at all intensities and durations tested; numerical increases in total NK cells were significant in all fitness groups after the highest intensity work (ride 3; P less than 0.005). Irrespective of whether the changes were expressed as percentage or total numbers, recovery to base line occurred at 30 min after exercise. The results suggest that the exercise effect on blood lymphocyte subset percentages in men is transient and occurs across all fitness levels. Concomitant changes in plasma catecholamine concentrations are only weakly associated with these lymphocyte subset percentage responses to exercise. Furthermore, this study shows that the exercise-induced changes in lymphocyte percentages do not consistently reflect changes in the absolute numbers of cells.

Adult↗

PubMed1994

Lymphocyte subset analysis by Boolean algebra: a phenotypic approach using a cocktail of 5 antibodies and 3 color immunofluorescence.

Commercial reagent kits for the evaluation of leukocyte subsets involve the staining of a panel of up to six tubes using combinations of pre-mixed fluorescein isothiocyanate (FITC) and R-phycoerythrin (PE) conjugated monoclonal antibodies. We describe a rapid method whereby total CD3+ T-cells, CD4+ T-cells (CD3+ CD4+), CD8+ T-cells (CD3+ CD8+), putative gamma delta-receptor-T-cells (CD3+ CD4- CD8-), and T-cells that are CD3+ CD4+ CD8+ as well as B-lymphocytes and NK-cells can be enumerated after staining in a single tube. Whole blood specimens are labelled with a mixture of antibodies: FITC-conjugated antibodies to CD4 and CD19, PE-conjugated antibodies to CD8 and CD16, and either peridinin chlorophyll protein (PerCP) or allophycocyanin (APC) labelling for antibodies to CD3. After recording 20,000 events the data were analysed on the Consort 32 computer system and LYSYS-II (Becton Dickinson, San Jose, CA) and all of the lymphocyte subset values were determined by Boolean algebra using a technique we refer to as Boolean gate analysis (BGA). Our study has shown that BGA is statistically equivalent to SimulSET lymphocyte subset analysis. Furthermore, the procedure reduces the number of tubes required to two with consequential saving in reagents, consumables, and time.

Antibodies, Monoclonal↗

PubMedDate not supplied

Different distributions of lung and blood lymphocyte subsets in pediatric AIDS or tuberculosis.

Pulmonary immunity has not been studied in children with acquired immunodeficiency syndrome (AIDS) or tuberculosis (TB), even though lungs of both children and adults infected with human immunodeficiency virus (HIV-1) or Mycobacterium tuberculosis are affected frequently and severely. In the present studies, the distributions of T (CD3+, CD4+, CD8+) and B (CD19+) lymphocytes in bronchoalveolar lavage fluid (BALF) and blood of children with AIDS (N = 28) and children with pulmonary TB (N = 18) were determined using direct immunofluorescence (flow microfluorimetry). The distributions of lymphocyte subsets in BALF differed dramatically from those in blood. In pediatric AIDS, reduction of CD4/CD8 ratio was much more pronounced in BALF than in peripheral blood (0.15 +/- 0.04 vs. 0.43 +/- 0.11). This difference was due to selective depletion of BALF CD4+ lymphocytes, rather than to a great influx of CD8+ cells into the lung. In childhood TB, the CD4/CD8 ratio in BALF also was significantly decreased, despite its elevation in blood (1.02 +/- 0.26 vs. 1.96 +/- 0.32). The results show that (1) examination of peripheral blood lymphocytes does not reflect the kind and extent of changes observed in the distribution of pulmonary lymphocyte subsets, and (2) the profound decrease of the CD4/CD8 ratios in BALF of children with AIDS or TB is due to decreased percentages and absolute numbers of BALF CD4+ lymphocytes. The data suggest that analysis of BALF provides a more accurate evaluation of the patient pulmonary immune status than monitoring peripheral blood.

Acquired Immunodeficiency Syndrome↗

PubMed1993

Changes in T and non-T lymphocyte subsets following seroconversion to HIV-1: stable CD3+ and declining CD3- populations suggest regulatory responses linked to loss of CD4 lymphocytes. The Multicenter AIDS Cohort Study.

