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Lack of direct immunosuppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on human peripheral blood lymphocyte subsets in vitro.

A wide variety of immunosuppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in experimental animals has been documented. In contrast, the impact of dioxin on the human immune system remains controversial, although adverse health effects have been reported in humans after occupational or accidental exposure to dioxin. Recently, Neubert et al. (1991) found that a dose-dependent decrease of peripheral blood lymphocyte (PBL) subpopulations in humans and non-human primates, including helper-inducer/memory cells (CD4+CD29+) and B cells (CD20+) occurred in pokeweed mitogen (PWM) stimulated cultures at concentrations as low as 10(-12)-10(-14) M TCDD. Therefore, the direct effects of dioxin on human PBL subpopulations have been studied, in order to determine their usefulness as sensitive biomarkers for human dioxin exposure. Lymphocyte cultures from healthy individuals were treated with 10(-7) M-10(-14) M TCDD in the absence and presence of stimulation with pokeweed mitogen (PWM) or anti-CD3 monoclonal antibody (moAb; OKT3) for 3 days. Cytochrome P450 (CYP1A1) enzyme induction, one of the best studied direct biological effects of TCDD on numerous cell types, was assayed in parallel by ethoxyresorufin-O-deethylase (EROD) activity. Percentages of the different lymphocytes subsets, including CD2 (T cells); CD4; CD45 RA (suppressor-inducer/virgin T cells); CD4 CD29; CD8; CD19 (B cells) as well as interleukin 2 (IL-2) receptor (CD25) and class II antigen (HLA-DR) expression, were analyzed by flow cytometry. DNA synthesis was determined by 3H-thymidine uptake after 3 days of culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Potent immunogenicity of the B subunits of Escherichia coli heat-labile enterotoxin: receptor binding is essential and induces differential modulation of lymphocyte subsets.

The importance of receptor binding in the potent immunogenicity of Escherichia coli heat-labile enterotoxin B subunit (EtxB) was tested by comparing its immunogical properties with those of a receptor binding mutant, EtxB(G33D). Subcutaneous immunization of EtxB(G33D) resulted in 160-fold reduction in antibody titer compared with wild-type EtxB, whereas its oral delivery failed to provoke any detectable secretory or serum anti-B subunit responses. Moreover, the two proteins induced strikingly different effects on lymphocyte cultures in vitro. EtxB, in comparison with EtxB(G33D), caused an increase in the proportion of B cells, many of which were activated (CD25+); the complete depletion of CD8+ T cells; an increase in the activation of CD4+ T cells; and an increase in interleukin 2 and a decrease in interferon gamma. These data indicate that EtxB exerts profound effects on immune cells, suggesting that its potent immunogenicity is dependent not only on efficient receptor-mediated uptake, but also on direct receptor-mediated immunomodulation of lymphocyte subsets.

Adjuvants, Immunologic↗

Changes in mitogen responsiveness lymphocyte subsets after traumatic injury: relation to development of sepsis.

Head injury and multiple trauma patients were evaluated for mitogen responsiveness and lymphocyte subset frequencies within the first few days after injury. The profile obtained was compared to the patient's clinical course to see if there was a relation between early immune abnormalities and the subsequent development of unanticipated sepsis. Lymphocytes from multiple trauma patients were generally hyporesponsive to in vitro stimulation with a suboptimal dose of the mitogen phytohemagglutinin (PHA). In contrast, the response of head injured patients was comparable to that of the control group. There was a significant decrease in the relative number of multiple trauma patient's T4 (29.3 vs 48.6%) and T11 (48.9 vs 74.7%) positive populations (P less than 0.01). There was no change in the percentage of T8-positive cells (19.0 vs 20.5%). Patients with head injuries also had a decrease in T4-positive cells (35.9%). The percentage of cells with B cell and natural killer (NK) markers remained normal. Thus trauma patients appeared to have an increase in null cells. Six patients whose PHA responses were among the lowest developed sepsis early after trauma. The changes in subset distributions although possibly contributing to a decreased responsiveness did not predict the ability to respond to PHA or the development of sepsis.

Adult↗

Relationships over 1 year between lymphocyte subsets and psychosocial variables among adults with infection by human immunodeficiency virus.