We investigated changes in lymphocyte subsets (total, CD4+ and CD8+ T cells, as well as non-T cells) associated with human immunodeficiency virus type I (HIV-1) seroconversion in 321 homosexual or bisexual men in the Multicenter AIDS Cohort Study (MACS). These subjects had serial lymphocyte characterizations for up to 4 years before and 5 years after seroconversion. CD4+ lymphocyte levels declined rapidly in the first 18 months following seroconversion and less rapidly thereafter, while CD8 lymphocytes increased with similar kinetics. In contrast, total T (CD3+) lymphocytes declined only slightly in the first 18 months following seroconversion, and then remained stable. These results support the hypothesis of physiologic regulation of the total number of circulating T cells, such that lost CD4+ lymphocytes are replaced by newly generated CD4+ and CD8+ lymphocytes; over time, continued loss of CD4+ lymphocytes due to HIV-1 infection would result in net replacement of lost CD4+ lymphocytes with CD8+ lymphocytes. Non-T (CD3-) lymphocytes also declined after seroconversion, and this decline paralleled that of CD4+ lymphocytes. Thus, changes in both T and non-T lymphocytes after HIV-1 seroconversion may reflect the operation of homeostatic or regulatory mechanisms. Whether these mechanisms contribute to the development of immune deficiency requires further study.

CD3 Complex↗

PubMed1995

Allogenic BMT in children: differential lymphocyte subset reconstitution according to the occurrence of acute GVHD.

To acquire some biological markers associated with the occurrence of acute graft-versus-host disease (aGVHD) after allogeneic bone marrow transplant (BMT) in children, we have studied the lymphocyte subset reconstitution and the percentage of peripheral blood mononuclear cells bearing HLA-DR and HLA-DQ class II molecules. This study included 37 allogeneic BMT: either with (n = 17) or without (n = 20) aGVHD. Within 2 months after transplantation, we observed that patients with aGVHD had a unique mononuclear cell profile characterized by (i) a significant increase in the percentages of CD8bright+CD28- T cells (P = 0.05) and CD3+ T cells (P = 0.001), (ii) an important decrease in the percentage of CD56+ cells (P = 0.0001), and (iii) a decrease in the percentages of HLA-DQ+ and HLA-DR+ monocytes (P = 0.001) and HLA-DQ+ T lymphocytes (P = 0.0001), in comparison with patients without aGVHD. Moreover, statistical studies indicate that there was a positive correlation between CD8bright+CD28- and CD3+ T cells, whereas CD3+ T cells were negatively correlated to CD56+ cells. We did not find any statistical correlation between the percentages of HLA-DQ+ or HLA-DR+ cells and the percentages of these lymphocyte subsets. Therefore, in this study done in children, we suggest that patients with (i) less than 20% of DQ+ monocytes, (ii) less than 25% of CD56+ lymphocytes, and (iii) an enhanced percentage of CD8bright+CD28- T cells are strongly associated with aGVHD. Unfortunately, these biological markers of a GVHD may not precede the clinical manifestations of the disease.

Acute Disease↗

PubMed1994

[Changes in T-lymphocyte subsets of peripheral blood in unexplained habitual abortion].

UNLABELLED: T-lymphocyte subsets of peripheral blood were examined by flow cytometry (FCM) in 20 cases of unexplained habitual abortion (UHA) before and after active immunization treatment, with 10 normal pregnant (NP) women and 20 normal nonpregnant (NNp) women as control. THE RESULTS: there was no difference in the percentage of CD3 cells between the UHA cases and NNP. Before immunization the percentage of CD8 cells was significantly lower (P < 0.001) and the ratio of CD4/CD8 was increased (P < 0.05) in UHA as compared with the NNP. However, after immunization, the percentage of CD8 cells greatly increased (P < 0.05) and CD4/CD8 ratio decreased significantly (P < 0.05) nearly to that of NP. These findings suggest that: (1) the increase of CD4/CD8 ratio due to the decreasing CD8 cells is the underlying immunomechanism of UHA; (2) immunization may induce the proliferation of CD8 cells and the lowering of CD4/CD8 ratio. Therefore, CD8 cells might play an important role in maintaining the balance of fetal-maternal immunological relationship; (3) the ratio of CD4/CD8 may served as parameter for observing the clinical efficacy of immunization therapy.