To examine relationships between immune and psychosocial variables among adults infected with human immunodeficiency virus type 1, 221 subjects without acquired immunodeficiency syndrome were assessed for degree of depression, anxiety, psychiatric symptoms, social support, stressful life events, hardiness, hopelessness, bereavement, and intrusive and avoidant thoughts about acquired immunodeficiency syndrome. At entry, none of 22 psychosocial variables significantly correlated with lymphocyte subsets. Among subjects seen 6 and 12 months later, severity of physical symptoms was associated with greater emotional distress, but the CD4 cell count was predicted by neither clinical ratings of psychopathology and global functioning nor by standardized self-report measures of constructs used in psychoimmune research. We conclude that among our sample, physical symptoms contributed to emotional distress, but emotional distress did not contribute to the CD4 cell count, a marker of disease progression.

Adolescent↗

Lymphocyte subsets in renal carcinoma--a sequential study using monoclonal antibodies.

Using monoclonal antibodies in conjunction with flow cytometry, circulating lymphocyte subsets with distinct functions in the regulation of the immune response were enumerated in 32 patients with proven renal carcinoma. Analyses were performed at presentation and sequentially during the clinical course of the patients. Untreated patients with advanced disease had a deficit of T cells with the "helper/inducer" phenotype (Leu-3a+) and this resulted in abnormal T "helper/suppressor" (Leu-3a+/Leu-2a+) ratios. Following nephrectomy, performed in 26 patients, there was a significant increase in the number of T cells with the "helper/inducer" phenotype and a significant increase in T "H/S" ratios. Subsequent follow-up at a minimum of 2 months after nephrectomy showed that the increase in T cells with the "helper/inducer" phenotype was maintained (with the exception of 6 patients with disease progression) and was then accompanied by a significant increase of the T cell subset with the "suppressor/cytotoxic" phenotype (Leu-2a+). Pre-operative renal arterial embolisation resulted in an early transient lymphopenia. The response to embolisation combined with nephrectomy was little different when compared with nephrectomy alone. These observations represent a novel view of the immunosuppressive effects of renal carcinoma and their relation to anaemia and disease progression are discussed.

Adult↗

Lymphocyte subsets in dialyser eluates: a new parameter of bioincompatibility?

INTRODUCTION: During haemodialysis (HD), several adverse reactions in peripheral blood can occur, which have been attributed to the bioincompatibility of the dialyser membrane. Utilizing a dialyser elution technique, we have demonstrated that polymorphonuclear cells (PMN) manifested non-membrane dependent signs of activation during HD with cellulose triacetate (CTA), cuprammonium (CU) and polysulphone (PS) membranes. In the present study, we employed this elution technique to investigate the influence of HD with these membranes on lymphocytes. METHODS: Eight patients were studied during HD with CTA, CU, and PS dialysers in a randomized crossover design. Dialyser elution was performed after 3 h of HD. Besides total leukocyte count and differentiation, lymphocyte subpopulations and activation status in peripheral blood and dialyser eluates were analysed by flow cytometry. RESULTS: Only with CU was a significant leukocyte decrease observed in peripheral blood at 30 min (P<0.001). Neither the total number of lymphocytes nor the proportion of T(CD3+) and B(CD19+) cells had markedly changed after HD with either membrane. Meanwhile, all membranes induced a relative decline in natural killer cells -NK(CD3-/CD16+/56+)- at the end of dialysis, although this was only significant for CTA (P=0.04). As for the T-lymphocyte subsets, the proportion of CD4+ cells had markedly increased after three hours of HD with all three dialysers, CTA and PS being significant (P<0.05). Dialyser eluates contained 33.8-82.2 x 10(6) cells, CTA yielding the highest cell counts. The majority (81-91%) of the eluted cells consisted of PMN dialyser eluates versus peripheral blood: P<0.05), whereas only few lymphocytes were found (4-13%, absolute 2.6 x 10(6)). Lymphocyte subpopulations in dialyser eluates were comparable to peripheral blood at t 180 in case of CTA and CU. In contrast PS eluates contained significantly fewer T-cells (37%), but more B-cells (22%) and NK-cells (30%) in comparison with peripheral blood at 180 min (peripheral blood: 79, 6 and 16% respectively; P<0.05). The expression of activation markers on T-cells (HLA-DR, CD25) in dialyser eluates was comparable with peripheral blood. Conclusions. The absolute number of lymphocytes in dialyser eluates of CTA, CU, and PS dialysers was low (mean 2.6 x 10(6)) in comparison with peripheral blood (mean 1.4 x 10(9)/l). Whereas non-selective adhesion occurred in CU and CTA dialysers, a selective adhesion pattern of lymphocyte subpopulations was observed in case of PS, suggesting a difference in bioincompatibility. Apparent T-cell activation was not noted, either in peripheral blood or in dialyser eluates. Because PMN in the dialyser eluates of three different membranes showed similar activation patterns in a previous study, we hypothesize that eluted lymphocyte, rather than PMN, represent a preferable parameter of bioincompatibility.

Adult↗

T-lymphocyte subsets and HLA-DR-expressing cells in rejected human kidney grafts.

This communication describes an immunohistochemical analysis of rejected human renal allografts. T-lymphocyte subsets were identified in frozen tissue sections, by Leu 1 (anti-'pan' T lymphocytes), Leu 2a (anti-'cytotoxic/suppressor' T cells), and Leu 3a (anti-'helper/inducer' T cells) monoclonal antibodies. In addition, HLA-DR-positive cells were identified by simultaneous labelling with heterologous anti-HLA-DR antibodies. T cells dominated the cellular infiltrates in acute cellular rejection. Leu-3a-positive cells were more numerous than Leu-2a-positive cells. The Leu-3a-positive cells usually appeared in clusters, whereas the Leu-2a-positive cells appeared scattered in the tissue. HLA-DR-positive non-T cells were found within clusters of T 'helper/inducer' cells. The cell pattern shares many features with the findings in delayed-type hypersensitivity reactions.

Antibodies, Monoclonal↗

Peripheral blood lymphocyte subsets and serum immunoglobulins in Sheehan's syndrome and in normal women during the menstrual cycle.

OBJECTIVE: There has been increasing evidence on the mechanisms underlying the interactions between the neuroendocrine and the immune systems, particularly in animal models with relatively few information in the human. In this study, we evaluate the cellular and humoral immunity in female patients with hypopituitarism and in normal women throughout the menstrual cycle in an attempt to determine the role of pituitary and gonadal hormones on the immune system. DESIGN: Serum immunoglobulins, peripheral blood lymphocyte subsets, and serum hormones were measured in eight patients with postpartum pituitary necrosis (Sheehan's syndrome) and in six normal women along different phases of the menstrual cycle, taking advantage of the lack of pituitary function and the cyclic variations in serum hormones, respectively. RESULTS: Patients with Sheehan's syndrome had higher T lymphocytes (CD2), including helper (CD4) and suppressor (CD8) cell subpopulations and B lymphocytes (CD19) when compared with normal menstruating women. An increase of serum IgA concentrations was also observed. Normal women showed little non-statistically different changes along the menstrual cycle in peripheral blood cell parameters and in serum immunoglobulin levels. CONCLUSIONS: a) Hypopituitarism in humans, in contrast with the animal model, may associate with immune up-regulation at both cellular and humoral levels; and b) hormonal changes along the normal menstrual cycle probably do not influence in great extent the immune system.

Adult↗

Characterization of banked umbilical cord blood hematopoietic progenitor cells and lymphocyte subsets and correlation with ethnicity, birth weight, sex, and type of delivery: a Cord Blood Transplantation (COBLT) Study report.