Abortion, Habitual↗

PubMed1986

Autotransplantation of splenic fragments: lymphocyte subsets in blood, lymph nodes and splenic tissue.

Abnormal ratios of T helper-type to T suppressor-type lymphocytes in the blood of patients with replanted autologous splenic tissue led to the present study in rats. Lymphocyte subsets were studied in the blood, mesenteric lymph nodes and spleen after autotransplantation and compared to splenectomized and control rats. In the blood of transplanted rats the percentage of T and T helper-type lymphocytes was lower, in the spleen B lymphocytes higher and T lymphocytes and their subsets lower. Comparable changes were seen in the lymph nodes. The data of the mesenteric lymph nodes in autotransplanted rats did not differ from splenectomized animals. Even after 37 weeks the regenerated splenic tissue only reached 13% of the weight of control rats and the absolute lymphocyte number was only 2.5% of a normal spleen. Splenic autotransplantation results in a small hypocellular mass of splenic tissue with a different composition of lymphocyte subsets and does not correct the obvious effect of splenectomy on lymphocyte subpopulations in lymph nodes.

Animals↗

PubMed1987

Age-associated changes of T lymphocyte subsets.

The percentage and the absolute number of T3, T4, T6, T8, and T10 lymphocytes of the peripheral blood were determined in 100 children (3 days to 10 years), 29 adults (18 to 59 years), and 30 elderly individuals (60 to 98 years). The percentage of the various T lymphocyte subsets and T4/T8 lymphocyte ratio for the children and elderly did not differ statistically from adults, except for the proportion of T4 lymphocytes which was higher in the subgroup with 3 days of age. The absolute number of total lymphocytes, T3 and T8 lymphocytes was increased in children up to the age of 6 years while the number of T4 lymphocytes was increased up to the age of 2 years. Increased numbers of T6 lymphocytes were detected in children from 1 to 2 years of age while the number of T10 lymphocytes was increased in children up to the age of 2 years. Finally there was no difference between the results observed in elderly individuals and adults.

Adolescent↗

PubMed1994

Prostaglandin E2 levels and lymphocyte subsets in portal venous drainage of colorectal cancers.

Local immunosurveillance may play an important role in the growth and spread of tumors. To investigate the local immune response, we determined prostaglandin E2 (PGE2) levels (n = 23) and lymphocyte subsets (n = 10) in the portal vein draining the tumors of patients with colorectal cancer. PGE2 levels in the portal vein were significantly higher than those in the radial artery. Portal PGE2 levels in patients with advanced stage tumors (Duke's C and D) and lymph node metastases were significantly higher than those of patients with Duke's A and B tumors without lymph node metastases. Moreover, four of the nine patients with portal PGE2 levels greater than 100 pg/mL had liver metastases within 2 years of surgery. There were no differences in lymphocyte subsets between the portal vein and the radial artery. PGE2 levels in the portal vein draining colorectal tumor may thus be closely related to tumor progression and recurrence and may serve as a predictor of tumor recurrence in patients with colorectal cancer.

Adult↗

PubMed1983

Identification of lymphocyte subsets in the newborn using a variety of monoclonal antibodies.

Using a small sample of peripheral venous blood, the normal range for lymphocyte subpopulations (T-cell, B-cell, T-suppressor cell, and T-helper cell) was defined in non-infected preterm and term infants. Lymphocyte subsets were identified using a variety of monoclonal antisera, and analysis was performed using a fluorescent activated cell sorter. Such methods allow an objective assessment of absolute numbers of cells per unit volume of whole blood. There was no significant difference in absolute numbers of lymphocyte subsets between term and preterm appropriate for gestational age (AGA) infants. Infants who were small for gestational age (SGA) had a significant deficiency in absolute numbers of total T-cells, helper and inducer T-lymphocytes, and B-cells compared with both term and preterm AGA infants. All newborn infants (term and preterm; AGA and SGA) had a highly significant increase in absolute numbers of both helper and suppressor T-lymphocytes compared with normal adults.

Antibodies, Monoclonal↗

PubMed1993

Lymphocyte subsets in the mammary gland of sows.