BACKGROUND: The Cord Blood Transplantation (COBLT) Study banking program was initiated in 1996. The study goals were to develop standard operating procedures for cord blood (CB) donor recruitment and banking and to build an ethnically diverse unrelated CB bank to support a transplantation protocol. STUDY DESIGN AND METHODS: The hematopoietic progenitor cell (HPC) and lymphocyte subset (LS) content of approximately 8000 CB units were characterized, and these results were correlated with donor ethnicity, birth weight, gestational age, sex, and type of delivery. RESULTS: There was a significant correlation of CD34+ cell count with colony-forming unit (CFU)-granulocyte-macrophage (r=0.68, p<0.001), CFU-granulocyte-erythroid-macrophage-megakaryocyte (r=0.52, p<0.001), burst-forming unit-erythroid (BFU-E; r=0.61, p<0.001), and total CFUs (r=0.67, p<0.001). Nucleated red blood cell count was significantly correlated with total CD34+ (r=0.56, p<0.001), total CFU (r=0.50, p<0.001), BFU-E (r=0.48, p<0.001), and counts of CD34+ subsets (p<0.001). Caucasian ethnicity was significantly correlated with higher CD3+/CD4+, CD19+, and CD16+/CD56+ LSs. Furthermore, CD34+/CD38- and CD34+/CD61+ CB units (HPC-C) were significantly lower in African American and Asian persons compared to Caucasian and Hispanic persons. Male sex was associated with significantly fewer CD3+/CD4+, CD19+, and CD16+/CD56+ but increased CD3+/CD8+ LSs (p<0.001). Finally, cesarean section was associated with significantly higher total CFU and CD16+/CD56+ but lower CD3+/CD4+, CD3+/CD8+, and CD19+ LSs. CONCLUSION: These results provide a standard and range for uniformly processed HPC-C progenitor cells and LSs. CB progenitor cells and/or LSs may in the future predict for rapidity of engraftment, incidence of graft-versus-host disease, speed and quality of immunore- constitution, graft-versus-tumor effects, and/or success of gene transfection after CB transplantation.

Antigens, CD34↗

Distribution of T-lymphocyte subsets in different portions of sarcoid granulomas: immunohistologic analysis with monoclonal antibodies.

The numbers and the distribution of T-lymphocyte subpopulations in lymph node granulomas from 11 patients with sarcoidosis were studied in cryostat sections by an immunoperoxidase technique. Greater numbers of helper T lymphocytes (Leu-3+) were found at the periphery than in the central portion of the same granuloma. Most of the suppressor T lymphocytes (OKT8+) were present at the periphery of the granulomas. In addition, the Leu-3a/OKT8 ratio varied from 0.7 to 1.8 in the outer compartment, while in the central portion of the granuloma much higher values (3 to 20) were found. These different distribution patterns of T-lymphocyte subsets provide evidence for two different compartments with different immune reactions in sarcoid granulomas.

Adult↗

Combined immunohistochemical staining for surface IgD and T-lymphocyte subsets with monoclonal antibodies in human tonsils.

The aim of the present paper is to detect two different antigens simultaneously in a single slide. In cryostat sections of human tonsils, B-lymphocytes of follicle mantle-bearing surface IgD were immunostained with the alkaline phosphatase method using monoclonal anti IgD. The subsequent staining for T-lymphocyte subsets (T-helper and T-suppressor lymphocytes) was performed again with the alkaline phosphatase method using one of the monoclonal antibodies OKT 4, OKT 8, Leu 3a, Leu 2a. The best results with the alkaline phosphatase method were achieved using naphthol AS phosphate and Fast Blue BB for the revelation of the first antigen and naphthol AS-BI phosphate and diazotized New Fuchsin for the second.

Antibodies, Monoclonal↗

T-lymphocyte subsets in nude mice with Giardia muris infection.

The aim of this study was to compare the relative percentages of T-lymphocyte subsets in athymic (nude) mice and immunocompetent BALB/c mice, with and without Giardia muris infection. Suspension of mononuclear leukocytes from blood, spleen, and Peyer's patches of uninfected mice were incubated with fluorescent monoclonal antibodies (MAbs) directed against mouse T-lymphocytes (anti-Thy-1.2 MAb), T helper/inducer lymphocytes (anti-L3T4 MAb), or T suppressor/cytotoxic lymphocytes (anti-Ly-2 MAb), and examined by fluorescence microscopy to quantify each of these cell populations. While the percentages of Thy-1.2+, L3T4+, and Ly-2+ lymphocytes are all reduced in nude mice, L3T4+ lymphocytes were found to be especially depleted, resulting in a reversed L3T4+/Ly-2+ ratio (less than 1) in all of the nude mouse tissues examined. BALB/c and nude mouse Peyer's patch T-cell subsets were quantified at various times during enteric Giardia muris infection. Unlike immunocompetent BALB/c mice, nude mice have an impaired ability to clear this infection. It was found that clearance of G. muris infection is associated with a Peyer's patch L3T4+/Ly-2+ ratio of greater than or equal to 2, and that this infection does not alter the percentages of Peyer's patch Thy-1.2+, L3T4+, or Ly-2+ lymphocytes in BALB/c mice or nude mice. The data obtained in the present work and in other studies suggest that the impaired capacity of nude mice to clear G. muris infection results from deficiency of L3T4+ lymphocytes.

Animals↗

[Effects of different anesthesia methods on the T lymphocyte subsets and hemodynamics of patients with abdominal tumor in peri-operational period].

OBJECTIVE: To explore the effects of different anesthesia Methods on the cellular immune function and mean arterial pressure, heart rate in patients with abdominal tumor in the peri-operational period. METHODS: Patients were anesthetized by general anesthesia (GA), combined acupuncture-general anesthesia (AGA), combined epidural-general anesthesia (EGA), and combined acupuncture-epidural-general anesthesia (AEA), respectively. Changes of T lymphocyte subsets and hemodynamics were observed at different time points before and after treatment. RESULTS: Patients' cellular immune function after GA was lowered, which could be alleviated by combination of GA with acupuncture anesthesia (AA) or with epidural anesthesia. Combined acupuncture-drug anesthesia showed a stabilizing effect on hemodynamics in peri-operational period to some extent. CONCLUSION: EGA and AGA are good anesthesia choice in clinical practice, which would be favorable for early recovery of immune function in patients with abdominal tumor after operation.

Acupuncture Analgesia↗

T-lymphocyte subsets and interleukin-2 production in zinc-deficient rats.

1. It has been suggested that zinc-deficiency impairs cellular (T-lymphocyte-mediated) immune responses via a selective effect on helper T-lymphocytes. We have addressed this question in the rat by employing recently developed reagents in the form of monoclonal antibodies which specifically identify rat T-lymphocyte subsets (identifying total T-cells, helper T-cells and suppressor T-cells) and also by quantifying helper T-cell function by measurement of the helper T-cell-derived molecule interleukin-2 (IL-2). 2. Zn-deficiency induced T-cell atrophy (assessed morphologically and phenotypically with anti-rat T-cell monoclonal antibodies) in both peripheral blood and spleen. The use of these specific monoclonal antibodies failed to demonstrate a selective effect of Zn deficiency on the helper T-cell fraction of the total T-lymphocyte population. 3. In contrast, the results of functional assays of the T-lymphocyte response were dependent on the conditions of culture but suggested that the generation of IL-2 and its corresponding receptor were determined by the intracellular Zn status. Thus, in vivo, helper T-lymphocyte numbers are non-specifically reduced since other T-cell subsets are also reduced in response to appropriate stimulation. The functional consequences of this are dependent on the intracellular concentration of Zn but appear to influence both IL-2 production and its receptors on activated T-cells.

Animals↗

[Pathogenetic and clinical significance of peripheral activated T-lymphocyte subsets in Basedow's disease].

The peripheral activated T-cell subsets which express concurrently DR and CD antigens were determined in 22 patients with Graves' disease using the fluorescence-rosette double labelling technique. In the active phase of the disease a decrease in the percentage of CD8+ cells, an increase in the CD4+/CD8+ ratio and an elevation of various DR+CD+T-lymphocytes was found. A concurrent marked increase in DR+CD4+ cells accounts for an abnormality in the equilibrium of activated T-lymphocyte subsets and an increase in DR+CD4+/DR+CD8+ ratios. In comparison with the active phase the DR+CD3+ and DR+CD4+ cells decreased in remission, but DR+CD8+ cells remained unchanged. The decrease in the ratio of CD4+/CD8+ cells was less marked than that of the DR+CD4+/DR+CD8+ cells, which fell into the normal range. Our results suggest an excessive activation of T-helper cells as well as a deficiency in T-suppressor cells which may play an important role in the pathogenesis of Graves' disease.

Adult↗

Persistent long-term changes in lymphocyte subsets induced by polyclonal antibodies.