The presence and localisation of lymphocyte subsets together with class II bearing cells in the mammary gland of sows, were studied at different periods of the reproductive cycle by immunohistochemistry and compared with blood. All cell types involved in the immune response were present in the mammary gland at the different stages of gestation and lactation and nearer the alveolar epithelium as gestation proceeded: T lymphocytes, including CD4+ and CD8+, B lymphocytes and class II bearing cells (epithelial cells and macrophages). The results indicated an early accumulation of T lymphocytes, specifically T helper cells, during pregnancy; the specific increase of IgA lymphocytes occurring after this phase could suggest a role for these T cells in the induction of IgA response. The local accumulation of immune cells sustains the view that the mammary gland is able to mount a true local immune response and the increase in CD8+ cells near the epithelium suggests a role in local immune defence.

Animals↗

PubMed1992

Lymphocyte subset and NK activity in cervical intra-epithelial neoplasia.

We have studied the systemic T-lymphocyte subset and, in particular, the natural-killer (NK) activity in 23 patients with cervical intra-epithelial neoplasia (CIN), and 11 controls (a history of normal cervical PAP smears and colposcopy patterns). The lymphocyte phenotypes were studied using a panel of monoclonal antibodies, and NK activity was evaluated as the percentage lysis of K 562 target cells. In patients with CIN we found a significant decrease in NK activity (p = 0.002) which was directly correlated with the grade of CIN, with no alterations in the absolute number of NK cells.

Adult↗

PubMed1991

Lymphocyte subset reference ranges in adult Caucasians.

We report here the distributions of lymphocyte populations bearing the following antigens: CD3 (T cells), CD19 (B cells), CD4 (T helper/inducer cells), CD8 (T suppressor/cytotoxic and some NK cells), and CD3-, CD16+, and/or CD56+ (NK cells). At four sites, analyses were performed on healthy, normal subjects between the ages of 18 and 70, using identical flow cytometry systems and techniques. Reference ranges (unadjusted for sex differences and age variation) are CD3 (61 to 85%), CD19 (7 to 23%), NK (6 to 29%), CD4 (28 to 58%), and CD8 (19 to 48%). The lymphocyte subpopulation distributions for all antigens were found to be similar at all sites. By combining data from all sites, it has been possible to estimate age variation and sex differences for each of these subpopulations. Age and sex associated differences are substantial for some lymphocyte subsets (CD3, CD4, NK cells), and proper accounting of these effects is essential in evaluating the individual patient, if further disease-related variation is to be accurately and consistently assessed. It appears possible to recommend reference ranges for lymphocyte population parameters applicable across national and laboratory boundaries. These ranges provide a basis for comparing results from different institutions and for combining such results on subjects and patients from several institutions, provided the methodology and equipment are identical at all sites.

Adolescent↗

PubMed1987

Lymphocyte subsets in urologic cancer patients.

Peripheral blood T lymphocyte subsets were quantitated by monoclonal antibodies in 91 patients with urologic malignancies as well as eight age-matched controls. In general, the presence of cancer and increasing stage were associated with a relative decrease in helper and an increase in suppressor T-cell activity as well as a decreased T4/T8 cell ratio. Therapy in the form of androgen depletion reversed these trends and restored helper and suppressor levels as well as the T4/T8 ratio to control values. In a small subset of sixteen patients surgery had only modest immediate (prostate) and long term (kidney) effects on these parameters.

Androgens↗

PubMed1995

Lymphocyte subsets and cell proliferation analysis in rabies-infected mice.

Alterations to lymphocyte subsets in the thymus and spleen of rabies-infected mice were investigated in a kinetic study covering the entire course of infection. Changes in the levels of thymic Thy1.2 cells were found to be proportional to total thymocyte depletion, while thymic CD4-/CD8- and B cell subsets were observed to increase in comparison to the total number of cells. The same was found to be true of CD4 and CD8 thymocytes, but only after the first clinical signs of disease had appeared. Meanwhile, drastic reductions in the number of CD4+/CD8+ thymocytes occurred during the course of infection. In the spleen, on the other hand, alterations to splenocyte subsets were not selective. Thymocytes expressing high levels of CD4 or CD8 markers were detected shortly before the death of the animals. Following the appearance of rabies symptomatology, lymphocyte proliferation decreased in comparison to the control. Thus, our results demonstrate that the thymus is the most injured lymphoid organ in cases of murine rabies infection.

Animals↗