BACKGROUND: Clinicians are well aware of the short-term effects of immunosuppression by mono- or polyclonal antibodies. Little is known about long-term changes induced by these therapies. METHODS: Forty-three renal allograft recipients were selected according to their initial postoperative immunosuppression: (1) BI group=basic immunosuppression with steroids and cyclosporine, n=16; (2) ATG group=basic immunosuppression plus polyclonal antibody antithymocyte globulin (ATG), n=11; and (3) OKT3 group=basic immunosuppression plus monoclonal antibody OKT3, n=16 patients. At intervals of 6 months, the following parameters were measured prospectively: lymphocyte surface antigens (HLA-DR, CD3, CD4, CD8, CD16, CD19, CD56, and CD57); serum and urine neopterin; serum amyloid A; and indirect and direct tests for herpes viruses. RESULTS: The mean period of observation was 58.4 months. The most significant differences between the groups occurred for CD4+ and CD8+ T cells. The ratios of CD4+ to CD8+ cells (n=278 measurements) were significantly and persistently lower in the ATG group (P<0.001, Brown-Mood test). Five years after transplantation, the ATG group had a CD4+ to CD8+ cell ratio of x=0.6 versus x=1.7 in the OKT3 group and x=2.0 in the BI group. This inversion was due to a persistent depletion of the CD4+ cells and an increased regeneration of the CD8+ cells, in particular of the CD8+brightCD57+ subpopulation. Extent and duration of CD4+ depletion correlated with the cumulative ATG dose (r=0.7, P<0.05, Spearman rank correlation test). CONCLUSION: Therapy with polyclonal antibody ATG induces dose-dependent long-term changes in T-cell lymphocyte subsets, which persist over a period of years.

Adult↗

Chemotherapy-induced differential changes in lymphocyte subsets and natural-killer-cell function in patients with advanced breast cancer.

To elaborate a rational approach to chemoimmunotherapy in humans, information is required as to how current cytotoxic chemotherapy regimens modulate patients' endogenous immune cells. We have studied a group of 16 advanced breast cancer patients who received cyclical cytotoxic chemotherapy (CMF-cyclophosphamide, methotrexate and 5-fluorouracil) and have documented the progressive differential effects of chemotherapy on endogenous immune cells as judged by changes in immunophenotype and absolute numbers of lymphocyte subsets, together with analysis of natural-killer-cell function. Cells with the immunophenotype of natural killer cells and lymphokine-activated killer cells (NK/LAK cells) were well retained, but their function was suboptimal. Additionally, CD8 T cells were well preserved, but the numbers of CD4 T cells decreased with succeeding cycles of chemotherapy; B-cell numbers decreased rapidly from the first cycle of chemotherapy. These cellular changes in humans indicate defined and precisely timed windows of opportunity for introducing in vivo, simple and direct immune stimulation of the cells modulated by chemotherapy, with the possibility of improving therapy and survival in this disease.

Antineoplastic Combined Chemotherapy Protocols↗

Immunogold staining: an alternative method for lymphocyte subset enumeration. Comparison with immunofluorescence microscopy and flow cytometry.

An immunogold staining procedure for light microscopic enumeration of peripheral blood lymphocyte subsets defined by monoclonal antibodies (OKT3, OKT4, OKT8, OKIa1, Leu 1, Leu 4, Leu 2a, Leu 3a, Leu 10, Leu 12, B1) is described. It uses colloidal gold-labeled goat anti-mouse Ig (GAM G40 and GAM G30) as second layer and a methyl-green pyronin counterstain. Performed on small volumes of blood without sophisticated laboratory equipment, this method allows accurate cell type recognition and permanent records, essential for longitudinal observations. By enumerating the gold particles on positively labeled cells, it was shown that the staining reactivity depended on the monoclonal antibody used. Lymphocytes reacting with OKT8 or OKIa1 or B1 exhibited the strongest labeling whereas OKT4+ cells were weakly labeled. When compared with flow cytometry analysis in healthy subjects, the accuracy, precision and sensitivity of both methods were very similar. Similarly, a close correlation (97%) was found between immunogold staining and immunofluorescence microscopy in 35 patients with various diseases suggesting that immunogold staining may be useful in a clinical context.

Antibodies, Monoclonal↗